Thermophile

Can J Microbiol, 1980 Apr, 26(4), 556 - 9
The atypical cell wall composition of Thermomicrobium roseum; Merkel GJ et al.; The cell wall of Thermomicrobium roseum, a Gram-negative, obligately thermophilic bacterium, has a composition unlike other Gram-negative thermophilic organisms . The purified cell wall was composed predominantly of a protein with a monomeric molecular weight of 75 000 . The amino acid composition of this protein revealed high concentrations of proline, glutamic acid, glycine, and alanine.

Appl Environ Microbiol, 1980 Apr, 39(4), 840 - 4
Microbial prevalence in domestic humidifiers; Burge HA et al.; The prevalence of viable thermophilic bacteria and actinomycetes and mesophilic fungi was examined in 145 samples from 110 domestic humidifiers . A total of 72 and 43% of furnace and console humidifier samples, respectively, contained viable thermophilic bacteria, whereas 60 and 72% of these samples produced mesophilic fungal growth . Thermophilic actinomycetes were recovered from seven humidifier samples . Efforts to detect thermophilic actinomycete antigens in 15 humidifier fluid samples were not successful . Antifoulants added to humidifier fluid reservoirs had no apparent effect on microbial frequency . Airborne microbial recoveries did not reflect patterns of humidifier contamination with respect to either kinds or numbers of microorganisms in 20 homes in which volumetric air samples were obtained during humidifier operation.

Proc Natl Acad Sci U S A, 1980 Apr, 77(4), 1922 - 6
Thermally induced biosynthesis of 2'-O-methylguanosine in tRNA from an extreme thermophile, Thermus thermophilus HB27; Kumagai I et al.; The contents of 2'-O-methylguanosine and 1-methyladenosine in unfractionated tRNA obtained from Thermus thermophilus HB27 were found to increase significantly when the bacterium was grown at a higher temperature (80 degrees C) . S-Adenosyl-L-methionine-dependent tRNA (guanosine-2')-methyltransferase (EC 2.1.1.34) and tRNA (adenine-1)-methyltransferase (EC 2.1.1.36) were detected in a cell-free extract of the thermophile, and both of them were partially purified . tRNA (guanosine-2')-methyltransferase specifically catalyzed the methylation of the guanylate residue at position 19 from the 5' end of Escherichia coli tRNAMetf . The amounts of these methyltransferases in the cells and their thermal characteristics seemed to be independent of the growth temperature of the bacterial cells from which the enzymes were extracted . It was inferred that the temperature dependence of the methylation process in vivo is accounted for, not by temperature dependence of enzyme formation, but by that of the enzyme activity.

Mycopathologia, 1980 Mar 31, 70(3), 169 - 79
Thermophillic and thermotolerant fungi isolated from the thermal effluent of nuclear power generating reactors: dispersal of human opportunistic and veterinary pathogenic fungi; Rippon JW et al.; Over a period of a year, samples of water, foam, microbial mat, soil and air were obtained from areas associated with the cooling canal of a nuclear power station . The seventeen sample sites included water in the cooling canal that was thermally enriched and soil and water adjacent to, upstream, downstream and at a distance from the generator . Air samples were taken at the plant and at various distances from the plant . Fifty-two species of thermotolerant and thermophilic fungi were isolated . Of these, eleven species are grouped as opportunistic Mucorales or opportunistic Aspergillus sp . One veterinary pathogen was also isolated (Dactylaria gallopava) . The opportunistic/pathogenic fungi were found primarily in the intake bay, the discharge bay and the cooling canal . Smaller numbers were obtained at both upstream and downstream locations . Soil samples near the cooling canal reflected an enrichment of thermophilous organisms, the previously mentioned opportunistic Mucorales and Aspergillus spp . Their numbers were found to be greater than that usually encountered in a mesophilic environment . However, air and soil samples taken at various distances from the power station indicated no greater abundance of these thermophilous fungi than would be expected from a thermal enriched environment . Our results indicate that there was no significant dissemination of thermophilous fungi from the thermal enriched effluents to the adjacent environment . These findings are consistent with the results of other investigators.

Biochim Biophys Acta, 1980 Mar 7, 590(1), 24 - 33
Isolation and characterization of two rubredoxins from Clostridium thermoaceticum; Yang SS et al.; Two rubredoxins with similar molecular weights (about 6000) have been purified from Clostridium thermoaceticum, a thermophile and strict anaerobe . They exhibit minor differences in several properties like elution pattern from DEAE-cellulose column, isoelectric point, amino acid composition, absorption and EPR spectra and redox potential . Their chemical and physical properties are similar to those of other rubredoxins from anaerobic microorganisms.

Can J Microbiol, 1980 Mar, 26(3), 377 - 84
Purification and properties of polygalacturonic acid trans-eliminase from Bacillus stearothermophilus; Karbassi A et al.; A strain of thermophilic bacteria, Bacillus stearothermophilus, with pectolytic activity has been isolated . It produced an endo-polygalacturonic acid trans-eliminase (endo-PATE, EC 4.2.2.2) extracellularly when grown at 65 degrees C on a pectic acid medium . The PATE was purified 62-fold by the rapid affinity chromatographic method on a Sepharose-polygalacturonamide linked matrix . The absorbed PATE was eluted from the column with a continuous gradient of 0-10(-3)M ethylenediaminetetraacetic acid (EDTA) in phosphate buffer at pH 7.6 . The endo-PATE of this organism was much more heat stable than similar enzymes from the mesophilic Bacillus polymyxa and the thermotolerant Bacillus pumilus . The maximum activity of the enzyme occurred at 70 degrees C . With pectic acid as the substrate, the endo-PATE had an optimal pH of 9.0, the highest optimal pH compared with those of similar enzymes from other species of the genus . The molecular weight of the endo-PATE, as determined by chromatography on a Sephadex G-100 gel column, was 24 000.

J Gen Microbiol, 1980 Mar, 117(1), 201 - 10
Phenotypic and genotypic characterization of some thermophilic species of Bacillus; Sharp RJ et al.; Some thermophilic species of Bacillus were characterized using biochemical tests, antibiotic sensitivity, bacteriocin and bacteriophage sensitivity, esterase patterns, DNA hybridization and % G+C content . The three caldo-active strains of bacilli isolated by Heinen (1971) were compared with strains of B . stearothermophilus . Eight of the strains examined showed characteristics which enabled them to be placed in the three main taxonomic groups suggested by Walker & Wolf (1971) . Two strains showed characteristics of both groups 1 and 3 . Examination of genotype data showed taxonomic groupings which differed from those based on phenotypic characterization.

Am J Public Health, 1980 Mar, 70(3), 241 - 5
Respiratory symptoms and lung function in a sample of Vermont dairymen and industrial workers; Babbott FL Jr et al.; This study reviews the respiratory status of a sample of Vermont male dairy farmers, and a comparison group from industry, matched for age, sex and smoking . Survey instruments included a standardized questionnaire and simple pulmonary function tests . In general, past and present smokers had more respiratory symptoms than never-smokers; and farmers, in all smoking categories, reported symptoms with greater frequency than did their counterparts from industry . Forced vital capacity (FVC) tended to be lower among men with a history of smoking but, within each smoking category, dairymen and factory workers had very similar FVCs . Farmers who had never smoked or who were current cigarette users had lower FEV1/FVC (forced expiratory volume at one second/forced vital capacity) ratios than their controls from industry . Sixteen diarymen demonstrated precipitins to either Micropolyspora faeni (13) or Thermoactinomyces vulgaris, (3), but only one reported a constellation of symptoms compatible with farmer's lung disease . The estimated prevalence of antibodies to thermophilic actinomyces in this farm population was approximately 10 per cent . Although sample sizes were limited, dairymen from small farms tended to be older, have more respiratory symptoms, less satisfactory pulmonary function, and more serologic evidence of exposure to farmer's lung antigens than their counterparts from large farms.

J Biochem (Tokyo), 1980 Mar, 87(3), 745 - 51
Bacillus thermoglucosidius alpha-glucosidase . Temperature dependence of activity and stability; Suzuki Y et al.; A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from an obligate thermophile, Bacillus thermoglucosidius KP 1006, gave a triphasic relationship at pH 6.8 in the van't Hoff plot of Km, in the Arrhenius plot of the first order rate constant of inactivation with 0.1% sodium dodecyl sulfate, and in the logarithmic plot of the maximal fluorescence intensity at 346 nm versus reciprocal of temperature . The respective plots exhibited two breaks at 40 and 61 degrees C, 43 and 62 degrees C, and 40 and 61 degrees C . However, the Arrhenius plot of the molecular activity at pH 6.8 had a single discontinuity at 64 degrees C . These findings, together with thermodynamic quantities for the enzyme, suggest that the thermal conformation changes in the enzyme protein occur around 40--43 degrees C and 61--64 degrees C . The Arrhenius plot of the rate constant of heat inactivation at pH 6.8 was bent at 73 degrees C . Thermodynamic data indicate that the enzyme is transformed from a heat stable form into a heat unstable form at 73 degrees C with temperature elevation . The critical points localized near the minimum, optimum, and maximum temperatures (40, 60, and 72 degrees C) of the cell growth, respectively.

Appl Environ Microbiol, 1980 Mar, 39(3), 584 - 7
Effect of temperature on mineralization by heterotrophic bacteria; Tison DL et al.; When pure cultures of the bacteria Pseudomonas fluorescens (a psychrotroph), Escherichia coli (a mesophile), and SRL 261 (a thermophile) were shifted away from temperatures to which they were adapted, the percentage of substrate mineralized increased (percent mineralized = {substrate respired to CO2}/substrate respired to CO2 + substrate incorporated into biomass} X 100) . The increase in the percent mineralized was larger for larger temperature shifts . Similar responses were observed when natural heterotrophic bacterial populations from sediments of Lake George, N.Y., and a thermophilic algal-bacterial mat community at the Savannah River Plant, Aiken, S.C., were subjected to temperature shifts . These results suggest that an increase in the percent mineralized may be an indication of thermal stress in bacterial populations.

Mikrobiologiia, 1980 Mar-Apr, 49(2), 197 - 201
{Effect of temperature and pH on the ribulose-1,5-diphosphate carboxylase activity of the thermophilic hydrogen bacterium, Pseudomonas thermophila}; Romanova AK et al.; The activity of ribulose 1,5-diphosphate (RDP) carboxylase was found in the soluble fraction of the cytoplasm from sonicated Pseudomonas thermophila K-2 cells . The enzyme is relatively thermolabile and completely loses its activity at 80 degrees C . The activity of RDP carboxylase at 60 degrees C increases by 40% during the first 10 min of heating in the presence of Mg2+ ions, bicarbonate and dithiothreitol, and again decreases if the enzyme is heated over 20 min . The optimum temperature of the enzyme is 50--55 degrees C . The specific activity of the enzyme in fresh preparations under these conditions reaches 0.22 unit per 1 mg of protein in the extract . The calculated value of the activation energy for RDP carboxylase is 6.4 kcal-mole-1, but 11.6 kcal-mole-1 in frozen preparations . The optimal pH is 7.0--7.3 depending on the buffer . The temperature optimum for the enzyme action does not depend on pH within the range of 7.3 to 8.8 . Therefore, RDP carboxylase of Ps . thermophila K-2 differs from RDP carboxylases of mesophilic cultures studied earlier by a higher susceptibility to a decrease in temperature (the enzyme activity is negligible at 30 degrees C), by a lower value of the activation energy at suboptimal temperatures, and by a lower pH optimum of the enzyme action.

Differentiation, 1980 Feb, 16(1), 61 - 9
Perturbance analysis of nuclear determination in Tetrahymena: analysis of mating type frequency variations with reference to binary-switch models; Portnoy S et al.; To test a 3-switch binary element model of mating type determination all possible assignments of the seven mating types of Tetrahymena thermophila to compound switch states have been examined, using a set of 77 experimentally obtained frequency patterns as a base . None of the assignments gives a satisfactory agreement with all the data . The result is similar whether the eighth switch state is considered forbidden or redundant . Moreover, further explorations of models in which the switches are not independent (as originally proposed), but coupled also fail to reveal a satisfactory agreement . Finally some 4-switch models are examined, again without discovering a satisfactory fit, though some evidence of structural relationship among the mating types is provided . The system is more complex than was assumed by the original models . To carry out this analysis certain statistical problems of a more general interest required solution . These include the management of samples from multinomial populations and the provision of efficient estimators for models of this sort.

Differentiation, 1980 Feb, 16(1), 49 - 60
Perturbance analysis of nuclear determination in Tetrahymena: effects of nutrition, cell extracts, and CaCl2 on A/B hybrids; Nanney DL et al.; Mating type frequencies were ascertained among the progeny of crosses of strains A x B, Tetrahymena thermophila under a number of different circumstances . The frequencies are different if the parents are severely starved than if they are well-fed at the time of conjugation; severe starvation of the progeny before the first post-zygotic division has an effect similar to that of starving the parents . Mating type frequencies may also be modified by isolating conjugating pairs into cell extracts before the new macronuclei begin to develop; the changes do not appear to be related in a meaningful way to the mating type of the cells used as a source of the cell extracts . A third means of changing the mating type frequencies involves the exposure of conjugating pairs to CaCl2 solutions . Finally, changed frequency patterns may appear "spontaneously", and reflect either some as yet unsuspected environmental variable, or else an intrinsic metastable state that conditions the probabilities of mating type fixation . With the exception of the starvation effects, the pattern variations seem to fall into two groups . No satisfactory mechanism to account for these results is yet available.

Cell, 1980 Feb, 19(2), 331 - 8
In vitro splicing of the ribosomal RNA precursor in nuclei of Tetrahymena; Zaug AJ et al.; The macronuclear rRNA genes of Tetrahymena thermophila contain a 0.4 kb intervening sequence . In this paper we present evidence that the excision of the intervening sequence from pre-rRNA occurs in vitro in isolated T . thermophila nuclei . The transcription-processing system includes alpha-amanitin to inhibit non-rRNA synthesis and aurintricarboxylic acid to inhibit endogenous nucleases . A discrete 0.4 kb RNA comprises up to 6% of the RNA synthesized in this system . Southern hybridization with restriction fragments of the rDNA (rRNA genes) shows that the 0.4 kb RNA contains the intervening sequence . The size of the 0.4 kb RNA, 410-425 nucleotides, is the size predicted if the entire intervening sequence were excised as a single linear molecule . The 0.4 kb RNA accumulates post-transcriptionally and appears to be stable in vitro . T . pigmentosa strain 6UM, whose rDNA has an intervening sequence of the same size and location as that of T . thermophila, also produces a 0.4 kb RNA in vitro; T . pigmentosa strain 8ALP, with not rDNA intervening sequence, produces no such RNA . The measurement of the accumulation of the excised intervening sequence is a convenient assay for the pre-rRNA splicing activity and a means for characterizing some of the details of the reaction . The resistance of the splicing acticity to concentrations of aurintricarboxylic acid that inhibit other endogenous nucleases should be useful in assaying the splicing enzyme during purification.

Nucleic Acids Res, 1980 Jan 11, 8(1), 43 - 56
A spite specific endonuclease from thermus thermophilus 111, Tth111I; Shinomiya T et al.; A site specific endonuclease with novel specificity has been isolated from Thermus thermophilus strain 111 and named Tth111I . Tth111I cleaves lambda DNA into three fragments of 23.5, 25.7 and 50.8% of the total length, and ColE1 DNA into two fragments of nearly equal length . The sequences around Tth111I cleavage sites of ColE1 and lambda DNA were determined by the Maxam and Gilbert method and the two dimensional mapping method . The results suggest that Tth111I recognizes the DNA sequence (formula: see text) and cleaves the site as indicated by the arrows . Assuming that the first T.A pair in the sequence can be replaced for any base pair, the Tth111I recognition sequence has the symmetry with the two-fold axis as most type II restriction endonucleases do.

Nutr Metab, 1980, 24(2), 65 - 75
{Bacteriological aspects of holding freshly prepared spinach at warm temperatures . Relation between bacterial count and nitrate reduction}; Bomar MT et al.; In sterilized spinach which was inoculated with bacteria, a reduction of NO3- to NO2-, due to their metabolic activity, was detectable, depending on the holding conditions . The reaction rate was highest in the presence of thermophilic bacteria . A lower reaction rate was altogether found with mesophilic bacteria which, depending on counts and holding time, may temporarily cause high nitrite contents . The reaction rate was lowest with cold-tolerant bacteria.

Mikrobiologiia, 1980 Jan-Feb, 49(1), 74 - 7
{Thermostability and ultrastructure of submerged and aerial spores of Actinomyces chrysomallus}; Kuimova TF; Submerged and aerial spores of Actinomyces chrysomallus, strain 2703, producing the antitumour antibiotic chrysomallin occupy, in their thermoresistance, an intermediate position between spores of other mesophilic cultures and endospores of some thermophilic actinomycetes . These spores were heated in water at 90 and 95 degrees C for 10 min . The fraction of highly thermoresistant spores constituted less than 0.001% of all the spores . It was found for the first time in the submerged culture . The structure of the submerged and aerial spores was studied by electron microscopy and was shown to be similar to that of the exotype spores . Heating at high temperatures of the spores often resulted in the strong variability of the culture and in the appearance of proactinomycete-like variants.

Chromosoma, 1980, 78(3), 313 - 25
Chromatin elimination and the genetic organisation of the macronucleus in Tetrahymena thermophila; Cleffmann G; In exponentially growing Tretrahymena thermophila the DNA content of the following structures was determined by cytophotometry: macronuclei of sister cells immediately after division; micronuclei; extranuclear chromatin in dividing cells and postdividers . Further, the development of macronuclear DNA amount in successive cell generations was determined . It was found that chromatin elimination is a frequent process reducing DNA content by about 4% per fission . This chromatin disappears within 20 min after division . The quantity of DNA extruded is highly variable and is different from the micronuclear DNA amount of multiples of it . The frequency of generations with two replication rounds as well as those without replication is estimated to be in the range of 2% each . These findings together with the qualitative difference between micro- and macronuclear DNAs suggest that the macronucleus of Tetrahymena is not entirely composed of complete genomes and that parts of the genetic material must be treated specifically for different sequences either during extrusion or during replication.

Zentralbl Bakteriol Naturwiss, 1980, 135(1), 12 - 21
The effect of thermophilic actinomycetes isolated from compost and animal manure on some strains of Salmonella and Shigella; Makawi AA; Several samples of compost and animal manure were taken from different localities . Eleven isolates of thermophilic actinomycetes were obtained . The morphological, cultural, and physiological properties as well as temperature relation of these isolates were carried out in some detail in order to classify and identify them . The 11 isolates were identified as follows: Thermoactinomyces vulgaris (2 strains), Streptomyces thermofuscus (5 strains) and Thermoactinomyces sacchari (3 strains) . The antibacterial effect of these actinomycetes on the growth of some strains of Salmonella and Shigella was tested . Some of the Streptomyces had an effect on Salmonella . Thermoactinomyces vulgaris had no effect . Shigella strains were not affected.

Folia Parasitol (Praha), 1980, 27(1), 63 - 9
A comparison of developmental dynamics of Dermacentor reticulatus (Fabr.) of different geographic origins and their affection by different microclimate; Daniel M et al.; Developmental dynamics of D . reticulatus ticks of different geographic origins (NE Poland and SW Slovakia) were investigated and compared . A long-term field experiment was conducted in South Moravia in the zone of thermophilic oak forest . Ticks were studied in two different biotopes: in an open grassy area and in deciduous forest . Differences between the tick populations collected in Poland and Slovakia were observed in the duration of developmental cycle and in the outcome of metamorphosis, depending on biotope . The tick development in the open grassy area was more rapid, but with greater losses . The total production of adults was higher in the population originating from Slovakia.

Biochim Biophys Acta, 1980, 595(1), 140 - 5
Modulation of membrane fluidity in a fatty acid auxotrophe of Tetrahymena thermophila; Hill RJ; A mutant of Tetrahymena thermophila has been isolated which requires an unsaturated fatty acid for growth . Pellicles isolated from cultures supplemented with palmitoleic, elaidic, oleic, linoleic and gamma-linolenic acids show widely differing membrane fluidities, as measured by the polarisation of fluorescence technique . In contrast to the behaviour of the wild type organism, the changes in fluidity of the membrane, once induced by supplementation, are permanent . This mutant should prove extremely useful for studying structure-function relationships in the various membrane systems of Tetrahymena.

Cell Motil, 1980, 1(1), 41 - 61
An analysis of spindle ultrastructure during anaphase of micronuclear division in Tetrahymena; LaFountain JR Jr et al.; Mitotic micronuclei were isolated from Tetrahymena thermophila and data on spindle ultrastructure were obtained from serial, transverse sections . Comparison of data from nuclei at meta- and early anaphase with data from nuclei at late anaphase showed that during anaphase, sister kinetochores move from the equator to the spindle poles, but kinetochore translocation occurs without any apparent change in either the number or length of kinetochore microtubules . This unprecedented result is ascribed significance with regard to the mechanism of kinetochore transport since there are only a limited number of ways that result could be achieved . The organization of the peripheral sheath changes during anaphase as evidenced by gaps in the sheath at late anaphase . Numerous kinetochore and non-kinetochore microtubules are located in polar regions of the spindle at late anaphase, whereas those regions contained only peripherally arranged microtubules at earlier stages . Tracking of individual kinetochore microtubules in late anaphase nuclei showed that some of them appeared to become incorporated into the peripheral sheath near the pole . At early and late anaphase, crossbridges connect adjacent microtubules throughout the spindle poleward to the kinetochores, as well as in the interzone.

Z Allg Mikrobiol, 1980, 20(8), 513 - 6
Thermal resistance and viability of asexual spores of thermophilic fungi from composts; Ogundero VW et al.; The growth-temperature relations and thermal tolerance of the asexual spores of thermophilic fungi commonly found in composting plant materials were determined . The optimum temperature range for germination of the spores was 40 degrees - 50 degrees C . The spores of Humicola lanuginosa (GRIFFON and MAUBLANC) BUNCE were able to survive 60 min exposure to 68 degrees C while those of Mucor pusillus COONEY and EMERSON survived only 5 min exposure to 68 degrees C . A determination of the thermal death rates of the spores at 65 degrees C showed a rapid loss of viability of those of Torula thermophila COONEY and EMERSON . The thermal tolerance of the propagules of thermophilic fungi in relation to their special ecological niche is discussed.

Zentralbl Bakteriol Naturwiss, 1980, 135(4), 275 - 89
{Physiological studies on thermophilic hydrocarbon-utilizing hyphae fungi (author's transl)}; Bemmann W et al.; Out of different substrates, thermophilic n-alkane-assimilating hyphae fungi were isolated and the strains of Aspergillus fumigatus and Mucor lusitanicus selected for physiological investigations . The growth curves, obtained in hydrocarbon and glucose at different temperatures, showed three phases: 1) initial growth phase, 2) linear growth phase, and 3) lysis phase . Maximum dry matter and maximum protein content occurred within a range of 35-40 degrees C . During the entire time of growth the DNA content per mg dry matter was almost the same, i.e., 0.1-0.2 per cent . The ascertained specific protease activity of Aspergillus fumigatus correlated with protein synthesis and protein decomposition of the biomass . The results of our investigations are being discussed.

J Immunol Methods, 1980, 34(2), 167 - 75
Detection of proteinases in electrophoretograms of complex mixtures; Westergaard JL et al.; A simple, sensitive technique for detecting proteolytic enzyme zones on electrophoretograms by making contact print zymograms is described . The method is applicable to electrophoretograms prepared on a variety of support media, immunoelectrophoretograms or isoelectric focusing patterns on various media . The contact print zymograms are prepared by placing unfixed, unstained electrophoretograms in contact with a thin film of casein which has diffused into a layer of agarose supported by a hydrophilic polyester film . After staining the casein film with Coomassie blue, the proteolytic zones are detected as clear zones against a blue background . The method can detect as little as 9 ng of trypsin . The utility of the method is illustrated by detection of the proteinase enzymes in thermophilic actinomycete antigen preparations separated by polyacrylamide electrophoresis, crossed immunoelectrophoresis and isoelectric focusing on agarose and granulated dextran supports.

Z Allg Mikrobiol, 1980, 20(2), 105 - 12
{A new selection method for obtaining mutants of Streptomyces glaucoachromogenes producing lambdamycin using the thermophilic phage-host system Tal/Thermoactinomyces vulgaris}; Fleck WF; In order to obtain high-producing mutants of the lambdamycin producer Streptomyces glaucoachromogenes IMET 31 118, a new selection technique using the thermophilic phage-host system Tal/Thermoactinomyces vulgaris, was developed . Lambdamycin is a yellow-green pigment antibiotic of the chromoglycoside type with antimicrobial, antiviral, cancerostatic, and ergotropic activity in vitro and in vivo . The physicochemical properties of lambdamycin resemble those of chartreusin . By means of the selection technique developed the determination of the concentration-dependent influence of lambdamycin on bacterial growth inhibition and Tal-phage development in the agar diffusion sphere of mutagen-treated colonies is possible without time and material-consuming pre-tests . A positive correlation exists between the biosynthetic capacity of mutants on solid media and in fermentation liquors . Using this new selection technique, the titres of lambdamycin in fermentation liquors of mutants could be increased by 10 to 20 fold in comparison to the wild type strain.

J Biochem (Tokyo), 1980 Jan, 87(1), 1 - 13
Purification and thermal stability of several amino acid-specific tRNAs from an extreme thermophile, Thermus thermophilus HB8; Watanabe K et al.; Three species of methionine tRNAs and phenylalanine, tyrosine, and isoleucine tRNAs were purified from an extreme thermophile, Thermus thermophilus HB8 . Formylation studies of the three methionine tRNAs and their codon-specific binding activities to ribosomes showed that two of them (named tRNAf1Met and tRNAf2Met) were initiator tRNAs and the other (named tRNAmMet) was a non-initiator . The tRNAs from T . thermophilus all had melting temperatures of up to ten degrees higher than the corresponding species from E . coli . Most of the species also had slightly higher G+C contents than the corresponding species of E . coli, and each of them contained one mol each of the modified nucleosides, O2'-methylguanosine (Gm), 2-thioribothymidine (s2T), and 1-methyladenosine (m1A) . Their high melting temperatures could be explained by their high G+C contents and the presence of the modified nucleosides, espically s2T . Comparison of the melting temperatures of T . thermophilus tRNAf2Met with those of E . coli tRNAfMet and tRNAmMet at different magnesium concentrations showed that magnesium was also a factor in the thermostability of the thermophile tRNA.

Mol Gen Genet, 1980, 180(1), 129 - 34
Galactokinase-deficient mutants of Tetrahymena thermophila: selection and characterization; Roberts CT Jr et al.; We have isolated a series of mutants of Tetrahymena thermophila which are resistant to inhibition of growth by the galactose analog, 2-deoxygalactose . These mutants were obtained after mutagenesis with nitrosoguanidine and the induction of cytogamy to permit the recovery of recessive mutations induced in the germline micronucleus . Resistance to 2-deoxygalactose is correlated with a decreased rate of growth in galactose minimal medium and greatly reduced levels of galactokinase . The resistant phenotype of the mutants is apparently due to the galactokinase deficiency, which prevents the accumulation of toxic phosphorylated metabolites of 2-deoxygalactose . Genetic analyses reveal that the 2-deoxygalactose resistance alleles segregate as single Mendelian loci . The galactokinase-deficient strains described here represent the first mutants in this organism for which the biochemical basis of the mutant phenotype is known . These mutants, as well as others isolated similarly, should be of value in the elucidation of the mechanisms governing galactokinase gene regulation and in improving techniques of selection for other recessive mutations in Tetrahymena.

Biochim Biophys Acta, 1980, 613(1), 1 - 9
Physicochemical and catalytic properties of thermostable malate dehydrogenase from an extreme thermophile Thermus flavus AT-62; Iijima S et al.; Physicochemical and catalytic properties of thermostable malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37), isolated from an extreme thermophile, Thermus flavus AT-62, were studied . The enzyme had a molecular weight of 67,000 and consisted of two subunits with an identical molecular weight . The helical content of the enzyme was estimated to be about 25% from the circular dichroism spectrum . The amino acid composition of the thermophilic enzyme was similar to that of its mesophilic counterparts . Titration with 5,5'-dithiobis(2-nitrobenzoic acid) showed that the enzyme contained only one sulfhydryl group per subunit . Substrate inhibition by oxaloacetate was observed . The inhibition decreased with increasing temperature, but was still significant at 60 degrees C . The enzyme was remarkably heat stable, without losing activity after incubation at 90 degrees C for 60 min . The melting temperature of the secondary structure of the enzyme was 96 degrees C.

Eur J Biochem, 1980, 107(1), 57 - 65
Amino acid sequence of the proteolipid subunit of the proton-translocating ATPase complex from the thermophilic bacterium PS-3; Hoppe J et al.; The proteolipid subunit of the ATPase complex was identified in whole membranes of the thermophilic bacterium PS-3 by means of a covalent modification with the 14C-labelled inhibitor dicyclohexylcarbodiimide . The proteolipid could be purified from the membrane in free and carbodiimide-modified form by extraction with chloroform/methanol and subsequent carboxymethylcellulose chromatography in mixtures of chloroform/methanol/water . The complete amino acid sequence of the 72-residue polypeptide could be determined by automated solid-phase Edman degradation of the whole protein, and of fragments obtained after cleavage with cyanogen bromide and N-bromosuccinimide . Chemical cleavages and separations of the resulting fragments by gel chromatography were performed in 80% formic acid . The amino acid sequence shows a concentration of hydrophobic amino acids in two segments of about 25 residues at the amino-terminal and carboxy-terminal ends . The polar residues are clustered in the middle of the polypeptide chain . The bound {14C}dicyclohexylcarbodiimide label is recovered exclusively at position 56, which is occupied by a glutamyl residue . The proteolipid from PS-3 exhibits homology to the corresponding ATPase subunit from mitochondria . The carbodiimide-reactive glutamyl residue occurs at the position as in the mitochondrial proteins.

Mikrobiologiia, 1980 Jan-Feb, 49(1), 151 - 5
{In vivo study of the growth characteristics of the microflora of Armenian yogurt}; Erzinkian LA et al.; The microflora from the Armenian yoghurt was studied during its growth on the optimal for the medium (skim milk) using objectives of the Peshkoff system . It was shown that certain organisms of Lactobacterium jogorti tended to branch at the early stages of their growth, and the cells elongated and formed complex coils and helices by the third-fourth day of growth . Compact aggregates of cocci having angular outlines appeared in the culture of Streptococcus thermophilus by the end of the second day of growth . Lysis of individual organisms, and occasionally zones of lysis, were found in the both cultures at certain stages of their growth . Cultures grown for 4-5 days were characterized by wide polymorphism.

Nucleic Acids Symp Ser, 1980, (8), s181 - 4
A second site specific endonuclease from Thermus thermophilus 111,Tth111II; Shinomiya T et al.; A second site specific endonuclease with a novel specificity has been isolated from Thermus thermophilus strain 111 and named Tth111II . The enzyme is active at temperature up to 80 degrees C and requires Mg2+ or Mn2+ for activity . Tth111II cleaves phi X174RFDNA into 11 fragments . From the analysis of 5' terminal sequences of the phi X174RFDNA fragments produced by Tth111II action, it was concluded that Tth111II recognized the DNA sequence (See formula in text) and cleaved the sites as indicated by arrows.

Ultrastruct Pathol, 1980 Jan-Mar, 1(1), 19 - 37
Langerhans cells and serum precipitating antibodies against fungal antigens in bronchioloalveolar cell carcinoma: possible association with pulmonary eosinophilic granuloma; Hammar SP et al.; In an ultrastructural study of 37 cases of bronchioloalveolar cell carcinoma (BAC), we identified seven cases (19%) in which Langerhans cells (LC) were closely associated with tumor cells . Serum precipitating antibodies against Aspergillus species and/or thermophilic actinomyces were present in five BAC patients whose tumors contained LC and in six patients whose tumors lacked LC . In a simultaneous study we identified marked atypical bronchiolar and alveolar lining cell hypertrophy and hyperplasma in pulmonary eosinophilic granuloma (PEG) . Our data plus current information suggesting that PEG is a form of hypersensitivity pneumonitis support our hypothesis that those cases of BAC in which LC are present may rise in localized or diffuse pulmonary scars caused by PEG.

Can J Microbiol, 1980 Jan, 26(1), 58 - 63
Effect of prostaglandin E1-induced elevation of cyclic AMP on glucose repression in the lactic streptococci; Ratliff TL et al.; Cyclic adenosine 3', 5' -monophosphate (cAMP) activity was observed in Streptococcus lactis C2, Streptococcus cremoris C10, Streptococcus diacetlactis 18-16, and Streptococcus thermophilus C3 . In vitro assays of cell-free extracts obtaianed from S . lactis C2 showed that the cAMP-associated enzymes adenyl cyclase and phosphodiesterase were also present . In vitro experiments showed that prostaglandin E1 (PGE) stimulation of adenyl cyclase increased cAMP concentrations approximately fivefold, and in vivo studies showed that PGE treatment of S . lactis C2 increased intracellular cAMP concentrations twofold . Futhermore, PGE-induced elevation of intracellular cAMP levels was shown to prevent the repression of ss-D-phosphogalactoside galactohydrolase synthesis by glucose.

Proc Natl Acad Sci U S A, 1980 Jan, 77(1), 147 - 51
Properties of a copper-containing cytochrome c1aa3 complex: a terminal oxidase of the extreme thermophile Thermus thermophilus HB8; Fee JA et al.; From the plasma membrane of Thermus thermophilus HB8 we have partially purified a detergent-solubilized complex of cytochromes a and c1 that actively catalyzes the transfer of electrons from ascorbate via a redox dye to oxygen . The complex is composed of two types of polypeptides, with molecular weights of approximately 55,000 and 33,000 . Quantitative analysis revealed the presence of heme a, heme c, and copper in a ratio of 2:1:2, with the heme a being present at 10 +/- 1.3 nmol/mg of protein . The heme c was shown to be associated with the molecular weight 33,000 peptide and is suggested to be of the c1 type . The optical and electron paramagnetic resonance properties of this complex were found to be similar to those of eukaryotic cytochrome oxidase, suggesting the following arrangement of chromophores: a magnetically isolated cytochrome c1 and an oxygen-reducing functional unit consisting of two heme a groups and two copper ions associated with one or more larger peptides.

Biochemistry, 1979 Dec 11, 18(25), 5698 - 703
Thermal stability and protein structure; Argos P et al.; Amino acid sequences have been compared for thermophilic and mesophilic molecules of ferredoxin, glyceraldehyde-3-phosphate dehydrogenase, and lactate dehydrogenase . It is shown that Gly, Ser, Ser, Lys, and Asp in mesophiles are generally substituted by Ala, Ala, Thr, Arg, and Glu, respectively, in thermophiles . These exchanges suggest that thermal stability can be achieved by the addition of many small changes throughout the molecule without significant change in the backbone conformation . Their overall effect is primarily to increase internal and decrease external hydrophobicity as well as to favor helix stabilizing residues in helices . These substitutions minimize interruption of function or internal residue packing arrangements . Although the analysis has been confined to the above-mentioned molecules, the observed stabilizing principles may be more generally applicable.

Biochemistry, 1979 Dec 11, 18(25), 5693 - 7
Denaturation of thermophilic ferricytochrome c-552 by acid, guanidine hydrochloride, and heat; Hon-nami K et al.; The denaturation of Thermus thermophilus cytochrome c-552 by acid, guanidine hydrochloride, and heat was studied by measuring the changes in absorption and circular dichroism . Cytochrome c-552 was remarkably resistant to acid; the pK of the transition from the low- to the high-spin form was roughly 0.3 . The effect of guanidine hydrochloride on the heme iron-methionine bond of Thermus and horse cytochromes c was also investigated; a comparison of the free-energy changes for the displacement of the bond indicated that the coordination in cytochrome c-552 is highly stable . The spectra of guanidine hydrochloride unfolded cytochrome c-552 were dependent on the pH; the titration curve showed the presence of a cooperative single transition of pK = 4.7, with a one-proton dissociation, suggesting the ionization of a histidine residue . In the presence of guanidine hydrochloride, the influence of the heat on the ligand bond in cytochrome c-552 was studied . The van't Hoff plots of the reaction were biphasic . The enthalpy changes in the higher temperature range were independent on the guanidine hydrochloride concentration, while those in the lower range were not.

J Cell Sci, 1979 Dec, 40, 111 - 23
The regulation of amounts and proportions of genetic elements in the macronuclei of Tetrahymena thermophila strains of diverse karyotype; Seyfert HM et al.; Measurements of the micronuclear DNA content of clones with assumed different degrees of micronuclear ploidy confirmed the triploid nature of one clone . The micronuclear DNA content of a presumptive haploid clone was found to be slightly higher than expected, whereas one of two aneuploid clones had an unexpectedly low micronuclear DNA content . The macronuclear DNA content of cells with macronuclei which had developed from triploid, diploid and probably haploid and aneuploid macronuclear-Anlagen is very similar . Specifically, the smallest individual macronuclear DNA contents are consistently found within the same size class in all clones tested . Cell volumes, RNA and protein contents are alike in all clones tested . Only the growth rate and maximal density reached of one out of two aneuploid clones is reduced in comparison with the very similar other clones . The results are discussed with reference to the regulation of macronuclear DNA content, and to the compensation of gene-dosage.

J Gen Microbiol, 1979 Dec, 115(2), 451 - 6
Mass selection of conditional mating mutants of Tetrahymena thermophila; Wolfe J; The mating reaction in Tetrahymena thermophila includes a starvation period and two distinct cell interactions, co-stimulation and cell pairing, before the cells are cytoplasmically joined as conjugants . A selection procedure for harvesting mutants unable to mate at a restrictive temperature has been developed . A conjugant pair consisting of one cycloheximide-resistant cell and one wild-type cell (cycloheximide-sensitive) was itself sensitive to the drug . By adding cycloheximide and nutrient medium to a cross made at the restrictive and grow . Repetition of the selection procedure enriched for cells unable to conjugate at the restrictive temperature . The selected cells were able to grow at 38 degrees C and could conjugate at 28 degrees C . This procedure may be narrowed to select specifically for cell interaction mutants.

J Cell Physiol, 1979 Dec, 101(3), 503 - 13
Ribosome biosynthesis in Tetrahymena thermophila . IV . Regulation of ribosomal RNA synthesis in growing and growth arrested cells; Sutton CA et al.; Three parameters involved in the production of new ribosomal RNA (rRNA) were measured in Tetrahymena thermophilia: (i) the rate of synthesis of the rRNA precursor, (ii) the rate of processing of the RNA precursor and rRNA intermediates and (iii) the efficiency of utilization of the rRNA precursor in producing mature ribosomal RNA . These parameters were measured in cells in exponential growth and in cells starved in a dilute salt solution . Growing cells synthesize rRNA 20 times faster and process rRNA precursors and intermediates 10 to 15 times more rapidly than do starved cells . Both utilize their rRNA precursors with an efficiency of one in converting them to mature rRNA.

J Biochem (Tokyo), 1979 Dec, 86(6), 1679 - 85
Occurrence of GTP cyclohydrolase I in Bacillus stearothermophilus; Suzuki Y et al.; A GTP cyclohydrolase which catalyzes the removal of carbon 8 of GTP as formic acid to yield a single pteridine compound occurs in an obligate thermophile Bacillus stearothermophilus ATCC 8005 . The enzyme was purified 5.5-fold . Its molecular weight and Stoke's radius were estimated as 105,000 and 45.3 A, respectively . The Km for GTP was 0.98 microM . The temperature and pH optima for activity were 60-65 degrees C and 8.0-8.4, respectively . No divalent cation was required for the reaction . The pteridine product was 3'-triphosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)-7,8-dihydropteridine (dihydroneopterin triphosphate), identified by isolating its immediate derivative, 2',3'-cyclic phosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)pteridine (neopterin cyclic phosphate) . The radioactive product from {8-14C}GTP agreed with 14C-formate . Molar ratio of formate release to pteridine formation was 1.0.

Infect Immun, 1979 Dec, 26(3), 1057 - 64
Crossed immunoelectrophoretic analysis of two antigen extracts of Thermoactinomyces candidus; Hollick GE et al.; A pyridine extract antigen and a double-dialysis antigen (DDA) obtained from Thermoactinomyces candidus were analyzed by crossed immunoelectrophoresis . In addition, the heat lability, pronase sensitivity, and isolectric points of the components of the DDA were determined . By using antisera raised against crude pyridine extract antigen, two immunogenic components were resolved by crossed immunoelectrophoresis . A similar analysis of DDA using antisera raised against crude DDA revealed 15 immunogens . All but six components were heat labile, whereas pronase had little effect on the number of resolvable components . Intermediate gel crossed immunoelectrophoresis using antiserum raised to whole spores detected six immunogenic components, four of which were also detected by the anti-DDA serum . A total of 19 bands were obtained when the DDA was subjected to flatbed isoelectric focusing on polyacrylamide gels . The isoelectric points for the various components were found to range from 3.5 to 5.7 . Crossed immunoelectrophoresis using isoelectric focusing in the first dimension yielded at least 16 immunogenic components . Six components with isoelectric points falling in the range of 4.5 to 6.4 were found to be resistant to heat . A comparison with antigens obtained from other thermophilic actinomycetes is presented.

J Biochem (Tokyo), 1979 Dec, 86(6), 1687 - 95
A double-alpha c-type cytochrome, cytochrome c-555, 549, from an extreme thermophile, Thermus thermophilus HB8; Hon-Nami K; A "double-alpha" c-type cytochrome, cytochrome c-555, 549, was isolated from the membrane fraction of an extreme thermophile, Thermus thermophilus HB8, and highly purified by chromatographies on DEAE-cellulose and Sephadex G-75 and by isoelectric focusing . The absorption maxima were at 554.8, 548.6, 522, and 417 nm in the reduced form, and at 528, 409, and 360 nm in the oxidized form . The double alpha-peak of this cytochrome was enhanced at liquid nitrogen temperature . The cytochrome contained one heme c group per protein molecule . The isoelectric point, midpoint redox potential and molecular weight were pH 4.0, +0.206 V and about 10,000, respectively . Cytochrome c-555, 549 is highly thermostable.

Biophys Struct Mech, 1979 Dec, 6(1), 31 - 7
Thermal stability of soluble mitochondrial H+-ATPase; Kiladze AA et al.; ATPase melting has been studied by circular dichroism and differential scanning microcalorimetry . Decomposition of the alpha-helix of H+-ATPase (in which about 80% of the peptide groups of the enzyme are involved) following thermal treatment is shown to proceed gradually, beginning with room temperature . Effect of nucleotides upon melting is detected in the range of 20 degrees--40 degrees C . Above 40 degrees C, the pattern of thermal decomposition of the three-dimensional structure of H+-ATPase is independent of the nature of nucleotides present . Highly stable alpha-helical sites have been found in the enzyme molecule . Possible mechanism of formation of such sites is discussed, and the results obtained are compared with data on thermal stability of ATPase from thermophilic bacteria . Structural changes in the molecule following thermal treatment are compared with ATPase activity changes under similar experimental conditions.

Biochim Biophys Acta, 1979 Nov 22, 565(1), 154 - 60
Stability of bacterial messenger RNA in mesophiles and thermophiles; Stenesh J et al.; The decay of {3H}uridine-labeled mRNA was measured in the mesophile, Bacillus licheniformis (grown at 37 degrees C and 46 degrees C), and in the thermophile, Bacillus stearothermophilus (grown at 46 degrees C and 55 degrees C) . For each organism, the half-life of the mRNA decreased as the growth temperature was increased . The stability index (half-life of mRNA/doubling time of cells), however, was remarkably constant for each organism regardless of the growth temperature . It is concluded that these results support the concept that kinetic considerations play a significant role in the explanation of thermophily.

Mikrobiologiia, 1979 Nov-Dec, 48(6), 1113 - 7
{Distribution of thermophilic sulfate reducing bacteria in the oil-bearing strata of Apsheron and Western Siberia}; Rozanova EP et al.; Enrichment cultures of sulfate-reducing bacteria were obtained by inoculating the water of high-temperature gas and oil-bearing strata . A study of the morphology of these cells has shown that the thermophilic bacterium Desulfovibrio thermophilus occurs in the deposits of the Apsheron Peninsula while Desulfootomaculum nigrificans is found in the deposits of West Siberia . The former organism is involved in the accumulation of H2S in the Apsheron strata . The temperature of the strata is believed to be the factor which regulates the incidence of bacteria.

Mech Ageing Dev, 1979 Nov, 11(4), 253 - 68
Germinal aging in Tetrahymena thermophila; Simon EM et al.; The manifestations of germinal aging in the ciliate Tetrahymena thermophila include death of the cells at conjugation and macronuclear retention in which the normal replacement of the old macronucleus by a new one fails to occur . Available data suggest that methods of routine maintenance that reduce the number of fissions may delay aging . Differences in breeding performance following maintenance for 1-5 years in axenic peptone broth vs . bacterized Cerophyl were not significant; those following maintenance at different temperatures were significant . The analysis of several hundred crosses is consistent with a random mutational basis for aging in the micronucleus and does not support the hypothesis of an age-correlated program in which the rate of deterioration increases with time . Following routine cultural maintenance for as long as 9-11 years, some lines show no deficiencies in their ability to produce viable progeny, and sublines of the same clone frequently differ significantly in their breeding performance . Moreover, breeding degeneration occurs at constant, but different, rates in different inbred strains.

J Biochem (Tokyo), 1979 Nov, 86(5), 1427 - 32
Addition of short guanylyl blocks to oligonucleotide primers with a thermophilic polynucleotide phosphorylase . Its application to the synthesis of oligonucleotides containing guanylyl residues; Kikuchi Y et al.; Polynucleotide phosphorylase from Thermus thermophilus catalyzed the addition of short guanylyl blocks from GDP to the 3'-hydroxyl termini of oligonucleotide primers at low temperature in a simple reaction mixture . Polyguanylic acid formation was inhibited at 37 degrees C, but the addition of one or two guanylyl residues to oligonucleotide primers proceeded in high yields . The reaction was applied to the synthesis of oligonucleotides containing guanylyl residues at the 3'-end . Using (Ap)2A and (Up)2U as primers, (Ap)3G, (Ap)3GpG, and (Up)3G were synthesized in yields of 25--52% . (Ap)2GpG was synthesized from ApA and GDP in a yield of 13%.

J Cell Physiol, 1979 Nov, 101(2), 349 - 58
Ribosome biosynthesis in Tetrahymena thermophila . III . Regulation of ribosomal RNA degradation in growing and growth arrested cells; Sutton CA et al.; We have measured the turnover rate of ribosomal RNA in exponentially growing Tetrahymena thermophila cells, cells entering the plateau phase of growth, and nutrient-deprived (starved) cells . Ribosomal RNA is stable in cells in early log phase growth but it begins to turnover as the cells begin a deceleratory growth phase prior to entering a plateau state . Likewise, rRNA in cells transferred from early log phase growth to a starvation medium begins to be degraded immediately upon starvation . In both cases the degradation of rRNA exhibits biphasic kinetics . A rapid initial exponential degradation with a half time of nine and one-half hours lasting for six hours is followed by a slower exponential degradation with a half-life of 35 hours . When starved cells are transferred to fresh growth medium turnover of rRNA ceases . The evidence presented suggests that the alteration in degradation rate is a regulated process which is most likely independent of the cell cycle.

J Bacteriol, 1979 Nov, 140(2), 543 - 6
Protein turnover in the extreme thermophile Thermus aquaticus; Kenkel T et al.; Protein turnover in the extreme bacterial thermophile Thermus aquaticus was examined in exponential cultures at 75 degrees C . The relative amount of {3H}leucine incorporated into trichloroacetic acid-insoluble material was stable in pulse-chase experiments assayed over 2.5 h . The trichloroacetic acid-insoluble radioactive leucine was stable upon the addition of chloramphenicol, which blocks protein synthesis in T . aquaticus . The specific activity of a phosphate-repressible alkaline phosphatase, investigated in the presence of chloramphenicol, did not decrease . The addition of excess orthophosphate to cultures derepressed for the alkaline phosphatase did not show a marked effect on the specific activity over a 2-h period . On the basis of these four experiments, it does not appear that a high protein turnover rate is essential for the thermophily of T . aquaticus at 75 degrees C.

J Gen Microbiol, 1979 Nov, 115(1), 253 - 4
Thermophilic and thermotolerant fungi in poultry droppings in Nigeria; Ogundero VW; Ten species of fungi were obtained from poultry droppings in Nigeria . Six of these are true thermophiles while the other four are thermotolerant . Aspergillus fumigatus Fresenius, Mucor pusillus Lindt and Thermoascus aurantiacus Stolk are known human pathogens . Except for M . pusillus, all the thermotolerant species had a higher occurrence at 45 degrees C while the thermophilic varieties were readily obtained at 50 degrees C.

Eur J Biochem, 1979 Nov, 101(2), 317 - 24
5'-Methylthioadenosine phosphorylase from Caldariella acidophila . Purification and properties; Carteni'-Farina M et al.; The occurrence of 5'-methylthioadenosine phosphorylase in Caldariella acidophila, a thermophilic bacterium growing optimally at 87 degrees C, is reported . It represents the first example in prokaryotes of a phosphoryolytic cleavage of the thioether . The reaction products, purified by ion-exchange chromatography, have been identified as 5-methylthioribose-1-phosphate and adenine by several analytical procedures . The enzyme has been purified to homogeneity in 32% yield by using DEAE-cellulose and hydroxyapatite chromatography, gel filtration and isoelectric focusing . The enzyme shows a high degree of thermophilicity, its temperature optimum being at 93 degrees C; furthermore no loss of activity is observable after exposure for 1 h at 100 degrees C . The kinetic data indicate a sequential mechanism of the reaction . The apparent Km values are 0.095 mM for 5'-methylthioadenosine and 6.1 mM for phosphate . The specificity of the reaction is rather strict . Experiments performed with analogues of the substrate, i.e . 5'-methylthioinosine, 5'-dimethylthioadenosine sulfonium salt, 5'-n-butylthioadenosine, 5'-isobutylthioadenosine, 5'-isobutylthioinosine, adenosylhomocysteine, 5'-thioethanoladenosine, adenosine, indicate the relevance of the adenine amino group and the sulfur in thioether form in the binding to the enzyme protein.

Biochemistry, 1979 Oct 16, 18(21), 4702 - 7
Lipoamide dehydrogenase from Malbranchea pulchella: isolation and characterization; McKay DJ et al.; Lipoamide dehydrogenase (EC 1.6.4.3) has been isolated from a total homogenate of frozen mycelium of the thermophilic fungus Malbranchea pulchella var . sulfurea by a three-step procedure involving ammonium sulfate fractionation, Procion Brilliant Blue M-R--Sepharose 4B chromatography, and hydroxylapatite chromatography . The second step is the key purification step with the Procion Brilliant Blue M-R dye acting as an affinity ligand for the enzyme . The purified enzyme gave a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate . The enzyme is a dimer of molecular weight 102 000, and each monomer of 51 000 molecular weight binds one molecule of flavin adenine dinucleotide . Other properties determined include a pH optimum of 8.2, a strong specificity for the substrates dihydrolipoamide and nicotinamide adenine dinucleotide, the apparent lack of multiple enzymic forms, the presence of diaphorase activity, and resistance to temperature denaturation up to 60 degrees C . The amino acid composition and absorption spectrum of the enzyme were also determined . The properties of lipoamide dehydrogenase from this source are very similar to those reported for the enzyme from serveral other sources.

Biochim Biophys Acta, 1979 Oct 11, 570(2), 406 - 10
Regulatory characteristics of phosphoenolpyruvate carboxylase from the extreme thermophile, Thermus aquaticus; Sundaram TK et al.; Phosphoenolpyruvate carboxylase from the extremely thermophilic bacterium, Thermus aquaticus YT-1, exhibits a virtually absolute requirement for acetyl CoA and there is strong positive cooperativity in the interaction of this activator with the enzyme . Several tricarboxylic acid cycle intermediates inhibit the enzyme . These findings suggest an anaplerotic role for the enzyme and an allosteric modulation of its activity by acetyl CoA and tricarboxylic acid cycle intermediates.

Biochim Biophys Acta, 1979 Oct 10, 548(1), 139 - 46
Exchange integral for a variety of tetranuclear ferredoxins; Blum H et al.; The temperature dependence of EPR spectra of oxidized {4Fe-4S}(-1,-2) ferredoxins (previously designated HiPIP) and a reduced {4Fe-4S}(-2,-3) ferredoxin have been analyzed so as to determine the energy of a low-lying excited electronic state . The values obtained were: Center S-3 from beef heart, 44 cm-1; Center S-3 from mung bean, 53 cm-1; the {4Fe-4S}(-1,-2) ferredoxin from Thermus thermophilus, 78 cm-1; Center N-2 of NADH ubiquinone reductase, 83 cm-1 . Increasing axial distortion in the EPR spectra of the {4Fe-4S}(-1,-2), ferrodoxins was associated with higher energy differences . Center N-2, a {Fe-4S}(-2,-3) iron-sulfur cluster does not fit this relationship.

J Cell Sci, 1979 Oct, 39, 299 - 312
Mitochondrial associations with specific microtubular components of the cortex of Tetrahymena thermophila . I . Cortical patterning of mitochondria; Aufderheide K; Many of the mitochondria of Tetrahymena thermophila are localized in the cell cortex in regular, identifiable patterns . Two different mitochondrial patterns are seen; whether a cell expresses one or the other type apparently depends upon nutrient conditions in the culture and not upon other factors tested . Consistent associations between cortical mitochondria and certain of the cortical microtubular bands are seen at the light-microscopic level . Electron-microscopic observations confirm this and, furthermore, identify ultrastructural associations between cortical microtubules and the cortically located mitochondria . It appears that the cortical microtubular arrays serve as a guide for the localization (and thus patterning) of the cortical mitochondria.

Cell, 1979 Oct, 18(2), 525 - 32
The intervening sequence in the 26S rRNA coding region of T . thermophila is transcribed within the largest stable precursor for rRNA; Din N et al.; We studied the transcription of the intervening sequence in the 26S rRNA coding region of the extrachromosomal rDNA molecules in the macronucleus of T . thermophila by hybridization of purified nuclear rRNA precursors or cytoplasmic 26S rRNA to purified native rDNA or specific rDNA restriction fragments . Examination of R loop hybrids in the electron microscope and analyses of S1-protected rDNA fragments in alkaline agarose gels showed that mature 26S rRNA, nuclear pre-26S rRNA and a fraction of the pre-rRNA molecules containing both the sequences for 17S and 26S rRNA all lack the region corresponding to the intervening sequence . The rest of the pre-rRNA molecules, however, hybridize in a colinear fashion to the whole coding region, and thus must contain the intervening sequence . We can conclude from these results that the intervening sequence is transcribed within the primary transcription product of the rDNA, and that the post-transcriptional removal of the intervening RNA sequence is a very early processing event in the organism.

J Biochem (Tokyo), 1979 Oct, 86(4), 1055 - 65
Kietics of thermal unfolding and refolding of thermostable phosphoglycerate kinase; Nojima H et al.; The kinetics of denaturation by guanidine hydrochloride (GuHCl) of a thermostable phosphoglycerate kinase (PGK) extracted from Thermus thermophilus and of yeast PGK at neutral pH were studied by circular dichroism . Denaturation by GuHCl proceeded as a first-order reaction . The activation free energy of the denaturation reactions (delta Gf not identical to ) in the absence of GuHCl was estimated to be 32.7 kcal/mol for T . thermophilus PGK and 27.9 kcal/mol for yeast PGK (at 25 degrees C) . Measurements of the rate constants at various temperatures indicated that delta Gf not identical to has maximum values at 29 degrees C for T . thermophilus PGK and at 20 degrees C for yeast PGK, and that the temperature dependences of delta Gf not identical to, delta Hf not identical to, and delta Sf not identical to for T . thermophilus PGK are smaller than those of yeast PGK . Values of delta Sf not identical to for thermal denaturation for both PGK's are approximately 200 e.u.

Proc Natl Acad Sci U S A, 1979 Oct, 76(10), 5051 - 5
Localization of transcribed regions on extrachromosomal ribosomal RNA genes of Tetrahymena thermophila by R-loop mapping; Cech TR et al.; R-loop hybridization and electron microscopy were used to map the RNA transcription products of the extrachromosomal rRNA genes of Tetrahymena thermophila . The mature 17S and 26S rRNAs and the nuclear 35S pre-rRNA and pre-26S rRNA were located with a precision of approximately 100 base pairs . A 370-base pair intervening sequence was found in the 26S coding region . It has the same size and relative location as that found in Tetrahymena pigmentosa {Wild, M.A . & Gall, J.G . (1979) Cell 16, 565-573} . One class of R-loop structures formed by nuclear pre-rRNA provided preliminary evidence for a primary transcript that contains the intervening sequence . The results suggested a processing scheme in which splicing of the intervening sequences is followed by a series of strand cleavages to give the mature 17S and 26S rRNAs . Analysis of the data also showed that RNA . DNA and DNA . DNA duplexes, when mounted for electron microscopy by the R-loop procedure, have the same length per base pair within 4%.

Proc Natl Acad Sci U S A, 1979 Oct, 76(10), 4857 - 61
Micronuclei of Tetrahymena contain two types of histone H3; Allis CD et al.; Evidence is presented that micronuclei of Tetrahymena thermophila contain significant amounts of two types of histone H3 . One is indistinguishable from that found in macronuclei and the other is unique to micronuclei . The micronucleus-specific H3 has a slightly faster mobility than the common H3 in three different gel systems (both of these species were artifactually lost during procedures for histone preparation in previous studies) . Both micronuclear H3s appear to contain a single cysteine residue and are present in sucrose gradient-purified nucleosomes . Acid extracts from micronuclei also contain three prominent high molecular weight proteins that also were lost during previous procedures . These proteins are present in extracts from oligomers but are not observed in extracts from mononucleosomes, suggesting that they may be associated with linker regions between nucleosomes.

J Gen Microbiol, 1979 Oct, 114(2), 401 - 8
Isolation and partial characterization of four plasmids from antibiotic-resistant thermophilic bacilli; Bingham AH et al.; Twenty-nine antibiotic-resistant isolates of thermophilic bacilli were examined for the presence of covalently closed circular duplex DNA molecules by agarose-gel electrophoresis and caesium chloride-ethidium bromide density gradient centrifugation . Five of the 29 strains tested contained covalently closed circular molecules . Two of the streptomycin-resistant strains contained the same two plasmids: pAB118A of molecular weight 4.9 X 10(6) (7.0 kilobases) and pAB118B of molecular weight 3.0 X 10(6) (4.3 kilobases) . Two of the tetracycline-resistant strains each contained a plasmid (pAB124) of molecular weight 2.9 X 10(6) (4.14 kilobases), while a third harboured a small plasmid (pAB128) of molecular weight 2.5 X 10(6) (3.57 kilobases) . These plasmids were digested with 19 different restriction endonucleases and the numbers of cleavage sites were determined . Transformation of Bacillus subtilis (168 (Trp-) with purified plasmid DNA indicated that pAB124 conferred tetracycline resistance on the host.

J Biochem (Tokyo), 1979 Oct, 86(4), 893 - 905
Nucleotide sequence of formylmethionine tRNA from an extreme thermophile, Thermus thermophilus HB8; Watanabe K et al.; The nucleotide sequence of formylmethionine tRNA from an extreme thermophile, Thermus thermophilus HB8, was determined by a combination of classical methods using unlabeled samples to determine the sequences of the oligonucleotides of RNase T1 and RNase A digests and a rapid sequencing gel technique using 5'-32P labeled samples to determine overlapping sequences . Formylmethionine tRNA from T . thermophilus is composed of two species, tRNAf1Met and tRNAf2Met . Their nucleotide sequences are almost identical, and are also almost identical with that of E . coli tRNAfMet, except for slight modifications and replacements . Both species have modifications at three points which do not exist in E . coli tRNAfMet: 2'-O-methylation at G19, N-1-methylation at A59 and 2-thiolation at T55 . Moreover U51 in E . coli tRNAfMet is replaced by C51 in both species, so that a G-C pair is formed between this C51 and G65 . tRNAf2Met has a reversed G-C pair at positions 52 and 64 compared with those in tRNAf1Met and E . coli tRNAfMet . Other regions are mostly the same as those in all prokaryotic initiator tRNAs so far reported . The thermostability of these thermophile initiator tRNAs is discussed in relation to their unique modifications.

J Biol Chem, 1979 Sep 25, 254(18), 8720 - 2
A new polyamine, thermospermine, 1,12-diamino-4,8-diazadodecane, from an extreme thermophile; Oshima T; A new polyamine has been extracted from an extreme thermophile, Thermus thermophilus, and its chemical structure was determined as 1,12-diamino-4,8-diazadodecane, NH2(CH2)3NH(CH2)3NH(CH2)4NH2, based on its proton NMR, 13C NMR, and mass spectra . A trivial name "thermospermine" is proposed for the new compound.

Biochim Biophys Acta, 1979 Sep 21, 556(2), 265 - 77
Purification and partial characterization of a procaryotic glycoprotein from the plasma membrane of Thermoplasma acidophilum; Yang LL et al.; The obligate, thermophilic, acidophilic mycoplasma, Thermoplasma acidophilum, grows optimally at 56 degrees C and pH 2.0 . Its plasma membrane possessed 21--22 protein bands that were resolved by polyacrylamide gel electrophoresis . One major membrane protein, molecular weight 152 000, which stained for carbohydrate with periodic acid-Schiff reagent, accounted for 32% (w/w) of the total membrane proteins . It was isolated and further purified by concanavalin A affinity chromatography . The carbohydrate content amounted to less than 10% (w/w) compared to that of the entire glycoprotein . The carbohydrate moiety consisted mainly of mannose residues with branched alpha 1 leads to 2 linkages at the non-reducing ends of the glycopeptide as determined by permethylation followed by gas chromatography-mass spectrometry analysis . The reducing end was an N-glycosidic linkage between asparagine and N-acetylglucosamine . The amino acid composition of this glycoprotein showed 62 mol% hydrophobic residues, while the acidic amino acid content contributed 9 mol% more than that of the basic amino acids . The existence of membrane glycoproteins in the procaryotic, wall-less T . acidophilum may provide a protective coat for the plasma membrane . The stereochemistry and the conformation of the carbohydrate chains, in conjunciton with water turgor, may contribute to the rigidity of the membrane and the cation binding.

J Cell Physiol, 1979 Sep, 100(3), 407 - 11
Peptidase activity in Tetrahymena; Zdanowski MK et al.; This report strongly suggests that two compartments in Tetrahymena thermophila contain peptidase activity: the cytoplasm and the outer cell surface . Determinations of amino acid concentrations in the extracellular medium upon incubation of cells with peptides suggest that the surface-bound peptidase activity hydrolyses di- and tri-phenylalanine equally fast on a molar basis . Growth experiments designed to characterize the in vivo peptidase specificities showed that both T . thermophila and T . pyriformis can use L-leucyl-L-leucine, but not L-leucyl-D-leucine as a leucine donor . These results are independent of whether the cells form food vacuoles or not.

J Bacteriol, 1979 Sep, 139(3), 800 - 10
Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum; Wiegel J et al.; Thirteen strains of a strict anaerobic, extreme thermophilic bacterium were isolated from soil samples of moderate temperature, from a sewage plant in Georgia, and from hot springs in Utah and Wyoming . They were identified as strains of Clostridium thermohydrosulfuricum . The guanosine + cytosine content (moles percent) was 37.6 (determined by buoyant density) and 34.1 (determined by melting temperature) . All strains required a factor present in yeast extract or tryptone growth . Growth characteristics were as follows: a pH range of 5 to 9, with the optimum between 6.9 to 7.5, in a temperature range of 40 to 78 degrees C, with the optimum at 68 degrees C . The doubling time, when grown on glucose at temperature and pH optima, was 1.2 h . The main products of glucose fermentation were ethanol, lactate, acetate, CO2, and H2 . The fermentation was inhibited by H2 . Formation of spores occurred easily on glucose-agar medium or when cultures growing at temperatures above 65 degrees C were allowed to cool to temperature below 55 degrees C . C . thermohydrosulfuricum occurs widely distributed in the natural environment.

Biochemistry, 1979 Aug 21, 18(17), 3647 - 53
Pyruvate carboxylase from a thermophilic Bacillus: some molecular characteristics; Libor S et al.; Analysis of the native enzyme and of the subunits produced upon its denaturation shows that pyruvate carboxylase from a thermophilic Bacillus is a tetramer with a molecular weight (mean value) of 558,000 and that the four polypeptide subunits are probably identical . The three functions (carboxyl carrier, carboxylation, and carboxyl transfer) in the pyruvate carboxylation reaction must therefore reside in this quarter-molecular polypeptide . The enzyme molecule contains four atoms of zinc and four molecules of D-biotin, and in the electron microscope the disposition of its four subunits presents a rhombic appearance . Reaction of the denatured enzyme with 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) reveals 10 sulfhydryl groups/subunit . In the native enzyme less than one of these groups reacts with DTNB . By contrast, all of these groups (11/subunit) of the native chicken liver pyruvate carboxylase are accessible to DTNB . The thermophile enzyme is also more resistant to other sulfhydryl reagents and to denaturation under certain conditions than the avian enzyme.

Biochim Biophys Acta, 1979 Aug 15, 569(2), 239 - 48
Ribulose 1,5-bisphosphate carboxylase from the thermophilic, acidophilic alga, Cyanidium caldarium (Geitler) . Purification, characterisation and thermostability of the enzyme; Ford TW; An an initial stage in the study of proteins from thermophilic algae, the enzyme ribulose 1,5-bisphosphate carboxylase 2-phospho-D-glycerate carboxylyase (dimerizing, EC 4.1.1.39) was purified 11-fold from the thermophilic alga Cyandium caldarium, with a 24% recovery . This purified enzyme appeared homogeneous on polyacrylamide gels and could be dissociated into two subunit types of molecular weights 55,000 and 14,900 . The optimal assay temperature was 42.5 degrees C, whilst enzyme purified from Chlorella spp . showed maximum activity at 35 degrees C . The thermostability of Cyanidium ribulose 1,5-bisphosphate carboxylase was considerably greater than that of the Chlorella enzyme, and the presence of Mg2+ and HCO-3 further enhanced this heat stability . A break in the Arrhenius plot occured at 20 degrees C for Chlorella ribulose 1,5-bisphosphate carboxylase and 36 degrees C for the enzyme from Cyanidium . It is suggested that the thermostability of Cyanidium ribulose 1,5-bisphosphate carboxylase is a result of an inherent stability of the enzyme molecule which permits efficient CO2 fixation at high temperatures but results in low activity in the mesophilic temperature range.

Genetics, 1979 Aug, 92(4), 1079 - 92
An enrichment for temperature-sensitive mutants in Tetrahymena thermophila; Martindale DW et al.; The parameters for the killing of Tetrahymena by 5-bromodeoxyuridine(BUdR) and near-ultraviolet light have been determined . Significant preferential killing by UV of cells that have incorporated BUdR was obtained when the cells were irradiated in a nonnutrient buffer . UV alone was found to be toxic to cells irradiated in growth medium . Mutants defective in division at a restrictive temperature were isolated from mutagenized cultures that had been treated with BUdR and UV and from mutagenized cultures that had no such treatment . Results indicate that the number of temperature sensitive (ts) growth mutants can be increase five to six times using the BUdR/UV treatment . Data are presented that indicate differences in the frequency of occurrence of various types of ts mutants, with and without enrichment . A mutant that immediately stopped macromolecular synthesis and cell division upon being placed at the restrictive temperature was more resistant to BUdR/UV treatment than wild type by 1000-fold . Using the above techniques, BUdR-resistant mutants altered in the phosphorylation of thymidine have been isolated.

Int J Pept Protein Res, 1979 Aug, 14(2), 99 - 106
Comparative temperature-stability properties of malate dehydrogenases from some thermophilic fungi; Wali AS et al.; Temperature-activity and temperature-stability relationships of malate dehydrogenases from eight thermophilic fungi were determined . Temperature optima for maximum activity of the enzymes varied between 50 degrees and 60 degrees and the Arrhenius plots were linear between 5 degrees and 50 degrees . The energies of activation ranged from 2.1 Kcal/mol for the enzyme from Sporotrichum thermophile to 9.1 Kcal/mol for that from Penicillium duponti . Heat inactivation kinetics at 50 degrees revealed heat lability of the enzyme from most of the thermophilic fungi . The t1/2's (min) were less than 10 for the enzymes from P . duponti, S . thermophile and Thermoascus aurantiacus; less than 30 for those from Chaetomium thermophile var . coprophile, H . lanuginosa and C . thermophile var . dissitum; and greater than 30 for those from Mucor pusillus and H . insolens . Salts of Na+, K+ and NH4+, and citrate protected the enzymes from H . lanuginosa, C . thermophile var . dissitum and M . pusillus against heat inactivation.

J Protozool, 1979 Aug, 26(3), 519 - 24
Acid hydrolases and their release in food vacuole-less mutants of Tetrahymena thermophila; Silberstein GB; Mutants (NP1 and PSJ5) of Tetrahymena thermophila strains B and D 1968 exist that are unable to construct a functional oral apparatus and form food vacuoles at 37 C but which do so normally at 30 C . Food vacuole-less cells starved in dilute salt solution released similar amounts of acid phosphatase, beta-N-acetyl-glucosaminidase and alpha-glucosidase activity into the medium as wildtype cells during an 8-h period . Actively growing, food vacuole-less cells had approximately 50% less total protein, acid phosphatase, beta-N-acetyl-glucosaminidase, and alpha-glucosidase per cell than wildtype cells after 72-h growth . During this time food vacuole-less cells released significant amounts of the 3 acid hydrolases into the growth medium . For each hydrolase, the total activity released from growing, food vacuole-less cells was less, on a per cell basis, tahn the amount released from food vacuole formers . The proportion of the total activity secreted by the mutant and the wildtype cells was the same for acid phosphatase and beta-N-acetyl-glucosaminidase and somewhat lower for alpha-glucosidase . It is concluded that the release of a significant amount of acid hydrolase activity from Tetrahymena is independent of food vacuole formation and may be analogous to the secretory activity of other nonphagocytic eukaryotic cells.

J Bioenerg Biomembr, 1979 Aug, 11(3-4), 39 - 78
Structure and function of H+-ATPase; Kagawa Y et al.; (1) Extensive studies on proton-translocating ATPase (H+-ATPase) revealed that H+-ATPase is an energy transforming device universally distributed in membranes of almost all kinds of cells . (2) Crystallization of the catalytic portion (F1) of H+-ATPase showed that F1 is a hexagonal molecule with a central hole . The diameter of F1 is about 90 A and its molecular weight is about 380,000 . (3) Use of thermophilic F1 permits the complete reconstitution of F1 from its five subunits (alpha, beta, gamma, delta, epsilon) and demonstration of the gate function of the gamma delta epsilon-complex, the catalytic function of beta (supported by alpha and gamma), and the H+-translocating functions of all five subunits . (4) Studies using purified thermostable F0 showed that F0 is an H+-channel portion of H+-ATPase . The direct measurement of H+-flux through F0, sequencing of DCCD-binding protein, and isolation of F1-binding protein are described . (5) The subunit stoichiometry of F1 may be alpha 3 beta 3 gamma delta epsilon . (6) Reconstitution of stable H+-ATPase-liposomes revealed that ATP is directly synthesized by the flow of H+ driven by an electrochemical potential gradient and that H+ is translocated by ATP hydrolysis . This rules out functions for all the hypothetical components that do not belong to H+-ATPase in H+-driven ATP synthesis . The roles of conformation change and other phenomena in ATP synthesis are also discussed.

Biochim Biophys Acta, 1979 Jul 11, 569(1), 6 - 12
Activation of Thermus phosphofructokinase by monovalent cations; Stellwagen E et al.; The presence of the monovalent cations Tl+, NH+4, K+, Rb+ or Cs+, in decreasing order of potency, produce a marked equivalent increase in the specific enzyme activity of phosphofructokinase (ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) purified from extreme thermophile, Thermus X-1 . By contrast, the monovalent cations Li+, Na+ or CH3NH+3 produce no detectable catalyitic activation at concentrations up to 100 mM . The relative potency of these cations suggests that each polypeptide chain in the tetrameric enzyme possesses a cationbinding site having tetragonal symmetry and that the protein ligands are principally hydroxyl or carboxylate oxygens . Only the enzyme-cation complex and not the enzyme by itself exhibits cooperativity with respect to the dependence of catalytic rate on the concentration of the substrate, fructose 6-phosphate . In the presence of subsaturating but not saturating concentrations of substrate, the catalytic activation produced by monovalent cations is also cooperative . Exclusion chromatographic measurements indicate that the enzyme remains tetrameric at catalytic concentrations in the presence or absence of an activating monovalent cation.

Appl Environ Microbiol, 1979 Jul, 38(1), 166 - 8
Distribution of Aeromonas hydrophila in natural and man-made thermal effluents; Hazen TC et al.; Densities of Aeromonas hydrophila showed distinct thermal optima (25 to 35 degrees C) and thermal maxima (45 degrees C) when measured along thermal gradients created by geothermal and nuclear reactor effluents . Survival of A . hydrophila never exceeded 48 h at temperatures of greater than 45 degrees C . Thermophilic strains could not be isolated at any site.

J Biochem (Tokyo), 1979 Jul, 86(1), 121 - 30
Effects of divalent cations on thermophilic inorganic pyrophosphatase; Hachimori A et al.; Divalent cations were shown to affect the structure and thermostability of thermophilic inorganic pyrophosphatase {pyrophosphate phosphohydrolase EC 3.6.1.1} purified from Bacillus stearothermophilus and thermophilic bacterium PS-3 . The properties of the enzymes from the two sources were found to be very similar . The enzymes were very unstable to heart in the absence of divalent cations, being inactivated gradually even at 40 degrees C . However, they became stable to heat denaturation in the presence of Mg2+, between pH 7.8 and 9.0 . Similar induced thermostability was detected when Mn2+, Co2+, Ca2+, Cd2+, and ZN2+ were added, though the latter three cations were not essential for enzyme activity . On adding divalent cations, the optical properties such as absorption spectra, fluorescence spectra, and circular dichroism (CD) were changed . Gel filtration and disc electrophoresis revealed that the molecular weight of both enzymes was 5.4 x 10(4) in Tris-SO4 buffer and 11 x 10(4) in Tris-HCL buffer, suggesting monomer-dimer transformation . In the presence of divalent cations in Tris-SO4 fuffer, the enzymes dimerized; this was confirmed by sedimentation velocity measurements . The enzymes in Tris-HCL buffer did not show thermostability unless divalent cations were added . The results in the present study indicate that binding of divalent cations to each enzyme caused some conformational change in the vicinity of aromatic amino acid residues leading to dimerization of the enzyme molecule so that it became thermostable . It was also suggested that histidyl residues play an important role in the thermostability induced by divalent cations on the basis of the pH dependencies of thermostability and CD spectra.

Hoppe Seylers Z Physiol Chem, 1979 Jul, 360(7), 795 - 807
Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria . I) Isolation and characterization of lactate dehydrogenases from thermophilic and mesophilic bacilli; Schar HP et al.; Lactate dehydrogenases from thermophilic bacilli (Bacillus stearothermophilus, Bacillus caldotenax) and from mesophilic bacilli (Bacillus X1, Bacillus subtilis) have been isolated by a two-step purification procedure . Only one type (LDH-P4) composed of four identical subunits (Mr 34 000 or 36 000) was found in each bacillus . The tetrameric enzymes were characterized with respect to thermostability, pH and temperature dependence of the pyruvate reduction and the L-lactate oxidation, substrate specificity, saturation kinetics (Km values of pyruvate, lactate, NAD, NADH), pyruvate and oxamate inhibition, and activation by fructose bisphosphate . The thermophilic and mesophilic enzymes differ characteristically in these parameters . Preliminary structural data (amino acid composition, comparative N-terminal sequence analysis) show the expected close phylogenetic relationship (high degree of sequence homology), but also typical differences between thermophilic and mesophilic dehydrogenases, a suitable basis for further comparative studies.

Rev Bras Pesqui Med Biol, 1979 Jun, 12(2-3), 189 - 91
Thermophilic Campylobacter-associated diarrhoea in Rio de Janeiro; Ricciardi ID et al.; Thermophilic Campylobacter is reported as the agent associated with chronic and acute diarrhoea in Rio de Janeiro . Nine strains were isolated from 186 children with gastroenteritis . To our knowledge this is the first report of human Campylobacteriosis in Latin-America.

J Gen Microbiol, 1979 Jun, 112(2), 357 - 64
The control of the synthesis of isocitrate lyase in a thermophilic bacillus; Griffiths MW; From a strain of Bacillus stearothermophilus, devoid of active pyruvate carboxylase, a mutant (NG-15) was selected that grew on acetate in the presence of glucose . This mutant differed from its parent organism in possessing high activities of isocitrate lyase when grown on all carbon sources tested except nutrient broth, in possessing unusually low activities of NADP+-dependent isocitrate dehydrogenase and in containing increased amounts of isocitrate . Revertants of mutant NG-15 which regained the ability to synthesize active pyruvate carboxylase also synthesized isocitrate lyase and isocitrate dehydrogenase to the same extent as the wild-type strain . These results suggest that the regulatory mechanism for the synthesis of isocitrate lyase in the thermophile may be different from that in mesophilic bacilli.

J Biochem (Tokyo), 1979 Jun, 85(6), 1509 - 17
Purification and properties of phosphoglycerate kinase from Thermus thermophilus strain HB8; Nojima H et al.; (1) A glycolytic enzyme, phosphoglycerate kinase {EC 2.7.2.3}, was purified from cells of an extreme thermophile, Thermus thermophilus strain HB8 . The enzyme was resistant to heat, and no loss of activity was observed after incubation for 10--20 min at 79 degrees C . (2) Catalytic properties such as pH optimum (pH 6--8.5), kinetic parameters (Km=0.28 mM for ATP, 1.79 mM for glycerate 3-phosphate), substrate specificity and inhibitors of the enzyme were investigated and compared with those of phosphoglycerate kinase from other sources . (3) The enzyme protein consists of a single polypeptide chain of molecular weight 44,600 . The isoelectric point is 5.0 The amino acid composition of the enzyme was studied . The contents of ordered secondary structures were estimated to be 29% alpha-helix and 11% pleated sheet from the circular dichroic spectrum of the enzyme protein . (4) The fluorescence spectrum of the enzyme protein showed an emission maximum at 320 nm when excited at 280 nm . The quantum yield was 0.19 . Tryptophyl fluorescence was not quenched, in contrast to the fluorescence reported for yeast phosphoglycerate kinase.

J Bacteriol, 1979 Jun, 138(3), 903 - 8
Use of 8-hydroxyquinoline to enrich for temperature-sensitive mutants of Tetrahymena; Martindale DW et al.; The effects of the chelating agent 8-hydroxyquinoline (Hq) on Tetrahymena thermophila were examined . Cell division was completely inhibited by 5 micrograms of Hq per ml . At this concentration deoxyribonucleic acid, ribonucleic acid, and protein syntheses were also completely and nonselectively inhibited . The inhibition was reversible after 6 h of Hq treatment . At concentrations above 20 micrograms/ml a 10,000-fold decrease in survival as seen after 2 h in the drug . The sensitivity of Tetrahymena to Hq was found to be dependent upon cell concentration, wild-type strain, medium, and length of time the culture is at 38 degrees C before Hq is added . Mutants of Tetrahymena that are unable to divide at the restrictive temperature, but which continue macromolecular synthesis, were found to be resistant to Hq treatment . Conditions were obtained in which more than a 1,000-fold difference in survival was seen between this class of mutant and the wild type . The effect of Hq on three other classes of temperature-sensitive mutants was examined, and the results are discussed.

Sabouraudia, 1979 Jun, 17(2), 163 - 9
Antigenic relationships among thermophilic actinomycetes; Kurup VP et al.; Antigens from Micropolyspora faeni, Saccharomonospora viridis, Thermoactinomyces candidus, T . sacchari and T . vulgaris were prepared by growing them on dialysate of trypticase soy broth . Sera from rabbits immunized with these antigens were used to study cross-reactivity between thermophilic actinomycetes by antigen-antibody crossed immunoelectrophoresis and by agar gel double diffusion . Mi . faeni and S . viridis showed some degree of cross-reaction, but both failed to show any cross reactivity with Thermoactinomyces species . Antigens from Thermoactinomyces cross-reacted with members within the genus, but no reactivity against S . viridis or M . faeni antisera was detected . Hence, the presence of antibodies to several thermophilic actinomycetes in the sera of patients may be attributed to the exposure of the individual to different thermophilic actinomycetes rather than to the antigenic cross-reactivity between the organisms.

Ann Microbiol (Paris), 1979 May-Jun, 130 A(4), 487 - 98
{Amoebae in the water supply: an epidemiological study (author's transl)}; Dive D et al.; Drinking water samples (836) of various origins have been examined for amoebae . For collector water, half of the samples are contamined without influence of the origin . So, occurrence of amoeba in underground waters is discussed . Filtration and disinfection are generally inefficient . Bottles of drinking water are very often polluted during bottling . International mineral waters of french origin are not so damaged during conditioning operation . Bacteria and amoebae populations are quite independant and can't be indicators one for another . Thermophilic amoebae strains are very rare and any pathogenic amoeba has been found during the study.

J Protozool, 1979 May, 26(2), 241 - 5
Selection of nonmotile Tetrahymena with Ficoll underlayers; McGwin NF et al.; The mechanisms regulating the development of cilia in Tetrahymena are poorly understood but might be revealed through the study of ciliogenesis mutants . Failure to regenerate cilia after dibucaine deciliation results in continued absence of motility . Therefore, to isolate ciliogenesis mutants efficiently, methods for separating motile and nonmotile cells are essential . We examined the efficacy of Ficoll underlayers for these separations . Ciliates of T . thng type IV) were mixed with Ficoll and added as underlayers to separatory funnels containing growth medium . At 27 C most of the cells remained motile and were found in the top layer; at 37 C, there was a time-dependent increase in the number of nonmotile cells and the number of cells in the Ficoll layer . After 150 min at 37 C, most of the cells became nonmotile and were found in the Ficoll layer . Other studies indicated that at 37 C, the cells remained alive and capable of regenerating cilia when deciliated . Thus, it is clear that the Ficoll underlayer effectively separates the majority of nonmotile cells from the majority of motile cells . Evidently, however, at 37 C wild-type T . thermophila exhibit temperature-sensitive phenotypic variability with regard to motility which should be minimized when selecting for mutations affecting motility and ciliogenesis.

J Biochem (Tokyo), 1979 May, 85(5), 1205 - 11
Bacteriophage phiNS11: a lipid-containing phage of acidophilic thermophilic bacteria . IV . Sedimentation coefficient, diffusion coefficient, partial specific volume, and particle weight of the phage; Sakaki Y et al.; The particle weight (molecular weight) of phiNS11 was determined from the sedimentation coefficient, diffusion coefficient, and partial specific volume of the phage . The sedimentation coefficient of the phage (S(0)20, W) is 416 +/- 2.7S . The diffusion coefficient D(0)20, W), which was determined by quasielastic light scattering measurement, is (0.57 +/- 0.03) x 10(-7) cm2/s . The partial specific volume was determined by the mechanical oscillation technique to be 0.747 +/- 0.007 cm3/g . Based on these values, the particle weight of the phage was calculated to be (70.3 +/- 4.3) x 10(6) daltons, which agrees well with the particle weight (69--72 x 10(6) daltons) estimated from the molecular weight of phage DNA and the content of DNA . The Stokes radius of the phage particle was calculated to be 37.7 +/- 2 nm and hydration of the phage was estimated to be 1.18 cm3/g of dry phage . From the particle weight and the chemical composition of the phage, we estimated that one phage particle contains one double-stranded DNA molecule, 16,000 residues of fatty acid, 72 protein I molecules, 920 protein II, 42 protein III, 48 protein IV, 290 protein V molecules, and 3,700 molecules of polyamines.

Biochem J, 1979 May 1, 179(2), 407 - 12
Adenosine triphosphate consumption by bacterial arginyl-transfer ribonucleic acid synthetases; Godeau JM et al.; ATP consumption by arginyl-tRNA synthetases from Escherichia coli and Bacillus stearothermophilus has been investigated by the firefly luciferin--luciferase assay . Arginyl-tRNA synthetase from E . coli utilizes ATP only for aminocylation of tRNA with a 1:1 stoicheiometry . In contrast, we have shown an adenosine triphosphatase activity of arginyl-tRNA synthetase from B . stearothermophilus in the absence of tRNAArg . Dowex chromatography revealed the formation of ADP by the thermophile enzyme; under aminoacylation conditions, AMP was also formed in amounts stoicheiometric with arginyl-tRNA formation.

Nucleic Acids Res, 1979 Apr, 6(4), 1571 - 81
Role of ribothymidine in the thermal stability of transfer RNA as monitored by proton magnetic resonance; Davanloo P et al.; In order to elucidate the functional role of the modified uridines at position 54 of tRNA, the 270 MHz high-field proton NMR spectra of methionine tRNAs from E . coli, from a mutant thereof, and from T . thermophilus, containing ribothymidine, uridine and 2-thioribothymidine, respectively, have been measured as a function of temperature . A comparison of the NMR melting profiles of the minor nucleosides from these tRNAs shows that the melting temperature of uridine containing tRNA is 6 degrees C lower than that of the wild type tRNA whereas that of the 2-thioribothymidine tRNA is 7 degrees C higher than that of the wild type tRNA . These results, therefore, demonstrate that these modifications serve for stabilization of the tertiary structure of tRNA.

J Cell Sci, 1979 Apr, 36, 343 - 53
Dual capacity for nutrient uptake in Tetrahymena . V . Utilization of amino acids and proteins; Orias E et al.; We investigated the relative contributions of phagocytosis and plasma membrane transport to the uptake of amino acids and a protein (egg albumin) in amounts which allow Tetrahymena thermophila to grow and multiply . We used a mutant capable of indefinite growth without food vacuole formation (phagocytosis) and its wild type (phagocytosis-competent) isogenic parental strain . Our results suggest that phagocytosis is not required for free amino acid uptake, most or all of which can be attributed to carrier-mediated transport systems, apparently located on the plasma membrane . In contrast, phagocytosis is required for utilization of the protein . Proteins can supply required amino acids in amounts sufficient for growth only when food vacuoles are formed . We conclude that Tetrahymena thermophila either possesses no endocytic mechanisms at the cell surface other than food vacuole formation or, if it does, these putative mechanisms are not capable of nutritionally meaningful rates of protein uptake.

Prax Klin Pneumol, 1979 Apr, 33 Suppl 1, 640 - 3
{Prevention and clinical aspects of pneumoconiosis induced by organic dusts (author's transl)}; Antweiler H; Only those pneumoconioses induced by organic dusts are discussed which are met with in Germany . They comprise: byssinosis which is caused by toxic agents in cotton and flax, farmer's lung, sprayer's fever, exogenous allergic alveolitis due to thermophil actinomyces in the dust of mouldy agricultural meterials or the air of air-conditioning systems, bird fancier's lung which is caused by antigens in animal waste products . Byssinosis and farmer's lung have now been acknowledged as occupational diseases.

Mikrobiologiia, 1979 Mar-Apr, 48(2), 222 - 5
{tRNA-methylase study of the extreme thermophile, Thermus flavus}; Shershneva LP et al.; tRNA methylases were studied in the extreme thermophilic culture of Thermus flavus, strain 71 . Like E . coli, the culture contained only those tRNA methylases which catalysed the formation of m1A and m7G . Mg2+, Ca2+ and Na+ ions activated tRNA methylases of Thermus flavus in the series Mg greater than Ca greater than Na while Mn2+ ions inhibited the enzyme . The activity of tRNA methylases was higher in T . flavus than in E . coli, and required less protein and time for exhaustive methylation of tRNA preparations . The overall activity of methylases in T . flavus at 70 degrees C was 5-6 times higher than at 40 degrees C; the elevation of temperature had different effect on various methylases: the activity of m1A methylase increased 13-fold whereas that of m7G methylase only twofold.

Biotechnol Bioeng, 1979 Mar, 21(3), 345 - 55
Association of beta-glucosidase with intact cells of Thermoactinomyces; Hagerdal B et al.; The location of the B-glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied . Little beta-glucosidase activity was found in the culture filtrate, while the culture solids contained the major part of the activity of the whole culture broth . The activity does not appear to be adsorbed to the culture solids; rather there is evidence that it is an intracellular soluble enzyme(s) . The pH and temperature optima for a crude beta-glucosidase preparation were determined to be pH 6.5 and 50--55 degrees C . Enzyme activity studies indicate that the same enzyme(s) accounts for the beta-glucosidase and the cellobiase activities . The validity of using the filter paper activity of culture filtrates from Thermoactinomyces to predict the total saccharification of cellulosic materials to glucose is discussed.

Vet Med (Praha), 1979 Mar, 24(3), 185 - 92
{Microbiology of various liver products}; Lukasova J et al.; The proportions of mesophilic, thermophilic and psychrophilic microorganisms were investigated in the total number of microbes and aerobic sporogenic microorganisms in delicatessen liver sausages and liver cheeses . At the same time the content of ammonia and the pH of the products were determined . The largest part of the microflora comprised mesophilic microorganisms, 10 to 25% of which belonged to the sporogenic microflora . The incidence of staphylococci, enterococci and coliform microorganisms was also studied . The total number of microbes contributed to the ammonia content in the products, however, no intercorrelation was statistically proved . In the case of pH no relationship between its value and the number or kind of investigated microorganisms was found . All the indicators were followed in the raw material, in the finished products after heat processing and in products stored at refridgerator and room temperatures for one week.

J Am Vet Med Assoc, 1979 Feb 1, 174(3), 277 - 81
The role of allergy in chronic pulmonary disease of horses; Halliwell RE et al.; Twenty-five horses with chronic pulmonary disease were skin tested with allergenic extracts of 24 molds, 4 thermophilic actinomyces, barn dust, hay dust, soya-bean mill dust, and grain mill dust . The results were compared with those obtained on 25 normal horses . Between the 2 groups of horses, there was a highly significant difference in positive skin test results at 30 minutes and 4 hours.

Biochem J, 1979 Feb 1, 177(2), 441 - 8
Simple efficient methods for the isolation of malate dehydrogenase from thermophilic and mesophilic bacteria; Wright IP et al.; Malate dehydrogenase from a number of bacteria drawn from several genera and representing the mesophilic, moderately thermophilic and extremely thermophilic classes was isolated by procedures which involve only a small number of steps (in most cases only two), of which the key one is affinity chromatography on 5'-AMP--Sepharose and/or on NAD+--hexane--agarose . Electrophoretic analysis of the native enzymes in polyacrylamide gel and of the denaturated enzymes in sodium dodecyl sulphate/polyacrylamide gel revealed no significant protein impurity in the purified preparations . The yields ranged from about 40% to over 80% . The malate dehydrogenases from the extreme thermophiles and from some of the moderate thermophiles are appreciably less efficient catalytically than their mesophilic homologues.

J Biochem (Tokyo), 1979 Feb, 85(2), 503 - 9
Carbodiimide-binding protein of H+-translocating ATPase and inhibition of H+ conduction by dicyclohexylcarbodiimide; Sone N et al.; H+-Translocating ATPase, which catalyzes ATP synthesis in biomembranes, is composed of a head piece (F1) and a membrane moiety (F0) . Using highly-purified F0 from a thermophilic bacterium PS3 (TF0), the following results were obtained . 1 . Inhibition by N,N'-dicyclohexylcarbodiimide (DCCD) of H+ conduction through TF0 followed pseudo-first-order kinetics . The second-order rate constant for inhibitor-enzyme interaction was 5 times 10(3) M(-1)-min(-1) . 2 . H+ conductivity blocked by DCCD was proportional to the amount of DCCD incorporated in the band 8 protein of TF0 . When only one-third of the band 8 protein was labeled with DCCD, TF0 hardly transported any H+ . 3 . By extracting TF0 with chloroform-methanol, the band 8 protein was obtained as a proteolipid . Polyacrylamide gel electrophoresis with dodecyl sulfate and urea showed that the molecular weight was about 6,000 . 4 . The amino acid composition of band 8 protein indicated that this protein contained an extremely high percentage of hydrophobic amino acids (0.29 in polarity) and was devoid of histidine, tryptophan, cysteine, and lysine . Its minimum molecular weight was 6,500 . 5 . The role of band 8 protein (DCCD-binding protein) in H+ conduction through TF0 is discussed on the basis of these results.

Biochim Biophys Acta, 1979 Jan 12, 566(1), 62 - 6
Hydrolysis of low molecular weight isomaltosaccharides by a p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from a thermophile, Bacillus thermoglucosidius KP 1006; Suzuki Y et al.; A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase of a thermophile, Bacillus thermoglucosidius KP 1006, was purified to an electrophoretically-homogeneous state . Its molecular weight was estimated as 60 000 by gel electrophoresis . The molecular activity (ko) and the Km value at 60 degrees C and pH 6.8 for p-nitrophenyl-alpha-D-glucopyranoside were 233 s-1 and 0.24 mM, respectively . The enzyme cleft the non-reducing terminal alpha-1,6-glucosidic bonds of isomaltose, panose, isomaltotriose, isomaltotetraose, and isomaltopentaose . The ko values were 72.4, 194, 208, 233 and 167 s-1, and the Km values were 3.3, 9.5, 11, 13 and 21 mM, respectively . Each isomaltosaccharide was hydrolyzed to glucose by the cleavage of single glucose units from its nonreducing end . The present study suggests that the enzyme is an oligo-1,6-glucosidase (dextrin 6-alpha-glucanohydrolase, EC 3.2.1.10) and an exo-glucosidase.

Chromosoma, 1979, 75(3), 293 - 308
An analysis of spindle ultrastructure during prometaphase and metaphase of micronuclear division in Tetrahymena; LaFountain JR Jr et al.; Mitotic micronuclei were isolated from Tetrahymena thermophila in a medium containing hexylene glycol and their ultrastructure was analyzed using thin section techniques . The two stages selected for analysis were early prometaphase and metaphase . A comparison of data from these two stages revealed several differences in nuclear morphology . Metaphase nuclei were longer, they contained more microtubules, and the distribution of microtubules at metaphase was different from that at early prometaphase . Increases in microtubules, which are a unique class of microtubules that can be distinguished from other classes on the basis of their close association to the nuclear membrane . Growth of peripheral sheath microtubules is thought to be significant because it could be the mechanical basis of nuclear elongation . Crossbridges were observed throughout the spindle between all classes of microtubules, but the exact function of these elements remains to be determined.

Folia Microbiol (Praha), 1979, 24(5), 396 - 402
Thermomonospora sp . T-SA-125 and its production of a growth promoting antibiotic; Dewedar A et al.; Thermomonospora sp . T-SA-125 is a true thermophilic actinomycete isolated from a soil sample collected from the Saudi Arabian desert . It is characterized by the formation of single spores at the tips of dichotomously branched aerial mycelium and differs from Thermomonospora curvata and T . viridis in certain aspects . It produces a basic water-soluble antibiotic which is active against Gram-positive bacteria, moderately active against Gram-negative bacteria and inactive against fungi . At high concentrations, this antibiotic, stimulated the growth of both Hordeum coleoptile and lettuce hypocotyl.

J Embryol Exp Morphol, 1979 Jan, 49, 203 - 27
A mutant of Tetrahymena thermophila with a partial mirror-image duplication of cell surface pattern . II . Nature of genic control; Frankel J et al.; The CU-127 clone of Tetrahymena thermophila, which manifests an unusually high number of ciliary rows plus a second set of abnormal oral structures and of contractile vacuole pores with partial mirror-image reversal of asymmetry (Jerka-Dziadosz & Frankel, 1979), has been subjected to breeding analysis . The progeny ratios obtained in various crosses indicate that the abnormalities of cell-surface asymmetry are brought to expression as a result of the action of a recessive allele at a single gene locus, here named janus . When previously normal cells were made homozygous for the jan allele, the cortical pattern characteristic of the CU-127 clone came rapidly to expression, often without associated change in number of ciliary meridians . Conversely, when cells previously expressing jan re-acquired the wild-type (jan+) allele, they returned to the normal pattern of a single normal oral structure and a single normally located set of contractile vacuole pores while still retaining the high ciliary meridian number characteristic of the original CU-127 clone . The capacity for manifestation of the unique asymmetry pattern depends solely on homozygous expression of the janus allele, whereas the stable number of ciliary meridians in janus clones and the degree of expression of secondary OAs is modulated by other factors, probably at least in part genic . These results, taken together with those of the preceding paper, indicate that the janus allele promotes the propagation and/or expression of a condition of reversed asymmetry in a precisely located cell region, and further indicates that the propagation and expression of this condition are largely independent of the number and asymmetry of ciliary meridians.

J Embryol Exp Morphol, 1979 Jan, 49, 167 - 202
A mutant of Tetrahymena thermophila with a partial mirror-image duplication of cell surface pattern . I . Analysis of the phenotype; Jerka-Dziadosz M et al.; Cells of a mutant clone, CU-127, of Tetrahymena thermophila (formerly T . pyriformis, syngen 1) manifest three anatomical abnormalities . First, the stable number of ciliary meridians is 21-25, above the usual number (17-21) in this species . Second, up to 30% of the cells have two oral apparatuses (OAs), one normal and the other abnormal . Third, more than one-half of the cells possess two distinct sets of contractile vacuole pores (CVPs) . In some living cells two contractile vacuoles are seen . These abnormalities have persisted unchanged during more than 500 generations of vegetative propagation, and are similarly expressed in subclones . The normal and abnormal OAs are topographically segregated, with normal OAs developing along the "primary oral axis" and abnormal OAs developing along a "secondary oral axis" that is situated 170 degrees of the cell circumference to the cell's right of the primary oral axis . CVPs always appear within this 170 degree arc and never within the complementary 190 degrees arc to the left of the primary oral axis . A unique feature of the CU-127 clone is the commonly expressed mirror image reversal of the structural pattern of OAs that develop along the secondary oral axis . The primordia of such OAs initially appear (as usual) to the cell's left of a ciliary meridian, but as membranelles develop the frequently come to be oriented in a mirror image of the normal pattern, and an undulating membrane sometimes develops on the wrong (left) side of the oral primordium . When two sets of CVPs are formed, their average positions are roughly equidistant with respect to the two oral axes, with the two sets located 50-60 degrees to the right and left respectively of the primary and secondary oral axis . Such cells are thus bilaterally symmetrical about a plane defined by the central longitudinal axis and the halfway point between the two CVP sets (see Fig . 25) . This plane bisects the cell into a normal and a "reversed" half-cell . However, only oral asymmetry and large-scale CVP positioning are subject to such reversal; all ciliary meridians remain of normal asymmetry and all CVPs are situated on the left side of CVP meridians . The fact that major aspects of large-scale cellular organization can be reversed while the "fine-positioning" associated with the ciliary meridians remains normal indicates that the two aspects of cell organization are distinct.

Mikrobiologiia, 1979 Jan-Feb, 48(1), 133 - 6
{Fixation of molecular nitrogen by sulfate-reducing bacteria from petroleum strata}; Nazina TN et al.; The activity of nitrogen fixation by the museum strains of sulfate reducing bacteria isolated from oil deposits was studied using the acetylene technique . The mesophilic sulpfate reducing bacteria belonging to the species Desulfovibrio africanus 2372 and D . baculatus X were found to have a high nitrogenase activity . D desulfuricans subsp . aestuarii 2198 reduced acetylene at a low rate . The thermophilic sulfate reducing cultures of D . thermophilus 7, Desulfotomaculum nigrificans 781 and Dm . nigrificans subsp . salinus 435 produced only small quantities of ethylene . Apparently, nitrogen fixation by sulfate reducing bacteria can be considerable in oil strata whose temperature does not exceed 35--40 degrees C.

Clin Allergy, 1979 Jan, 9(1), 43 - 52
Microbiological and serological studies of farmers' lung in Finland; Terho EO et al.; Sera from few Finnish patients with clinical farmers' lung react in precipitin tests with extracts of the thermophilic actinomycetes that commonly cause the disease elsewhere . Hays associated with the disease in Finland showed less evidence of spontaneous heating and contained fewer actinomycete spores than British hays . Only Thermoactinomyces vulgaris was sometimes abundant . Some species of mesophilic fungi were more abundant than in Britain and one, Aspergillus umbrosus, reacted with most sera from farmers' lung patients in precipitin tests . A panel of antigens, including thermophilic actinomycetes, A . umbrosus and other species of the Aspergillus glaucus group, is recommended for screening farmers' lung sera.

Antonie Van Leeuwenhoek, 1979, 45(2), 303 - 12
Effects of fungicides on Aspergillus fumigatus; Kuthubutheen AJ et al.; Aspergillus fumigatus was the most frequently isolated thermophilous fungus from green leaf surfaces . The application of fungicides significantly reduced the frequency of its occurrence there . A . fumigatus was relatively tolerant to fungicides . On Captan-, Thiram-, and Verdasan-treated leaves, A . fumigatus constituted 66%--80% of the total number of isolates obtained at 45 degrees C from each treatment while Dicloran did not depress the percentages . At 45 degrees C, A . fumigatus was found to be strongly cellulolytic with a slow rate of radial extension on YpSs agar and rapid rate of mycelial growth in Czapek Dox liquid medium . Increasing concentrations of all four fungicides reduced or prevented growth, sporulation, starch depletion and cellulose clearing of A . fumigatus . The fungus could tolerate higher concentrations of HgCl2 than of Verdasan . 2.5 microgram/ml of the four fungicides altered the rates of mycelial growth but not the maximum amount of mycelial dry weight attained.

Respiration, 1979, 37(1), 52 - 60
Farmer's lung disease in Vermont; Gump DW et al.; Several Vermont population groups were surveyed for the occurrence of antibodies to thermophilic actinomycetes . Antibodies to M . faeni and T . vulgaris were measured by the precipitin method in all subjects and, in 124 subjects, M . faeni antibodies were also measured by the indirect fluorescent antibody (IFA) technique . There was relatively good correlation between the two techniques (r 0.48, p less than 0.01) . Hospital employees, blood donors and patients with chronic bronchitis were generally negative for precipitins to thermophilic actinomycetes . Of the 258 Vermont dairy farmers surveyed, 14 (5.4%) had precipitins to M . faeni, 3 (1.2%) had precipitins to T . vulgaris but only 1 farmer with antibodies to M . faeni had symptoms of possible farmer's lung disease (FLD) . On the other hand, 10 (4.1%) precipitin-negative farmers had symptoms possibly consistent with FLD . The IFA test did not correlate any better with symptoms . 7 (5.6%) patients with pulmonary fibrosis had precipitins to M . faeni 5 of these were diagnosed as having FLD . 18 (14.4%) had precipitins to T . vulgaris and only 3 of these patients were felt to have hypersensitivity pneumonitis . Patients with pulmonary fibrosis and FLD hat IFA titers of greater than or equal to 1/128, but so did asymptomatic farmers.

Nucleic Acids Symp Ser, 1979, (6), s135 - 8
An endonucleolytic activity of nuclease TT1 specific for superhelical DNA; Takahashi M et al.; Highly purified nuclease TT1 from T . thermophilus HB8 acts on a linear single- and double-stranded DNA as an exonuclease and produces 5'-mononucleotides either from the 5'- or 3'-terminus . It was found that the enzyme also possesses an endonuclease activity specific for superhelical (form I) and single-stranded circular DNA . Form I of various kinds of DNA (phi X174, PM2, Co1E1 and RF 1010 etc.) is nicked to yield first relaxed circles (form II) and then nicked at the opposite site to yield unit length linear DNA (form III), which is subsequently hydrolyzed from the 5'- or 3'-terminus . A single cleavage of the form I of phi X174 DNA seemed to occur at a limited number of unique sites . Both endonuclease and the known exonuclease activities co-migrate on polyacrylmide gels, show the same pH and temperature optima, are stimulated by Mg2+ and are inactivated by EDTA similarly.

Mikrobiologiia, 1979 Jan-Feb, 48(1), 80 - 9
{Localization of hydrogenase in the cells of the thermophilic hydrogen bacterium, Pseudomonas thermophila}; Emnova EE et al.; Spheroplasts that were osmotically stable in 0.2M Tris-HCl--0.02M EDTA were prepared from the autotrophically grown cells of Pseudomonas thermophila K-2 . The spheroplasts possessed 90--95% of the hydrogenase activity of the whole cells . The half-life time of hydrogenase in the spheroplasts at 80 degrees C was 8.5 min . A spectrophotometric technique was developed for determining the membrane-bound hydrogenase in the presence of sulfhydryl compounds with methylene blue as electron acceptor . The maximal specific activity of hydrogenase in extracts prepared in the anaerobic conditions in the presence of dithiothreitol and Mg2+ and Mn2+ ions was 10 +/- 3 units per 1 mg of protein, which closely corresponded with the activity of hydrogenase in the whole cells . Almost all activity of hydrogenase assayed with methylene blue was localized in the membrane fraction . The activity of soluble NAD-specific hydrogenase was not detected . Large particles located in 60-70% sucrose had the highest hydrogenase activity upon fractionation in a continuous sucrose concentration gradient . The second, lower peak of the hydrogenase activity was detected in fractions of 40--50% sucrose . As was found by electron microscopy, the size of membrane vesicles with the hydrogenase activity varied within the range of 68--156 nm . The membrane preparations possessed the activity of NADH-dehydrogenase, NADH-oxidase and succinate dehydrogenase as well.

Dermatologica, 1979, 159(Suppl 1), 56 - 9
{Opportunistic fungi in the atmosphere}; Corbaz R; Our knowledge in aerobiology has considerably increased in the last 20 years due to new trapping methods . The use of volumetric spore traps permits to accurately analyze the particles suspended in the air and their evolution, while the deposition method gives an incomplete picture of the real air content . The air microflora changes considerably during 24 h: some groups of spores occur in the morning, others in the afternoon and yeast-like organisms during the night . The most common fungi in the atmosphere are Cladosporium sp., Sporodbolomyces considerably increase (20-10(3) times) the number of spores released in the atmosphere . Aspergillus flavus, thermophilic actinomycetes, Botrytis cinera and Didymella exitialis are cited as examples.

Antonie Van Leeuwenhoek, 1979, 45(4), 557 - 64
Preliminary quantification of immunological relationships among the transaldolases of the genus Bifidobacterium; Sgorbati B; The immunological relatedness among the transaldolases (dihydroxyacetone transferase, E.C . 2.2.1.2) of twenty species of the genus Bifidobacterum has been tested by the microcomplement fixation method, using B . thermophilum (B . ruminale) RU326 (= ATCC 25866), B . cuniculi RA93 (= ATCC 27916) and B . 'minimum' (DNA homology group) F392 (= ATCC 27916) as references . Based on the serological relationships of the transaldolases, expressed either as indices of dissimilarity or as immunological distances, the twenty species of the genus Bifidobacterium were arranged into clusters . These clusters generally coincided with the immunological groups obtained previously by the immunodiffusion method (Sgorbati and Scardovi, 1979).

Antonie Van Leeuwenhoek, 1979, 45(1), 129 - 40
Immunological relationships among transaldolases in the genus Bifidobacterium; Sgorbati B et al.; Antisera were prepared against electrophoretically homogeneous transaldolase (dihydroxyacetone transferase, E.C . 2.2.1.2.) of Bifidobacterium thermophilum (B . ruminale) RU326 (ATCC25866), B . cuniculi RA93 (ATCC27916) and B . 'minimum' (homology group) F392 (ATCC 27538) . Crude extracts of eighty six strains previously assigned to twenty one species of the genus Bifidobacterium on the basis of deoxyribonouclelic acid (DNA) homology (DNA-DNA hybridization), were compared by double diffusion tests on Ouchterlony plates . Eight groups of identical antigenic specificity were recognized . By analysis of the spur formation, the groups of identical specificity were arranged in preliminary sequences of decreasing similarity to each of the three homologous transaldolases used as reference points . The relationships between immunological data and the genetic similarity among the species of the genus measured by means of DNA-DNA hybridization were discussed together with some relevant points of bifidal ecology.

Acta Microbiol Pol, 1979, 28(3), 231 - 6
Aflatoxin B1 effect on beta-D-galactosidase in some starter cultures; Phutela RP et al.; Six starter cultures were examined for their ability to induce beta-D-Galactosidase activity . The maximum (713 x 10(-4) units/ml) and minimum (140 x 10(-4) units/ml) enzyme activity was shown by Lactobacillus bulgaricus and Streptomyces thermophilus, respectively . The maximum inhibition by aflatoxin B1 of induction and activity of enzyme in vitro was observed at 60 micrograms/ml in all cultures . However, low levels of the toxin stimulated induction of the enzyme in Streptococcus lactis and S . thermophilus.

Z Allg Mikrobiol, 1979, 19(2), 97 - 106
Fructose-1,6-bisphosphate aldolase from Vibrio marinus, a psychrophilic marine bacterium; Jones LP et al.; Fructose-1,6-bisphosphate aldolase (Fru-P2A) from a psychrophilic marine bacterium was found to be Class II aldolase based on activation by K+, activation by divalent cations, inactivation by EDTA, low molecular weight, and similar values for Km, Vmax, and Arrhenius activation energy . This enzyme was not markedly different in amino acid composition from the enzymes from mesophilic and thermophilic organisms, yet it has unusual thermal properties.

Biochemistry, 1978 Dec 26, 17(26), 5705 - 13
Histone-histone interactions in a lower eukaryote, Tetrahymena thermophila; Glover CV et al.; The six pairwise interactions of Tetrahymena macronuclear histones H2A (formerly HX), H2B, H3, and H4 have been studied using the techniques of circular dichroism and continuous variation . Parallel experiments have been performed with calf thymus histones as controls, and the 12 possible interspecies pairs have also been examined . The behavior of three of the ciliate histones, H2A, H2B, and H3 is virtually identical with that of their vertebrate counterparts . Tetrahymena H4 exhibits a pattern of interactions identical with that of calf thymus H4, but displays significant quantitative differences in the strength of those interactions . Although we have not entirely eliminated the possibility that these differences may be artifactual, we suggest that they reflect the unique primary structure of Tetrahymena H4 . These results extend the pattern of histone-histone interactions defined for plants, animals, and fungi to include the protists as well and imply the strong evolutionary conservatism of this pattern in spite of the electrophoretic and compositional differences which exist between homologous histones of different organisms . In addition, they demonstrate conclusively that histone HX should be redefined as Tetrahymena H2A.

Boll Soc Ital Biol Sper, 1978 Dec 15, 54(23), 2355 - 61
{5'-methylthioadenosine phosphorylase from Caldariella acidophila . 1 . Purification and partial characterization}; Oliva A et al.; 5'-Methylthioadenosine phosphorylase has been isolated from C.acidophila, a thermophilic bacterium living in acid hot springs at temperatures ranging from 63 to 89 degrees C . The enzyme has been purified to homogeneity in 32% yield . The enzyme shows a high degree of thermophilicity, its temperature optimum being 93 degrees C in the in vitro assay . The enzyme is exceptionally stable; no loss of activity was observable after exposure for 1 h at 100 degrees C . The optimum pH is about 7,2, with one-half of the maximal activity occurring at pH 6 and 9 . The apparent Km for the substrates are: 8,3 x 10(-5) M for MTA and 4,3 x 10(-4) M for phosphate ions.

J Cell Sci, 1978 Dec, 34, 303 - 19
Nuclear divisions with reduced numbers of microtubules in Tetrahymena; Jaeckel-Williams R; Ultrastructural observations on micronuclei of Tetrahymena thermophila, WH-6 dividing in the presence of colchicine suggest that the microtubules present at the time of drug addition persist, but no new microtubules are formed . It is suggested that the great amount of elongation seen in these drug-treated micronuclei may not be due to the persisting microtubules alone . On the basis of ultrastructural and light-microscopic observations, it is proposed that at least part of the motive force responsible for the elongation which occurs in colchicine may be due to an active sliding of the micronuclear membrane on some element of the cell cortex . In addition, the macronucleus of this species of Tetrahymena was found to be capable of dividing in colchicine in the absence of internal nuclear microtubules.

Eur J Biochem, 1978 Dec, 92(2), 509 - 19
Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8 . 1 . Purification and some properties of the purified factors; Arai K et al.; Polypeptide chain elongation factors have been purified from an extreme thermophile, Thermus thermophilus HB8 . By chromatography on a DEAE-Sephadex column, the factors were separated into two peaks; peak I contained a complex of EF-Tu and EF-Ts, while peak II was composed of EF-Tu.gdp and EF-G . These factors were subsequently purified to homogeneous states and crystallized . The EF-Tu . EF-Ts complex could be resolved into EF-Tu and EF-Ts by chromatography on a Sephadex G-200 column in the presence of 8 M guanidine-HCl . The complex could be reconstituted from EF-Tu and the renatured EF-Ts . No immunological cross-reaction was detected between EF-Tu, EF-Ts, and EF-G from T . thermophilus and the antibodies to their corresponding Escherichia coli factors . The molecular weight of EF-Tu . GDP determined by sedimentation equilibrium and sodium dodecylsulfate/polyacrylamide gel electrophoresis was 49000 and 51000 respectively . On the other hand, the molecular weight of EF-Ts was estimated as 27000 and 64000, respectively, by sodium dodecylsulfate/polyacrylamide gel electrophoresis and Sephadex gel filtration, suggesting that the protein existed probably as a dimer . The molecular weight of the EF-Tu . EF-Ts complex determined by sedimentation equilibrium and by gel filtration, was 142000 and 220000, respectively . Since the molar ratio of EF-Tu to EF-Ts in the EF-Tu . EF-Ts complex was one to one, it was suggested that the complex was composed of 2 mol each of EF-Tu and EF-Ts . The molecular weight of EF-G was estimated as 85000, 80000 and 78000 by equilibrium centrifugation, gel filtration, and sodium dodecylsulfate/polyacrylamide gel electrophoresis respectively.

Eur J Biochem, 1978 Dec, 92(2), 521 - 31
Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8 . 2 . Catalytic properties; Arai K et al.; Catalytic properties of the elongation factors from Thermus thermophilus HB8 have been studied and compared with those of the factors from Escherichia coli . 1 . The formation of a ternary guanine-nucleotide . EF-Tu . EF-Ts complex was demonstrated by gel filtration of the T . thermophilus EF-Tu . EF-Ts complex on a Sephadex G-150 column equilibrated with guanine nucleotide . The occurrence of this type of complex has not yet been proved with the factors from E . coli . 2 . The dissociation constants for the complexes of T . thermophilus EF-Tu . EF-Ts with GDP and GTP were 6.1 x 10(-7) M and 1.9 x 10(-6) M respectively . On the other hand, T . thermophilus EF-Tu interacted with GDP and GTP with dissociation constants of 1.1 x 10(-9) M and 5.8 x 10(-8) M respectively . This suggests that the association of EF-Ts with EF-Tu lowered the affinity of EF-Tu for GDP by a factor of about 600 and facilitated the nucleotide exchange reaction . 3 . Although the T . thermophilus EF-Tu . EF-Ts complex hardly dissociates into EF-Tu and EF-Ts, a rapid exchange was observed between free EF-Ts and the EF-Tu . EF-Ts complex using 3H-labelled EF-Ts . The exchange reaction was independent on the presence or absence of guanine nucleotides . 4 . Based on the above findings, an improved reaction mechanism for the regeneration of EF-Tu . GTP from EF-Tu . GDP is proposed . 5 . Studies on the functional interchangeability of EF-Tu and EF-Ts between T . thermophilus and E . coli has revealed that the factors function much more efficiently in the homologous than in the heterologous combination . 6 . T . thermophilus EF-Ts could bind E . coli EF-Tu to form an EF-Tu (E . coli) . EF-Ts (T . thermophilus hybrid complex . The complex was found to exist in a dimeric form indicating that the property to form a dimer is attributable to T . thermophilus EF-Ts . On the other hand, no stable complex between E . coli EF-Ts and T . thermophilus EF-Tu has been isolated . 7 . The uncoupled GTPase activity of T . thermophilus EF-G was much lower than that of E . coli EF-G . T . thermophilus EF-G formed a relatively stable binary EF-G . GDP complex, which could be isolated on a nitrocellulose membrane filter . The Kd values for EF-G . GDP and EF-G . GTP were 6.7 x 10(-7) M and 1.2 x 10(-5) M respectively . The ternary T . thermophilus EF-G . GDP . ribosome complex was again very stable and could be isolated in the absence of fusidic acid . The stability of the latter complex is probably the cause of the low uncoupled GTPase activity of T . thermophilus EF-G.

Eur J Biochem, 1978 Dec, 92(2), 533 - 43
Studies on polypeptide-chain-elongation factors from an extreme thermophile, Thermus thermophilus HB8 . 3 . Molecular properties; Nakamura S et al.; Molecular properties of the polypeptide chain elongation factors from Thermus thermophilus HB8 have been investigated and compared with those from Escherichia coli . 1 . As expected, the factors purified from T . thermophilus were exceedingly heat-stable . Even free EF-Tu not complexed with GDP was stable after heating for 5 min at 60 degrees C . 2 . GDP binding activity of T . thermophilus EF-Tu was also stable in various protein denaturants, such as 5.5 M urea, 1.5 M guanidine-HCl, and 4 M LiCl . 3 . Amino acid compositions of EF-Tu and EF-G from T . thermophilus were similar to those from E . coli . On the other hand, amino acid composition of T . thermophilus EF-Ts was considerably different from that of E . coli EF-Ts . 4 . In contrast to E . coli EF-Tu, T . thermophilus EF-Tu contained no free sulfhydryl group, but one disulfide bond . The disulfide bond was cleaved by sodium borohydride or sodium sulfite under native conditions . The heat stability of the reduced EF-Tu . GDP, as measured by GDP binding activity, did not differ from that of the untreated EF-Tu . GDP . 5 . T . thermophilus EF-Ts contained, in addition to one disulfide bond, a sulfhydryl group which could be titrated only after complete denaturation of the protein . 6 . Under native conditions one sulfhydryl group of T . thermophilus EF-G was titrated with p-chloromercuribenzoate, while the rate of reaction was very sluggish . The sulfhydryl group appears to be essential for interaction with ribosomes, whereas the ability to form a binary GDP . EF-G complex was not affected by its modification . The protein contained also one disulfide bond . 7 . Circular dichroic spectra of EF-Tu from T . thermophilus and E . coli were very similar . Binding of GDP or GTP caused a similar spectral change in both . T . thermophilus and E . coli EF-Tu . On the other hand, the spectra of T . thermophilus EF-G and E . coli EF-G were significantly different, the content of ordered structure being higher in the former as compared to the latter.

Biochim Biophys Acta, 1978 Nov 21, 521(1), 288 - 94
An enhanced thermostability in thermophilic 5-S ribonucleic acids under physiological salt conditions; Nazar RN et al.; The secondary structure of 5-S rRNAs of Thermus aquaticus (an extreme thermophile), Bacillus stearothermophilus (a moderate thermophile) and Escherichia coli (a mesophile) was compared using thermal denaturation techniques under varying ionic conditions . At a low ionic strength (10 mM K+), the Tm of T . aquaticus 5-S RNA differed by only 1 degrees C from that of E . coli RNA and the molecule was fully denatured well below the optimum growth temperature of the thermophile . The internal Na+, K+ and Mg2+ concentrations of T . aquaticus cells were determined to be 91 mM, 130 mM and 59 mM, respectively . Under these salt conditions, T . aquaticus 5-S RNA was significantly more stable than E . coli RNA and the 5-S RNA from B . stearothermophilus was intermediate as is its optimum growth temperature . The results suggest that the thermostability of macromolecules from thermophilic organisms may be specially dependent on the internal salt concentration . Furthermore, under these salt conditions, most of the secondary structure of the RNA remained stable at the optimum growth temperatures suggesting that ribosomal RNAs of thermophilic organisms contribute more to the thermostability of the ribosome than previously thought.

Parazitologiia, 1978 Nov-Dec, 12(6), 529 - 38
{Ectoparasites of small mammals in the eastern portion of the Baikal-Amure mainline}; Volkov VI et al.; 11.501 ectoparasites of 45 species were collected from small mammals (rodents, shrews and double-toothed rodents) in the north-eastern part of Priamurje . The parasitecoenoses are formed in general by gamasid mites (60.4%) and lice(24.7%) but faunistically fleas (19 species or 42.2%) are dominant . Among all ectoparasites the gamasid mites, Laelaps clethrionomydis and Hirstionyssus isabellinus, and the louse Hoplopleura acanthopus account for 64.9% . The impoverishment of the arthropod fauna in the northern part of Priamurje as compared to southern ones proceeds at the expence of chigger mites and more thermophilous species of gamasids and ixodids . With cultivation of forests the content of members of parasitecoenoses becomes more homogenous and their abundance falls.

J Biochem (Tokyo), 1978 Nov, 84(5), 1319 - 21
An isochizomer of TaqI from Thermus thermophilus HB8; Sato S et al.; A site-specific endonuclease has been isolated from Thermus thermophilus HB8 and named TthHB8I . It recognizes the same sequences as TaqI from Thermus aquaticus YT-1 does . The amount of TthHB8I in the cells was comparable to that of TaqI . T . thermophilus HB8 has an advantage over T . aquaticus YT-1 for preparation of a TaqI-like enzyme since it is easier to obtain T . thermophilus HB8 cells in quantity.

Infect Immun, 1978 Nov, 22(2), 568 - 74
Complement activation by cell wall fractions of Micropolyspora faeni; Smith SM et al.; The ability of several cell wall fractions of Micropolyspora faeni, a thermophilic actinomycete associated with farmer's lung disease, to activate complement is reported . Cell walls, obtained by mechanical disruption, were purified by enzyme treatment and chemical extractions . Fractions containing the most purified cell walls were most active in consuming complement, as measured by reduction of hemolytic complement levels of normal human serum . Cell wall fractions activated the alternative complement pathway, as shown by monitoring the conversion of C3 proactivator (factor B) to C3 activator (activated factor B) in the presence of specific cation chelators . Selective degradation of cell walls by lysozyme resulted in a decreased ability to consume complement and implicated peptidoglycan as the major complement-reactive component . The role of this nonspecific complement activation in relation to farmer's lung disease is discussed.

J Bacteriol, 1978 Nov, 136(2), 815 - 7
Thermal adaptation in yeast: obligate psychrophiles are obligate aerobes, and obligate thermophiles are facultative anaerobes; Watson K et al.; The obligate psychrophilic yeasts Torulopsis psychrophila, T . austromarina, Leucosporidium frigidum, L . gelidum, and L . nivalis were obligate aerobes and were unable to grow anaerobically . In contrast, the obligate thermophilic yeasts T . bovina, T . pintolopesii, Candida slooffii, and Saccharomyces telluris were facultative anaerobes.

Hoppe Seylers Z Physiol Chem, 1978 Nov, 359(11), 1491 - 507
The complete amino acid sequence of both subunits of C-phycocyanin from the cyanobacterium Mastigocladus laminosus; Frank G et al.; The amino acid sequences of both subunits of the C-phycocyanin from the thermophilic cyanobacterium Mastigocladus laminosus have been determined . The alpha-chain consists of 162 amino acid residues and has a molecular weight of 18000, whereas the beta-chain consists of 172 residues and has a molecular weight of 19400 . For the first three quarters of their length the polypeptide chains are 31% homologous, whereas there is no significant homology in the final quarter up to the C-terminus . This could mean that the introduction of an additional chromophore binding site in the last quarter of the beta-chain during evolution was achieved via a large number of point mutations or by exchange of the whole C-terminal part in an ancestral gene.

Vet Rec, 1978 Oct 21, 103(17), 373 - 5
The destruction of pig helminth ova and larvae in a slurry treatment process; Burden DJ et al.; It was established that a thermophilic system for the treatment of pig slurry at 55 degrees C rapidly killed the free living stages of three common pig parasites . This treatment could be beneficially incorporated in any pig slurry recycling process, whether to land or to animals.

Chromosoma, 1978 Oct 20, 69(1), 1 - 19
Life cycle variation and regulation of macronuclear DNA content in Tetrahymena thermophila; Doerder FP et al.; The mean DNA content of G2 macronuclei varies during the life cycle of the ciliate Tetrahymena thermophila . Early in the life cycle the mean is about 130 C; later it is about 94 C . In hybrids between strains A and B the decrease from 130 C to 94 C usually began after 60 fissions after conjugation . In B X B clones the decrease was complete by 50 fissions . The data suggest that there may be a genetic difference between strains A and B with respect to the onset of the decrease in DNA content . The downward regulation of the mean DNA content appears to be related to the mechanism which removes the variance in macronuclear DNA content which is added to macronuclei by unequal macronuclear division . Unequal macronuclear division regularly occurs at all stages of the life cycle, with larger macronuclei tending to divide more unequally . In the absence of regulation, unequal macronuclear division would constantly add variance to G1 macronuclei and their range would continue to increase . Analysis of the variances of G1 and G2 macronuclei suggests that at all stages of the life cycle the added variance is removed by acting upon nuclei which become too small or too large . According to this model, macronuclei with smaller amounts of DNA are regulated upward by an extra macronuclear S phase, while larger amounts are regulated downward by chromatin extrusion and the skipping of macronuclear S . The mean DNA content appears to change during the life cycle because the thresholds at which macronuclei become too small or too large are readjusted . It is postulated that these thresholds are a function of gene dosage.

Science, 1978 Oct 13, 202(4364), 219 - 21
Physiologically important stabilization of DNA by a prokaryotic histone-like protein; Stein DB et al.; The thermophilic mycoplasma Thermoplasma acidophilum has tightly bound to its DNA a protein that closely resembles the histones of eukaryotes . DNA associated with this protein is more stable than free DNA against thermal denaturation by about 40 degrees C, as shown in both native nucleoprotein and in hybrid nucleoprotein reconstituted in vitro with calf DNA . Since only about 20 percent of the DNA in this organism is associated with the histone-like protein, we suggest that its physiological function is to prevent complete separation of the DNA strands during brief exposures of the organism to denaturing conditions, and thus to facilitate rapid renaturation when normal environmental conditions return.

Biochim Biophys Acta, 1978 Oct 12, 526(2), 457 - 67
Alkaline phosphatase from Bacillus licheniformis . Solubility dependent on magnesium, purification and characterization; Schaffel SD et al.; The membrane-associated alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) from Bacillus licheniformis MC14, a facultative thermophile, was purified to homogeneity in buffer containing 0.2 M Mg2+ . The alkaline phosphatase purified in this manner is insoluble upon removal of the magnesium by dialysis . This insoluble alkaline phosphatase has been characterized and compared to the previously purified heat-solubilized enzyme (Hulett-Cowling, F.M . and Campbell, L.L . (1971) Biochemistry 10, 1364--1371).

Arch Microbiol, 1978 Oct 4, 119(1), 17 - 24
Calcium uptake and survival of Bacillus stearothermophilus; Stahl S; The calcium transport in resting vegetative cells of Bacillus stearothermophilus was studied by determining the retention of 45Ca in a membrane filter assay . The kinetics of death by vegetative cells, when suspended in buffer at 55 degrees C, was also investigated . The calcium influx required the presence of an energy source, e.g . glucose-1-phosphate and the system exhibited saturation kinetics . The requirements for survival of the thermophilic cells reflected those of the calcium transport system . Thus, cells treated with nitrogen gas showed an increased thermal stability and a decreased efflux of calcium . The initial velocity of calcium influx correlated linearly with the survival of the cells after 1 min heating at 55 degrees C . Lanthanum inhibited calcium influx and reduced survival . Magnesium did not inhibit calcium influx and could replace calcium as a stabilizing agent . The results suggest that the thermophilic cells are not intrinsically heat stable but survive due to a high cellular concentration of divalent ions.

Nucleic Acids Res, 1978 Oct, 5(10), 3457 - 67
A specific endonuclease from Bacillus caldolyticus; Bingham AH et al.; The purification and characterization of a new restriction endonuclease, BclI from the extreme thermophile Bacillus caldolyticus is reported . This enzyme recognizes the sequence : formula: (see text) and cleaves at the positions indicated by the arrows.

J Cell Sci, 1978 Oct, 33, 227 - 34
Directionality of microtubule assembly: an in vivo study with the ciliate Tetrahymena; Ng SF; A temperature-sensitive mutant homozygous for the recessive gene molb in Tetrahymena thermophila offers opportunity for studying the direction of microtubule assembly in vivo . At 39 degrees C the mutant fails to divide properly; the 2 daughter animals remain attached and bend over each other . As revealed by protargol staining, the bending results in acute turning and breaking of some of the longitudinal microtubular bands close and parallel to the surface . Hence, 2 broken microtubular ends are available for study of the problem of directionality of microtubule assembly, by assessing which of the 2 ends regenerates . In most cases the posterior portion of the longitudinal microtubular band regenerates . The present study hence supports the conclusion based on in vitro observation in other systems that microtubule assembly is predominantly unidirectional.

J Bacteriol, 1978 Oct, 136(1), 19 - 23
ATP hydrolysis and synthesis by the membrane-bound ATP synthetase complex of Methanobacterium thermoautotrophicum; Doddema HJ et al.; The membrane-bound ATP synthetase complex of Methanobacterium thermoautotrophicum showed maximum activity for ATP hydrolysis at pH 8, at temperatures between 65 and 70 degrees C, and at an ATP-Mg2+ ratio of 0.5 . Anaerobic conditions were not prerequisite for enzyme activity . The enzyme showed a Km value for ATP of 2 mM, and activity was Mg2+ dependent; Mn2+, Co2+, Ca2+, and Zn2+ could replace Mg2+ to some extent . Other nucleoside triphosphates could be hydrolyzed . N,N'-dicyclohexylcarbodiimide inhibited ATP hydrolysis . A proton-motive force, artificially imposed by a pH shift or valinomycin, resulted in ATP synthesis in whole cells . The ATP synthetase complex of the thermophilic methanogenic bacterium is similar to those described in aerobic and anaerobic microorganisms.

Science, 1978 Sep 29, 201(4362), 1187 - 91
Electrostatic effects in proteins; Perutz MF; Electrostatic effects dominate many aspects of protein behavior . When polypeptide chains fold up, most polar side chains seek the exterior, where they can be solvated . Water bound in the interior has been found between the domains of enzymes of the chymotrypsin family, and between the subunits of hemoglobin and tobacco mosaic virus protein . Assembly of this protein from disk to virus is triggered by electrostatic interactions between neighboring subunits . Lysozyme stabilizes the constellation of charges involved in the transition state of its substrate by both permanent and induced dipoles . All factors that lower the oxygen affinity of hemoglobin act by strengthening the salt bridges that constrain its quaternary deoxy (T) structure . Enzymes of thermophile bacteria owe their extra stability mostly to additional salt bridges . The rate of denaturation of hemoglobins by alkali is determined by the ionization of internal side chains with pK's of about 12.

J Allergy Clin Immunol, 1978 Sep, 62(3), 185 - 9
Farmer's lung disease among farmers with precipitating antibodies to the thermophilic actinomycetes: a clinical and immunologic study; Marx JJ et al.; Farmers were evaluated for the presence of farmer's lung disease by serologic methods and by clinical histories . From a large farming population screened serologically, 40 of 92 farmers with precipitating antibodies to the thermophilic actinomycetes consented to be evaluated for clinical evidence of farmer's lung disease . Each subject completed a standardized questionnaire which was reviewd by a trained observer . On the basis of the questionnaire and an indepth clinical history, the subjects were grouped into those farmers who had a history of farmer's lung disease (38%), those with no history (50%), or a doubtful group (12%) . When these groups were compared for radiologic changes, pulmonary function abnormalities, and immune function, no differences were discernible . None of the parameters tested were useful in predicting which farmer would develop clinical farmer's lung disease . The parameters tested did not provide reliable criteria for differentiating those patients who present without acute symptoms but relate a history of recurrent pulmonary illnesses associated with moldy forage.

Proc Natl Acad Sci U S A, 1978 Sep, 75(9), 4219 - 23
Resolution of the membrane moiety of the H+-ATPase complex into two kinds of subunits; Sone N et al.; The H+-translocating ATPase complex from the thermophilic bacterium PS3 (TF0-F1) is composed of a water-soluble part with ATP-hydrolyzing activity (TF1) and a membrane moiety with H+-conducting activity (TF0) . TF0 was obtained by treating TF0-F1 with urea and removing contaminations on a carboxymethyl-cellulose column . This TF0 contained only two kinds of subunits, band 6 protein (13,500 daltons) and band 8 protein (5400 daltons), and it was active in H+ conduction and TF1 binding when reconstituted into proteoliposomes (TF0 vesicles) . The binding of TF1 to TF0 present in vesicles restored energy-transducing activities, such as ATP-32Pi exchange, dicyclohexylcarbodiimide-sensitive ATPase, and ATP-dependent enhancement of 8-anilinonaphthalene-1-sulfonate fluorescence . Treatments such as protease digestion and chemical modification with acetic anhydride, succinic anhydride, or diazobenzenesulfonic acid destroyed the TF1-binding activity, which was caused by band 6 protein . Band 8 protein was a proteolipid that reacted specifically with dicylcohexyl-carbodiimide and seemed to play a central role in H+ conduction through the membrane.

Mikrobiologiia, 1978 Sep-Oct, 47(5), 815 - 22
{Sulfobacillus, a new genus of thermophilic sporulating bacteria}; Golovacheva RS et al.; An aerobic facultative thermophilic bacterium was isolated from the ore of the Nikolaev copper-zinc-pyrite deposit in the Eastern Kazakhstan . The bacterium is similar to Thiobacillus ferooxidans in the ability to use various energy subsrates (Fe2+, sulphide minerals and So) and in the acidophilic properties, but differs from it in the thermophilic nature, the structure of a cell wall, the ability to produce spores, and a lower GC content in DNA . Consequently, the organism has been identified as a new species of a new genus, Sulfobacillus thermosulfidooxidans gen . nov., sp . nov . The organism is presumed to play an active role in oxidation of sulphide ores and their warming in deposits.

Vet Med (Praha), 1978 Sep, 23(9), 569 - 72
{Detection of inhibitory substances in milk using the modified reduction micromethod}; Konecny S; In order to rationalize work and to reduce the consumption of laboratory glassware a reduction micromethod with the bacterial stain Streptococcus thermophilus has been modified; the method is used to detect inhibitory substances in genuine, pausteurized and dried milk, and it is performed on a serological plate made from organic glass . The method is sensitive to most antibiotics used in the veterinary clinical practice . The detection sensitivity to penicillin residua ranges from 0.0015 to 0.0015 i.u . in 1 ml of milk . The method may be assessed as semi-quantitative.

J Bacteriol, 1978 Sep, 135(3), 1043 - 52
Lipid and protein composition of membranes of Bacillus megaterium variants in the temperature range 5 to 70 degrees C; Rilfors L et al.; Membranes were prepared from four temperature range variants of Bacillus megaterium: one obligate thermophile, one facultative thermophile, one mesophile, and one facultative psychrophile, covering the temperature interval between 5 and 70 degrees C . The following changes in membrane composition were apparent with increasing growth temperatures: (i) the relative amount of iso fatty acids increased and that of anteiso acids decreased, the ratio of iso acids to anteiso acids being 0.34 at 5 degrees C and 3.95 at 70 degrees C, and the pair iso/anteiso acids thus seemed to parallel the pair saturated/unsaturated acids in their ability to regulate membrane fluidity; (ii) the relative/unsaturated acids in their ability to regulate membrane fluidity; (ii) the relative amount of long-chain acids (C16 to C18) increased fivefold over that of short-chain acids (C14 and C15) between 5 and 70 degrees C; (iii) the relative amount of phosphatidylethanolamine increased, and this phospholipid accordingly dominated in the thermophilic strains, whereas diphosphatidylglycerol was predominant in the two other strains; and (iv) the ratio of micromoles of phospholipid to milligrams of membrane protein increased three-fold between 5 and 70 degrees C . Moreover, a quantitative variation in membrane proteins was evident between the different strains . Briefly, membrane phospholipids with higher melting points and packing densities appeared to be synthesized at elevated growth temperatures.

Mikrobiologiia, 1978 Sep-Oct, 47(5), 947 - 52
{Acidophilic obligate thermophilic bacteria, Bacillus acidocal-darius, isolated from the hot springs and soil of Kunashir Island}; Loginova LG et al.; Obligate thermophilic acidophilic cultures of Bacillus acidocaldarius were isolated from hot springs and soil of the Kunashir Island (the Kuril Islands) and Southern Sakhalin; their optimum temperature of growth is 65 degrees C and pH is 3.3--4.0 . It has been shown for the first time that these organisms can be isolated from springs with either acidic or neutral and weakly alkaline pH values.

Z Med Lab Diagn, 1978 Aug, 19(4), 251 - 6
{Epidemiological study on the demonstration of precipitating antibodies against Micropolyspora faeni in agricultural workers}; Thiele E et al.; The authors report on the frequency (as determined by the double diffusion test according to Ouchterlony) of circulating precipitating antibodies against thermophil actinomycetes of the genus Micropolyspora faeni in 1666 blood samples from 1376 agricultural workers . These workers belonged to the main professional categories in two districts of the German Democratic Republic and were examined by members of the Research Group on Occupational Medicine in Agriculture of the Ernst Moritz Arndt University during the years 1972-1975 . 290 workers were examined twice or three times in a longitudinal study . Positive serological findings were obtained from 5.1% of the subjects under investigation . There were no statistically significant differences with regard to occupational category, sex, age, and season . Nevertheless, there were marked regional differences . The results obtained show that there is no need for special medical supervision of the agricultural workers with regard to thoracic actinomycosis (farmers' lung) . Further examinations are of course necessary in case of suspected or clinically manifest affection.

Genetics, 1978 Aug, 89(4), 703 - 10
Induced resistance to 6-methylpurine and cycloheximide in Tetrahymena . II . Variation within vegetative cultures of micronucleate T . thermophila and amicronucleate T . pyriformis; Byrne BC; Resistance to 6-methylpurine and cycloheximide has been induced in both the micronucleate species Tetrahymena thermophila and the amicronucleate species T . pyriformis . Resistance follows only after mutagen treatment and vegetative growth . The frequencies with which resistant variants are induced and the independence of mutagenesis and selection are demonstrated . All evidence is consistent with the hypothesis that macronuclear subunits are assorting in both species during vegetative growth to produce new phenotypes among subclones.

Genetics, 1978 Aug, 89(4), 695 - 702
Induced resistance to 6-methylpurine and cycloheximide in tetrahymena . I . Germ line mutants of T . thermophila; Byrne BC et al.; Two new mutant lines of Tetrahymena thermophila (T . pyriformis, syngen 1), each conferring resistance to a different agent, are described . Resistance to cycloheximide and 6-methylpurine are each determined by dominant genes, ChxA2 and Mpr; the traits show phenotypic assortment . The method used to select these mutations, the critical importance of backcrossing to wild type following mutagenesis, and the utility of these marker genes in further mutagenic selection schemes and studies of the sexual cycle of Tetrahymena are noted.

Zentralbl Bakteriol {B}, 1978 Aug, 167(1-2), 171 - 6
Occurrence and thermoresistance of thermophilic streptomycetes spores in milk; Falkowski J; 200 samples of raw milk and 100 samples of pasteurized milk were tested for the presence of thermophilic streptomycetes . One ml of raw and pasteurized milk contained mean 8.42 and 14.95 thermophilic streptomycetes, respectively . The isolated strains belonging to the species Thermoactinomyces vulgaris Tsiklinsky 1899 and Micromonospore sp . (Agre et al., 1972) were identified . To determine the survival of spores of these thermophiles after pasteurization, samples of milk were heated at 63.5 degrees C/30 min and at 75 degrees C/15 s . Ampouls with this milk, dairy pasteurized milk and raw milk were heated for the same time intervals at 100, 105, 110 and 120 degrees C . Thermophilic streptomycetes spores contained in the investigated milk samples survived the both of pasteurization temperatures . A relative high number of thermophilic streptomycetes spores survive the heat treatment at 100 degrees C . The heat resistance at 105, 110 and 120 degrees C is distinctly lower . The most resistant in the range of the testet temperatures proved spores of the species Thermoactinomyces vulgaris Tsiklinsky 1899.

Zentralbl Bakteriol {B}, 1978 Aug, 167(1-2), 165 - 70
{The thermophilic streptomycetes flora in milk powders and condensed milk products (author's transl)}; Falkowski J; 247 specimens of powdered milk and 165 of condensed milk were tested for their contamination with thermophile Streptomycetes . Colonies of these contaminants were isolated from all specimens of powdered milk and from 73 samples of condensed sweeted milk . The isolated strains corresponded with the following species of thermophilic Streptomycetes: Thermoactinomyces vulgaris, Tsiklinsky 1899, Thermoactinomyces vulgaris, Tsiklinsky 1899 "giant colonies", Micromonospora sp . (Agre et al., 1).

J Bacteriol, 1978 Aug, 135(2), 334 - 41
Structural basis of the thermostability of monomeric malate synthase from a thermophilic Bacillus; Chell RM et al.; Malate synthases from a thermophilic Bacillus and Escherichia coli have been isolated in a high state of purity . Molecular weights of these two proteins determined in the native state and after denaturation in sodium dodecyl sulfate-mercaptoethanol show that the enzymes are monomeric . This conclusion is supported, for the thermophile enzyme, by the result of an electrophoretic analysis of that protein after treatment with dimethylsuberimidate and denaturation . The thermophilic Bacillus malate synthase is considerably more thermostable than its mesophilic counterparts from E . coli, Bacillus licheniformis, and Pseudomonas indigofera . It is, however, markedly labilized by an increase in the ionic strength of the medium brought about by the addition of 0.2 M potassium chloride or in pH above 9 . Increased ionic strength has little effect on the thermostability of the mesophilic bacterial malate synthases . These observations provide strong support for the idea that monomeric proteins in thermophiles owe their unusual heat stability to the presence of salt bridges in their tertiary structure.

J Biol Chem, 1978 Jul 10, 253(13), 4599 - 603
The protonmotive force and beta-galactoside transport in Bacillus acidocaldarius; Krulwich TA et al.; The acidophilic and thermophilic bacterium, Bacillus acidocaldarius maintains a cytoplasmic pH between 5.85 and 6.31 over a range of external pH from 2.0 to 4.5 . Consistently, the pH optimum of beta-galactosidase, as assayed in cell extracts, is between pH 6.0 and 6.5 . An electrical potential (delta-psi), interior positive, is also maintained across the membrane . A delta-psi of approximately 34 mV was calculated from determinations of thiocyanate uptake by cells at pH 3.5 . Addition of the proton conductor carbonyl cyanide m-chlorophenylhydrazone increased the delta-psi . Treatment of cells with valinomycin (in the absence of external potassium ions) or high concentrations of thiocyanate, to abolish the delta psi, resulted in collapse of the transmembrane proton gradient (delta pH) . Active transport of methylthio-beta, D-galactoside occurred optimally at pH 3.5 . Transport of the galactoside was inhibited by various compounds which could dissipate the transmembrane delta pH and by respiratory inhibitors . A decrease in the delta pH and an increase in the delta psi occurred upon addition of methylthio-beta, D-galactoside to cells of B . acidocaldarius . Thus the transport of this solute appears to involve an electrogenic symport with protons . The transport system is most active at 50 degrees C and shows little activity at 25 degrees C, although the delta pH is the same at the two temperatures . Gramicidin inhibits methylthio-beta, D-galactoside transport equally effectively at 50 degrees C and 25 degrees C, while nigericin inhibits only after a lag at 25 degrees C.

Biochim Biophys Acta, 1978 Jul 7, 525(1), 162 - 70
Purification and properties of trehalase from the thermophilic fungus Humicola lanuginosa; Prasad AR et al.; Trehalase (alpha,alpha-Trehalose glucohydrolase, EC 3.2.1.28) was partially solubilized from the thermophilic fungus Humicola lanuginosa RM-B, and purified 184-fold . The purified enzyme was optimally active at 50 degrees C in acetate buffer at pH 5.5 . It was highly specific for alpha,alpha-trehalose and had an apparent Km = 0.4 mM at 50 degrees C . None of the other disaccharides tested either inhibited or activated the enzyme . The molecular weight of the enzyme was around 170 000 . Trehalase from mycelium grown at 40 and 50 degrees C had similar properties . The purified enzyme, in contrast to that in the crude-cell free extract, was less stable . At low concentration, purified trehalase was afforded protection against heat-inactivation by "protection against heat-inactivation by "protective factor(s)" present in mycelial extracts . The "protective factor(s)" was sensitive to proteolytic digestion . It was not diffusible and was stable to boiling for at least 30 min . Bovine serum albumin and casein also protected the enzyme from heat-inactivation.

Mikrobiologiia, 1978 Jul-Aug, 47(4), 653 - 8
{Variability of the proteolytic activity in the thermotolerant actinomycete, Thermoactinomyces vulgaris}; Loginova LG et al.; The thermophilic culture of Thermoactinomyces vulgaris PA-11-4a, like other actinomycetes, is characterized by a high variability . This strain produces protease . The property is maintained if spores are incubated in sterilized distilled water for 30 min after lyophilization and then inoculated onto a solid medium (Petri plates) in order to select small dark-coloured colonies . The activity of protease does not decrease if such colonies are grown on solid media during seven months at 4 degrees C.

Arch Microbiol, 1978 Jul, 118(1), 49 - 53
Stimulation of growth and glucose catabolite enzymes by succinate in some thermophilic fungi; Wali AS et al.; Thermophilic Humicola lanuginosa, Penicillium duponti, Sporotrichum thermophile and Mucor pusillus required succinate in addition to glucose for optimal growth . The requirement for succinate was concentration-dependent and the concentration needed for one half of the maximal growth was 6.14mM . In the presence of succinate, glucose utilization from the medium was markedly increased and this was associated with increased levels of the enzymes of the glycolytic and Krebs cycle pathways . Addition of succinate to cultures growing in glucose at any stage of growth stimulated the growth with the resulting rate of growth remaining high if the addition was made within 3 days of inoculation . Cycloheximide (71.4 micrometer) prevented the succinate-mediated derepression of the enzymes suggesting that succinate may remove the catabolite repression in the presence of glucose.

Biochem J, 1978 Jul 1, 173(1), 165 - 77
Isolation and characterization of isocitrate lyase from a thermophilic Bacillus sp; Chell RM et al.; Isocitrate lyase was isolated in homogeneous state from a thermophilic Bacillus . The enzyme has a mol.wt . of 180000 and a pI of 4.5 and contains threonine as the N-terminal residue . It resembles in size the cognate enzyme from the mesophilic bacterium Pseudomonas indigofera, but is smaller than the enzyme from the eukaryotic fungus Neurospora crassa . All three lyases are tetramers and similar in amino acid composition, but the thermophile enzyme is distinctive from its mesophilic coutnerparts in possessing a lower catalytic-centre activity, greater resistance to chemical and thermal denaturation and fewer thiol groups and in being strongly activated by salts . Salt activation, by 0.4M-KCl, is about 3-fold at 30 degrees C and pH 6.8 and weakens progressively as the temperature or pH is raised . The activation is probably due to a change in the enzyme conformation caused by the electrolyte modifying the interaction between charged groups or between hydrophobic groups in protein . The possible significance of the salt activation, of the relative paucity of thiol groups and of the greater resistance to chemical denaturants is discussed . Besides its effect on the Vmax., KCl produces large increases in the magnitude of several kinetic parameters . A rise in reaction temperature from 30 to 55 degrees C produces a somewhat similar result . In view of these peculiar features, the patterns of inhibition of enzyme activity by compounds such as succinate and phosphoenolpyruvate were examined at 30 and 55 degrees C in the presence and absence of KCl.

Can J Microbiol, 1978 Jul, 24(7), 794 - 7
Isolation anc characterization of actinophages of Thermoactinomyces and Micropolyspora; Kurup VP et al.; Two new actionphages infecting thermophilic actinomycetes were isolated from lysogenic cultures of Thermoactinomyces candidus and Micropolyspora faeni . The physiochemical properties, morphology, host range, electron microscopy, and immunology of the two phages, phi-115A (ATCC 29680-B1) and phi-150A (ATCC 29681-B1), are reported.

Proc Natl Acad Sci U S A, 1978 Jul, 75(7), 3158 - 62
Cycloheximide resistance can be mediated through either ribosomal subunit; Sutton CA et al.; Two cycloheximide-resistant mutants of Tetrahymena thermophila were analyzed to determine the site of their cycloheximide resistance . The mutations in both strains had been previously shown to be genetically dominant and located at separate loci (denoted Chx-A and Chx-B) . Strains carrying these mutations were readily distinguished by the extent to which they were resistant to the drug . The homozygous double mutant was more resistant than either single mutant . Cell-free extracts of wild type and of the three mutant strains, assayed for protein synthetic activity by both runoff of natural mRNA and poly(U)-dependent phenylalanine polymerization, demonstrated that in vitro the mutants were all more resistant than the wild type . Further fractionation of the cell-free systems into ribosomes and supernates localized cycloheximide resistance to the ribosome for both Chx-A and Chx-B homozygotes . Ribosome dissociation and pairwise subunit mixing in the in vitro system indicated that ribosome resistance was conferred by the 60S subunit from one strain whereas resistance in the other strain was mediated through the 40S subunit . This was further confirmed by reconstruction of all four cycloheximide-resistance "phenotypes" by mixing ribosomal subunits from appropriate strains . This finding suggests that the mechanisms by which these mutations confer resistance to cycloheximide are different.

J Biochem (Tokyo), 1978 Jul, 84(1), 193 - 203
Purification and properties of acetate kinase from Bacillus stearothermophilus; Nakajima H et al.; 1 . Acetate kinase {EC 2.7.2.1} from an thermophile, B . stearothermophilus, was purified and crystalized . 2 . This enzyme was shown to be a tetramer of identical subunits which had a molecular weight of about 40,000 . Amino acid analysis showed no SH group . By analyzing the CD spectrum it was deduced that this enzyme is composed of 36% beta-structure, 21% alpha-helix and 43% unordered structure . 3 . This enzyme shared many common enzymatic properties with the counterpart from mesophiles, i.e . pH optimum, substrate specificity, requirement of metal ions and essential amino acid residues necessary for the catalytic activity . However, this enzyme was remarkably thermostable . 4 . A plot of the reaction velocity against the concentration of acetate, ADP or acetyl phosphate gave a curve of the Michaelis-Menten type . However, such a plot against ATP gave a sigmoid curve, suggesting a homotropic allosteric nature of the enzyme . 5 . From the results of chemical modification it was deduced that an amino group and an imidazole group, at least, are involved in the active site of the enzyme.

Nucleic Acids Res, 1978 Jun, 5(6), 1729 - 39
A restriction enzyme Tha I from the thermophilic mycoplasma Thermoplasma acidophilum; McConnell DJ et al.; A type II restriction enzyme (Tha I) has been isolated from the thermophilic mycoplasma Thermoplasma acidophilum . A new method of general application was used to determine the DNA sequence cleaved by the enzyme . Tha I cuts DNA in the centre of the sequence CGCG . Single-stranded DNA is not a substrate . Tha I does not cut T . acidophilum DNA which is presumably modified . This is the first description of a restriction enzyme from a mycoplasma . Because Tha I is easily prepared in large amounts of approximately 10(5) units per gram of cells, it will be a valuable addition to the battery of restriction enzymes used in studies of DNA sequences . It is active at high temperatures and may therefore be useful for special purposes requiring more extreme conditions.

J Biochem (Tokyo), 1978 Jun, 83(6), 1693 - 8
Membrane properties of an extreme thermophile . II . Membrane functions underlying leucine transport and their relation with thermotropic phase transitions; Wakayama N; Various membrane properties of Thermus thermophilus HB8 were studied in order to elucidate the mechanism and implication of cell adaptation to a high temperature environment . 1 . The profile of temperature dependence of the amino acid uptake rates was similar for the membrane obtained from cells grown at both 61 degrees C and 77 degrees C . 2 . The remaining uptake activity after heat treatment paralleled the membrane potential generating ability, as measured by auramine O fluorescence response to the respiration substrate, and was not dependent on the growth temperature . 3 . The temperature dependence of the efflux of the accumulated amino acid reflects the physical phase of the membrane and is dependent on the growth temperature.

J Biochem (Tokyo), 1978 Jun, 83(6), 1687 - 92
Membrane properties of an extreme thermophile . I . Detection of the phase transition and its dependence on growth temperature; Wakayama N et al.; Phase transition was detected by a fluorescence polarization technique in the membrane of Thermus thermophilus HB8: it was found to be a function of cell growth temperature when the cell growth temperature was varied between 50-80 degrees C . A systematic relation between the phase transition temperature and the growth temperature was observed . Differential scanning calorimetry was also applied . The phase transition was found to take place between 34 and 55 degrees C for cells grown at 50 degrees C and between 52 and 80 degrees C for cells grown at 80 degrees C.

J Biochem (Tokyo), 1978 Jun, 83(6), 1521 - 32
Purification and properties of an extracellular exonuclease from Thermus thermophilus HB8; Takahashi M et al.; An extracellular exonuclease has been found and purified about 10,000-fold from the culture broth of an extreme thermophile, Thermus thermophilus HB8 . The enzyme had an isoelectric point at pH 5.1 and seems to be of a multimolecular type, with molecular weights estimated to be ca . 530,000 (Peak I) and around 330,000 (Peak II) by gel filtration . The properties of the most highly purified enzyme fraction, Peak I were investigated . The enzyme requires divalent cations (Mg2+ greater than Sn2+ greater than Ca2+, Mn2+) and is inactive in the presence of EDTA . The pH optimum is 9.4-9.5 in glycine-NaOH buffer and the optimum temperature is 85 degrees C . The rate of hydrolysis increases in the order heat-denatured DNA greater than native DNA greater than RNA . The enzyme hydrolyzes deoxyoligonucleotides bearing 5'-monophosphate to liberate 5'-mononucleotides in an exonucleolytic manner . However, oligonucleotides lacking a 5'-phosphoryl group, irrespective of the presence or absence of phosphate at the 3'-termini, give both 5'-mononucleotides and dinucleoside monophosphates derived from the 5'-termini . It was also found that dinucleotides terminated with a 5'-phosphoryl group were cleaved to 5'-mononucleotides, but dinucleoside monophosphates were resistant to this enzyme . This exonuclease should be useful in the direct determination of sequence at the 5'-terminus and the penultimate position of oligonucleotides by the use of high-performance liquid chromatography.

Ann Allergy, 1978 Jun, 40(6), 385 - 6
On respirable organic antigens, mattresses and some idiopathic lung diseases; Schwartz HJ et al.; In surveying the environment of workers in a mattress factory thermophilic actinomycetes as well as phycomycetes and aspergillus species were isolated readily from work room dust, finished bat, sissal and the interior of baled linters . Those persons working with cotton products were shown to have a heavy, daily exposure to organisms known to be implicated in the pathogenesis of hypersensitivity pneumonitis . The authors suggest that further study of the inhaled antigens to which cotton workers are exposed is indicated before discarding immunologic mechanisms to explain the development of byssinosis and other pulmonary disorders to which such workers are subject.

Appl Environ Microbiol, 1978 Jun, 35(6), 1019 - 26
Thermophilic methanogenesis in a hot-spring algal-bacterial mat (71 to 30 degrees C); Ward DM; Algal-bacterial mats which grow in the effluent channels of alkaline hot springs provided an environment suitable for studying natural thermophilic methane producing bacteria . Methane was rapidly produced in cores taken from the meat and appeared to be an end product of decomposition of the algal-bacterial organic matter . Formaldehyde prevented production of methane . Initial methanogenic rate was lower and methanogenesis became exponential when samples were permitted to cool before laboratory incubation . Methanogenesis occurred and methanogenic bacteria were present over a range of 68 to 30 degrees C, with optimum methanogenesis near 45 degrees C . The temperature distribution of methanogenesis in the mat is discussed relative to published results on standing crop, primary production, and decomposition in the thermal gradient . The depth distribution of methanogenesis was similar to that of freshwater sediments, with a zone of intense methanogenesis near the mat surface . Methanogenesis in deeper mat layers was very low or undetectable despite large numbers of viable methanogenic bacteria and could not be stimulated by addition of anoxic source water, sulfide, or a macronutrient solution.

Arch Microbiol, 1978 May 30, 117(2), 189 - 96
A comparative analysis of extreme thermophilic bacteria belonging to the genus Thermus; Degryse E et al.; Several extreme thermophilic Gram negative bacteria found in a thermally polluted river in Belgium have been compared with Thermus strains isolated from widely distant geographical areas . This analysis has become possible after the design of a new culture medium (162) . All strains examined (including the isolate successively denominated Flavobacterium thermophilum and Thermus thermophilus) were found to be morphologically identical with strain YT-1 of Thermus aquaticus . The cells are immotile, rod-like, strictly aerobic, catalase and oxidase positive . They produce amylase, hydrolyze gelatin and are confirmed to be highly sensitive towards penicillin . The nutritional pattern of all strains has been analysed extensively, by testing a broad spectrum of possible substrates . The strains display a uniform response to the microbiological tests applied and most probably belong to the same species: Thermus aquaticus.

Mikrobiologiia, 1978 May-Jun, 47(3), 561 - 2
{Thermophilic bacteria, Thermus ruber, that produce a bright orange pigment}; Loginova LG et al.; A non-sporeforming gram-negative bacterium isolated from soils of Africa was shown to belong to the species Thermus ruber in its morphologo-cytological and physiologobiochemical properties . It differed from the Thermus ruber species described elsewhere (Loginova et al., 1975; Loginova and Egorova, 1975) only on colour: the former contained a bright-red pigment while the latter produced bright-orange pigments.

Chest, 1978 May, 73(5), 608 - 12
The absence of a relationship between serum precipitins and pulmonary disease in a community; Dodge RR et al.; To study the role of serum precipitins in respiratory illness in a community, sera obtained from 3,047 residents of Tucson, Ariz., were tested for the presence of precipitating antibody to a battery of antigens . Positive reactions were obtained in 54 subjects (1.8 percent), a lower incidence than has been reported previously . The majority of these subjects were older than 54 years of age, an age distribution significantly older than the entire sample (P less than 0.01) . Pulmonary function among the subjects with positive precipitin reactions was not significantly different from that of the asymptomatic nonsmokers of the entire sample . None of the subjects who were lifelong residents of Arizona had serum precipitins to any of the thermophilic actinomycetes antigens which were used in the testing . These antigens have been found in association with extrinsic allergic alveolitis, most frequently among patients living in the north central United States and were derived from strains of Micropolyspora faeni, Thermoactinomycetes candidus and vulgaris . Each subject with precipitins to one or more of the tested antigens was matched by age, sex, and socioeconomic class with two subjects from the sample who had negative precipitin reactions . The groups did not differ in their prevalence of respiratory symptoms or abnormalities of pulmonary function . We conclude that the presence of precipitating serum antibodies among subjects in a community is not indicative of the presence of immunologic pulmonary disease but merely reflects previous exposure to the tested antigen . In addition, individuals whose sera contain precipitating antibody appear to have no increased tendency to develop other types of pulmonary disease.

J Clin Pathol, 1978 May, 31(5), 423 - 5
Quantitative recovery of spores from thermophilic spore papers; Everall PH et al.; A homogeniser (Stomacher 400) which does not damage bacteria has been used to produce an homogenate of spore papers, from which it was possible to make quantitative assessments of viable spores recoverable from such papers before and after exposure to sterilising procedures.

J Biochem (Tokyo), 1978 May, 83(5), 1231 - 8
Properties of membrane adenosine triphosphatase of the obligately anaerobic bacterium Veillonella alcalescens; Yoshimura F; Some properties of membrane ATPase activity in Veillonella alcalescens were examined . Mg2+ is required for the activity of the enzyme, and Ca2+ also activates the enzyme to some degree . Of the nucleotide triphosphates, GTP and ITP were hydrolyzed to a lesser extent than ATP . The apparent Km for ATP hydrolysis was 0.25 to 0.63 mM . ADP inhibited the enzyme and the kinetic data of its inhibition showed that the presence of ADP resulted in positive cooperativity . The enzyme activity was strongly inhibited by DCCD, azide, fusidic acid and the antibody to purified soluble ATPase from the thermophilic bacterium PS3 . Oligomycin, dinitrophenol, and ouabain showed no significant effect.

Biotechnol Bioeng, 1978 May, 20(5), 665 - 76
Immobilization of Streptomyces flavochromogenes pullulanase on tannic acid and TEAE--cellulose; Ohba R et al.; Pullulanase was immobilized successfully by simple, inexpensive methods that may be useful for industrial application of this enzyme . A tannin--pullulanase(TP) complex was obtained by addition of tannic acid to the culture filtrate of thermophilic Streptomyces flavochromogenes . TP could be bound to TEAE--cellulose (TTCP) . Immobilization in this manner took place with quantitative retention of activity . The immobilized enzymes were stable for more than six months . The optimum temperatures of the native enzyme and TP were both 50 degrees C; that of TTCP was 45 degrees C . In the presence of 5mM Ca2+, the activity of TTCP was increased approximately twofold and the optimum temperature was raised to 50--60 degrees C . Pullulanase was not significantly eluted from TP or TTCP by NaCl solution (0.1--0.5M).

Mol Gen Genet, 1978 Apr 17, 160(3), 247 - 57
Identification of Escherichia coli and Bacillus stearothermophilus ribosomal protein binding sites on Escherichia coli 5S RNA; Zimmermann J et al.; E coli {32P}-labelled 5S RNA was complexed with E . coli and B . stearothermophilus 50S ribosomal proteins . Limited T1 Rnase digestion of each complex yielded three major fragments which were analysed for their sequences and rebinding of proteins . The primary binding sites for the E . coli binding proteins were determined to be sequences 18 to 57 for E-L5, 58 to 100 for E-L18 and 101 to 116 for E-L25 . Rebinding experiments of purified E . coli proteins to the 5S RNA fragments led to the conclusion that E-L5 and E-L25 have secondary binding sites in the section 58 to 100, the primary binding site for E-L18 . Since B . stearothermophilus proteins B-L5 and BL22 were found to interact with sequences 18 to 57 to 100 it was established that the thermophile proteins recognize and interact with RNA sequences similar to those of E . coli . Comparison of the E . coli 5S RNA sequence with those of other prokaryotic 5S RNAs reveals that the ribosomal proteins interact with the most conserved sections of the RNA.

J Biochem (Tokyo), 1978 Apr, 83(4), 1165 - 71
Purification and properties of superoxide dismutase from Thermus thermophilus HB8; Sato S et al.; Manganese-containing superoxide dismutase was isolated from an extreme thermophile, Thermus thermophilus HB8 . About 150 mg of the enzyme was obtained from 500 g of wet cells . The enzyme was easily crystallized in octahedra from ammonium sulfate solution . The molecular weight of the enzyme was determined to be 8.2 X 10(4) and 8.4 X 10(4) by sedimentation equilibrium and gel-filtration, respectively . The enzyme contains 2 atoms of manganese per mole and consists of four subunits of identical molecular weight, about 2.1 X 10(4) . The amino acid composition of the enzyme is similar to that of the superoxide dismutase of Thermus aquaticus . Proline was detected as the N-terminal amino acid . The isoelectric point was determined to be pH 6.0 by the electrofocusing method . The enzyme has maxima at 283 nm and 480 nm in the absorption spectrum . The CD spectrum suggests that the enzyme has a high alpha-helical content.

Biosystems, 1978 Apr, 10(1-2), 19 - 28
Phylogenetic affinities between eukaryotic cells and a thermophilic mycoplasma; Searcy DG et al.; Thermoplasma acidophilum, a thermophilic mycoplasma, has several unusual features suggesting a possible relationship to eukaryotic cells . One feature is a histone-like protein that is associated with the DNA, condensing it into subunits similar to those in eukaryotic chromatin . A second feature is an association of cytoplasmic proteins that resembles eukaryotic actin and myosin . These two components are widely distributed in different groups of eukaryotic cells, but are typically lacking in prokaryotic cells . Furthermore, T . acidophilum lacks cytochromes and respires by enzymes that apparently are not coupled to oxidative phosphorylation . This primitive type of respiration resembles that of microbodies, another feature which is represented in the cytoplasm of all groups of eukaryotic cells . Furthermore, since T . acidophilum lacks a cell wall and appears to have a primitive correlate of endocytosis, it would appear to be mechanically capable of acquiring a symbiotic mitochondrion . Thus, our observations are consistent with the symbiotic hypothesis for the origin of eukaryotic cells . We suggest that an organism similar to T . acidophilum was the host cell for the original symbiosis, becoming the nucleus and cytoplasm of modern eukaryotic cells.

Mikrobiologiia, 1978 Mar-Apr, 47(2), 371 - 3
{Removal of the RNA from Pseudomonas thermophila cells}; Emnova EE; The RNA content in the cells of Pseudomonas thermophila K-2 grown at 50 degrees comprise 13% of dry weight during exponential growth and 5% of dry biomass in the stationary phase . 60% RNA is degraded after 10 min exposure to heat shock at 70 degrees and subsequent incubation at 50 degrees for 2 hours.

Mikrobiologiia, 1978 Mar-Apr, 47(2), 286 - 92
{Variability in the amylolytic thermophilic bacterium, Bacillus diastaticus, induced by ultraviolet rays}; Murgina VP; The UV-induced variability of the amylolytic thermophilic Bacillus diastaticus 13 was studied . The biosynthesis of amylase was found to very under the action of UV from 2.2 to 158.7% A "plus" variant referred to as the mutant UV 1 was produced at a dose of UV equal to 41.8-10(2) erg/mm2 . Its further selection without using a mutagen made it possible to select a variant UV 1-25 which surpassed the parent strain by 43.3% in amylase biosynthesis . UV-irradiation produced also two mutants with a low activity in amylase biosynthesis . The requirement of growth factors was studied with several mutants.

Mikrobiologiia, 1978 Mar-Apr, 47(2), 250 - 2
{Nucleotide makeup of the DNA of thermophilic bacteria of the genus Thermus}; Aleksandrushkina NI et al.; The nucleotide composition of DNA was determined in extreme thermophilic and obligate thermophilic nonsporeforming bacteria belonging to a new genus Thermus . The GC content (in mol%) in the DNA of exteme thermophilic bacteria varied from 65.3 to 70.8 per cent depending on the strain . The amount of GC (in mol% in the DNA of obligate thermophilic and extreme thermophilic bacteria of the Thermus genus was higher than that in the DNA of sporeforming obligate thermophilic Bac . coagulans . Bac . circulans and Bac . stearothermophilus.

Am J Clin Nutr, 1978 Mar, 31(3), 532 - 40
Assay for free and total choline activity in biological fluids and tissues of rats and man with Torulopsis pintolopessi; Baker H et al.; The sensitive, specific growth response to choline activity of the thermophilic enteric yeast Torulopsis pintolopessi enables estimation of free and bound choline activity in rat and human fluids and tissues- as little as 10 ng/ml of choline is measurable . Unlike other microbial assays, estimation of unbound (free) choline activity is not influenced by methionine or phospholipids . The method also distinguishes differences in choline activity of fluids and tissues from choline-deficient and choline-replete rats . Free and bound choline activity in blood, red blood cells, plasma, and liver from choline-deficient rats were almost 2-fold lower than from choline-supplemented animals . Free and bound choline activity in whole brain from choline-deficient rats were signifigantly higher (more than 2-fold) . The application of the T . pintolopessi method in studying choline status in man and reasons for high choline activity in brain of choline-deficient rats are suggested.

Biochim Biophys Acta, 1978 Feb 15, 532(2), 337 - 46
Alkaline isomerization of thermoresistant cytochrome c-552 and horse heart cytochrome c studied by absorption and resonance Raman spectroscopy; Kihara H et al.; The structure of the thermoresistant cytochrome c (552, Thermus thermophilus) has been investigated at neutral and alkaline pH by absorption and resonance Raman spectroscopy and compared with that of horse heart cytochrome c . The ligands of the ferricytochrome c-552 at neutral pH are considered to be histidine and methionine, whereas the ligands of ferrocytochrome c-552 are histidine and another nitrogen base, histidine or lysine . Ferric cytochrome c-552 undergoes an alkaline isomerization with a pK of 12.3 (25 degrees C), accompanied by a ligand exchange . Horse heart cytochrome c has at least three isomerization states at alkaline pH (pK 9.3, 12.9 and greater than 13.5 at 25 degrees C) . The replacement of the sixth ligand may not be involved in the second isomerization . The thermodynamic parameters for the isomerization were also estimated . The entropy change upon isomerization of cytochrome c-552 is negative, whereas for that of horse heart cytochrome c the entropy change is positive.

J Biol Chem, 1978 Feb 10, 253(3), 850 - 5
Purification and properties of the endonuclease specific for apurinic sites of Bacillus stearothermophilus; Bibor V et al.; An endonuclease specific for apurinic sites when double-stranded DNA is used as substrate has been isolated from the thermophilic bacterium, Bacillus stearothermophilus; it is a monomeric protein of about 28,000 daltons, without action on normal DNA strands or on alkylated sites . The enzyme is quite thermoresistant in the presence of other proteins, has an optimal temperature of 60 degrees, needs monovalent cations for optimal activity, is insensitive to EDTA, and is inhibited by divalent cations; it has no associated exonuclease activity . These latter properties are closer to those of Escherichia coli thermoresistant endonuclease IV, which is also insensitive to EDTA and has no exonuclease activity, and very different from those of the main endonuclease for apurinic sites of the same bacterium . The B . stearothermophilus enzyme is more resistant to urea and detergents than the main E . coli endonuclease for apurinic sites and has a higher content of hydrophobic amino acids.

J Gen Microbiol, 1978 Feb, 104(2), 193 - 9
Isolation of extrachromosomal deoxyribonucleic acids from extremely thermophilic bacteria; Hishinuma F et al.; Eight strains of thermophilic bacteria were examined for the presence of covalently closed circular deoxyribonucleic acid molecules by caesium chloride-ethidium bromide density gradient centrifugation . Four of the eight strains tested, Thermus flavus BS1, AT61, AT62 and Thermus thermophilus HB8 carried covalently closed circular DNA molecules . Thermus flavus BS1 haboured two species of plasmids with molecular weights of 6.1 X 10(6) and 17.0 X 10(6) as determined by electron microscopy . Thermus thermophilus HB8, T . flavus AT61 and T . flavus AT62 carried plasmids with molecular weights of 6.2 X 10(6), 6.6 X 10(6) and 6.6 X 10(6), respectively . Plasmids from T . flavus AT61 and AT62 were indistinguishable in their electrophoretic patterns in agarose or acrylamide gel after digestion with various restriction endonucleases . This is the first evidence for the presence of plasmids in extremely thermophilic bacteria, though their functions are unknown.

Nucleic Acids Res, 1978 Feb, 5(2), 591 - 8
Enzymatic synthesis of a segment of bacteriophage Qbeta coat protein gene; Kikuchi Y et al.; The oligoribonucleotide, A-A-A-C-U-U-U-Gp, constituting a segment of RNA bacteriophage Qbeta coat protein gene was efficiently synthesized at a milligram scale by a combination of enzymatic methods using bacteriophage T4 RNA ligase and the thermophilic polynucleotide phosphorylase . A-A-A-Cp was synthesized from A-A-A and pCp by the newly developed mononucleotide addition method using T4 RNA ligase in a yield of 83%, followed by dephosphorylation with bacterial alkaline phosphatase to obtain A-A-A-C . pU-U-U-Gp was synthesized from pU-U-U and GDP by the simultaneous action of polynucleotide phosphorylase and RNase T1 in a yield of 32% . finally, the two oligonucleotides (A-A-A-C and pU-U-U-Gp) were ligated with T4 RNA ligase and the octanucleotide, A-A-A-C-U-U-U-Gp, was obtained in a yield of 85%.

J Biochem (Tokyo), 1978 Feb, 83(2), 629 - 31
C-type cytochromes isloated from an extreme thermophile, Thermus thermophilus HB8; Hon-Nami K et al.; Two cytochromes of the C-type, c-554 and c-549, were isolated from the soluble fraction of an extreme thermophile, Thermus thermophilus HB8 . Highly purified cytochrome c-554 had absorption maxima at 554, 522, and 417 nm in the reduced state, and at 410 nm in the oxidized state . The alpha-band of the reduced state resembled that of "split-alpha" cytochromes . The isoelectric point was at pH 4.9, and the molecular weight was about 29,000 . Cytochrome c-549, partially purified, had absorption maxima a6 549,520, and 416 nm in the reduced form, and at 408 nm in the oxidized form . The molecular weight was about 25,000 . Both were slowly auto-oxidizable, and did not combine with CO.

Cell, 1978 Feb, 13(2), 329 - 34
Mistranslation in a eucaryotic organism; Palmer E et al.; Previous work from our laboratory has demonstrated that a subclass of the aminoglycoside antibiotics, those containing the drug fragment paromamine, stimulates mistranslation in cell-free protein-synthesizing systems derived from eucaryotic cells . We report here experiments which show that the ciliate Tetrahymena thermophila (formerly T . pyriformis, syngen 1) is sensitive to the paromamine-containing aminoglycoside antibiotics . The drugs are active with respect to growth inhibition, inhibition of protein synthesis in the whole organism, inhibition of protein synthesis in vitro and the stimulation of mistranslation in cell-free protein-synthesizing systems . Because of their misreading properties, these drugs may be useful in isolating and propagating strains carrying mutations which can be translationally suppressed (that is, missense and nonsense mutations).

Appl Environ Microbiol, 1978 Feb, 35(2), 258 - 63
Purification and some properties of riboflavin synthetase from Bacillus stearothermophilus ATCC 8005; Suzuki Y et al.; A riboflavin synthetase was purified 51-fold from a thermophilic organism, Bacillus stearothermophilus ATCC 8005, that grew at 40 to 72 degrees C . Some of the properties of the enzyme are: (i) its temperature optimum was 95 degrees C, and the activity was negligible below 40 degrees C; (ii) the Arrhenius plot of the initial reaction rates was concave upward, with a break at 65 degrees C, and the apparent activation energies below and above 65 degrees C were 4.2 X 10(4) and 6.7 X 10(4) J/mol, respectively; (iii) the enzyme was fairly stable up to 60 degrees C without 6,7-dimethyl-8-ribityllumazine; this substance protected the enzyme from inactivation above 60 to 97 degrees C; (iv) the pH range for stability was 6.0 to 10.0 at 26 degrees C and 6.3 to 7.6 at 55 degrees C; (v) the enzyme was highly resistant at 26 degrees C to denaturation in 8 M urea, but the tolerance was extremely low at 55 degrees C; (vi) its molecular weight was estimated at 45,000; (vii) the Km for 6,7-dimethyl-8-ribityllumazine was 23 micrometer at 55 degrees C and 29 micrometer at 75 degrees C; (viii) its pH optimum was 6.7 to 7.2; (ix) 6-methyl-7-hydroxy-8-ribityllumazine was a competitive inhibitor (Ki = 0.18 micrometer); (x) the activity was sensitive to heavy-metal ions and thiol reagents; (xi) the enzyme did not require cofactor or a carbon donor; and (xii) the molar ratio of 6,7-dimethyl-8-ribityllumazine consumption to riboflavin formation was 2 throughout the entire reaction . Properties i through vi distinguish this enzyme from riboflavin synthetases purified by other investigators from mesophilic organisms, Ashbya gossypii, Eremothecium ashbyii, Escherichia coli, yeast, and spinach.

Microbios, 1978, 23(93-94), 193 - 8
Improved growth of Thermus aquaticus on cellular lysates; Stramer SL et al.; Studies on the thermophilic bacterium Thermus aquaticus have demonstrated the ability of this organism to exist on its own cell lysates . The use of 0.1% tryptone, and 0.1% yeast extract and a mineral salts lysate medium increased growth 9-to 44-fold over the above medium without cell lysates . It has also been established in this study that, due to the high growth temperature of T . aquaticus, its metabolic activities are greatly accelerated resulting in early cell destruction . Therefore, bacterial lysates from which other cells could derive nutrients and/or growth factors would constantly be available in the environment immediately around the organism to aid the growth of cells in natural as well as man-made thermal environments.

Poumon Coeur, 1978, 34(3), 209 - 17
{Two surveys on farmer's lung disease and bird breeder's disease in the Pyrenees region}; Seguela JP et al.; Two different surveys, one strictly epidemiological in rural areas, the other retrospective and within hospitals, led to a better knowledge of the setting of the FLD and BBD in South Pyrenees region . In the country, more than 12% of patients had specific precipitins, 6% had either one or both diseases; women had anti-aviary precipitins more frequently than men . The retrospective hospital survey yielded 456 cases of patients admitted or consulting revealing numerous correlations between the different clinical, radiological and functional syndromes on the one hand and the serological positivity on the other . Serological diagnosis was most useful, as a wide variety of antigenic extracts was used (actinomycetes thermophiles, mouldy hay, bird droppings), mixed sensitizing was extremely frequent . The widely found positivity towards avian antigens seems specific to the South Pyrenees region.

Mikrobiologiia, 1978 Jan-Feb, 47(1), 142 - 8
{Thermophillic sulfate-reducing bacteria from oil-bearing strata}; Nazina TN et al.; The paper describes pure cultures (strains 435 and 781) of thermophilic spore-forming sulphate-reducing bacteria isolated from oil strata . The strain 435 was classed as a new subspecies Desulfotomaculum nigrificnas subsp . salinus according to its morphological, physiological, and biochemical characteristics . The cells of the culture are rod-like, 2-5 mcm long and 0.9-1.3 mcm thick . Some cells are spindle-shaped . The cells have peritrichously arranged flagella . Spores are oval, located terminally or subterminally, slightly widening the cells . The culture is an obligate anaerobe . It grows in media containing sulphates and assimilates sodium salts of lactic, pyruvic, malic and formic acids, ethanol, and butanol . The cuture assimilates pyruvate in the absence of sulphates . Sulphate, sulphite and thiosulphate are electron acceptors when the culture grows on lactate . The organism can grow at 40-70 degrees C, the optimum temperature being 60 degrees C . It requires NaCl for growth, and can grow even in the presence of 4% NaCl in the medium . The optimum should contain 1% NaCl and microelements . Desulfoviridin is absent . The cells contain a cytochrome of the protoheme class . The content of G + C base pairs in DNA is 57.0 +/- 0.5 mol . %.

Mikrobiologiia, 1978 Jan-Feb, 47(1), 101 - 6
{Natural variability of Bacillus diastaticus, a thermophilic bacterium that forms amylase}; Murygina VP; Natural variability of the thermophilic culture of Bacillus diastaticus was studied, and the population of its cells was found to be genetically heterogeneous . The bulk (75.5%) of the cells had a high activity of amylase biosynthesis; 21.5% of the cells had a low activity; 2.94% of the cells had no activity . One inactive variant and two variants whose activity was higher that that of the parent culture with respect to amylase biosynthesis have been selected.

Zentralbl Bakteriol Naturwiss, 1978, 133(5), 385 - 93
{Studies on the composting of feathers (author's transl)}; Kuster E et al.; The nitrogen of keratin of feathers is not taken up by plants as a suitable N-source . It becomes available by the keratinolytic activity of microorganisms . The high temperature evolved during the composting process promotes the growth and activity of thermophilic, keratin-decomposing microorganisms, e.g . Thermoactinomyces vulgaris . The increasing in temperature (self-heating) reflects the course of composting . A feather-peat mixture exhibits a considerably high heat production, i.e . a fast decomposition of keratin and other organic matter.

C R Seances Soc Biol Fil, 1978, 172(2), 314 - 9
Induced resistance to 6-methylpurine and cycloheximide in Tetrahymena . II . Variation within vegetative cultures of micronucleate T . thermophila and amicronucleate T . pyriformis; Byrne BC; Resistance to 6-methylpurine and cycloheximide has been induced in both the micronucleate species Tetrahymena thermophila and the amincronucleate species T . pyriformis . Resistance follows only after mutagen treatment and vegetative growth . The frequencies with which resistant variants are induced and the independence of mutagenesis and selection are demonstrated . All evidence is consistent with the hypothesis that macronuclear subunits are assorting in both species during vegetative growth to produce new phenotypes among subclones.

Zentralbl Bakteriol Naturwiss, 1978, 133(7-8), 713 - 22
Identity and lipase activity of an isolate of Thermoactinomyces vulgaris; Elwan SH et al.; An actinomycete with thermophilic nature and strong lipolytic activity was isolated by enrichment from an Egyptian soil . It proved to be Thermoactinomyces vulgaris . Variabilities justifying Adansonian systematics were reported and discussed . The potential relation of an occasional drop in successfully resumed activity to an auto-inhibition phenomenon has been notified . Optimum conditions for the activity of the extracellular lipase(s), including stability, suggest the practical importance of the enzyme(s) in the growth filtrates.

Adv Biophys, 1978, 10, 209 - 47
Proton translocating ATPase: its pump, gate, and channel; Kagawa Y; Proton translocating ATPase of oxidative phosphorylation was divided into three functional units: pump, channel, and gate . This was achieved by the use of highly stable pure ATPase obtained from a thermophilic bacterium PS3 . The pump and gate were found in a catalytic moiety of the ATPase called TF1, and the channel was in the remaining hydrophobic moiety of the ATPase called TF0 which rendered TF1 sensitive to energy transfer inhibitor such as DCCD . TF1 was composed of five subunits (alpha, 56,000; beta, 53,000; gamma, 32,000; delta, 15,500; epsilon, 11,000 daltons) . The essential component of the pump was beta-subunit, since beta gamma-complex or alpha beta delta-complex showed ATPase activity . The gate which blocked passive leakage of protons through TF0 in the proteoliposomes was shown to be gamma delta epsilon-complex in TF1 . Both delta- and epsilon-subunits were required to connect alpha beta gamma-complex to TF0 . TF0 was identical to the channel and was composed of three kinds of subunits (19,000, 13,500, and 5,400 daltons) and the smallest one was {14C}-DCCD binding protein . When the ATPase was incorporated into vesicles containing highly stable saturated branched phospholipids, ATP-driven electrochemical potential of proton (delta mu H+ = 253mV) and proton gradient driven net synthesis of ATP were demonstrated . For these activities, pump, channel, and gate of proton translocating ATPase were all required.

JAMA, 1977 Dec 19, 238(25), 2705 - 8
Allergic alveolitis caused by home humidifiers . Unusual clinical features and electron microscopic findings; Burke GW et al.; Allergic alveolitis (hypersensitivity pneumonitis) developed in two patients who were exposed to home humidifiers contaminated by thermophilic actinomycetes . Diagnosis was difficult because severe dyspnea was chronic and not easily associated with specific environmental exposure . Furthermore, chest roentgenograms were normal for long periods despite severe physiologic abnormalities . After several years of disabling symptoms, open lung biopsy findings suggested allergic alveolitis rather than sarcoidosis because a mononuclear interstitial infiltrate overshadowed the granulomas . Also, plasma cells were prominent and there was an intense bronchiolitis . Cultures of the humidifier water grew thermophilic organisms to which the patient's sera formed precipitins . Both patients experienced notable subjective and objective improvement following removal of the contaminated appliances . The ultrastructure of alveolar macrophages and basement membranes was similar to that described in animal models, suggesting cell-mediated hypersensitivity in the patients.

Biochim Biophys Acta, 1977 Dec 8, 485(2), 417 - 23
Thermostability at ultrahigh temperatures of thermolysin and a protease from a psychrotrophic Pseudomonas; Barach JT et al.; Thermal inactivation at 110-150 degrees C of thermolysin (EC 3.4.24.4), produced by the thermophile Bacillus thermoproteolyticus, and the extracellular protease of Pseudomonas sp . MC60 a psychotroph, were investigated at 130 degrees C, both enzymes had approximately the same deltaH (22 kcal/mol) and deltaS (-13.5 cal/mol per degree) values . Both enzymes contain zinc and calcium . The amino acid compositions of the enzymes were similar except that MC60 protease exhibited a more typical tyrosine content . Comparable heat resistance at extreme temperatures of enzyme produced by psychrotrophic and thermophilic organisms emphasizes the difference between molecular properties that resist denaturation at elevated temperatures and those that allow reversible denaturation.

Appl Environ Microbiol, 1977 Dec, 34(6), 715 - 9
Production of thermophilic actinomycete-hay aerosols for use in experimental hypersensitivity pneumonitis; Burrell R et al.; An investigation of the factors that influence the production of dense aerosols of Micropolyspora faeni and Thermoactinomyces vulgaris from hay cultures revealed that the density, freshness, and moisture content of the hay were important influences . Dry aerosols were produced under optimum conditions from hay cultures of either actinomycete and from sterile hay for inhalation challenges to unimmunized rabbits . Depressions in arterial oxygen tensions and hemolytic complement were monitored after such challenge . This investigation showed that hay cultures are capable of inciting a hypersensitivity-like reaction in the lungs, even in the absence of immunization to the organisms contained within the hay, but the entire hypersensitivity reaction cannot be attributed to M . faeni and T . vulgaris alone . Hay itself or other microbial or chemical components appear to have some heat-stable component that possibly contributes to the pathogenesis of the disease.

Appl Environ Microbiol, 1977 Dec, 34(6), 626 - 9
Effect of growth substrate on thermal death of thermophilic bacteria; Merkel GJ et al.; The heat sensitivity of gram-negative, hydrocarbon-utilizing thermophilic bacteria was altered by a change in growth substrate . Thermophilic strains CC-6, BI-1, and LEH-1, grown with acetate or n-heptadecane as the carbon source, had a higher survival rate when incubated 5 degrees C above their maximum growth temperature than cells of the same organism after growth on glucose or glycerol . There was a correlation between the growth substrated, heat resistance, and the ratios of cellular n-hexadecanoic acid/branched hexadecanoic acid and n-heptadecanoic acid/branched heptadecanoic acid . The bacterial cells that were more heat resistant had ratios of straight-chain/branched-chain fatty acids above 1.0, whereas the heat-sensitive cells had ratios below 0.6.

J Biochem (Tokyo), 1977 Dec, 82(6), 1751 - 8
Formations of electrochemical proton gradient and adenosine triphosphate in proteoliposomes containing purified adenosine triphosphatase and bacteriorhodopsin; Sone N et al.; Proteoliposome vesicles containing both bacteriorhodopsin of Halobacterium halobium and H+-translocating ATPase {EC 3.6,1.3} of a thermophilic bacterium, PS3, (TF0-F1) were reconstituted by either the dialysis method or the sonication method . Generation of the electrochemical proton gradient (deltamuH+) in these vesicles was measured using 9-aminoacridine for estimation of the chemical (deltapH) component and 8-anilinonaphthalene sulfonate for the electrical (deltaphi) component) . In illuminated bacteriorhodopsin-vesicles the deltamuH+ reached 180-190 mV when reconstituted by the dialysis method and 210-220 mV when reconstituted by the sonication method . Vesicles reconstituted from both TF0-F1 and bacteriorhodopsin by the dialysis method generated a deltapH+ of about 200 mV on addition of ATP, while vesicles prepared by the sonication method generated very little deltamuH+, if any . These vesicles generated similar deltamuH+ on illumination to that found in bacteriorhodopsin-vesicles . Using vesicles reconstituted from both TF0-F1 and bacteriorhodopsin by the dialysis method, light dependent ATP synthesis was measured in relation to deltamuH+ formation . It was necessary to generate a deltamuH+ of above 170 mV for demonstration of appreciable formation of ATP and the greater the deltamuH+, the faster the rate of ATP synthesis.

MMW Munch Med Wochenschr, 1977 Nov 11, 119(45), 1463 - 6
{"No-touch" measurement of skin temperature in cerebral ischemia and lesions of the sympathetic trunk . Clinical and technical testing of a new infrared apparatus for measuring skin temperature (Ultrakust Thermophil M 202) (author's transl)}; Haller P et al.; The clinical test was carried out in extracranial carotid occlusions and acute peripheral paralysis with sympathetic lesions . The measurements obtained were compared with those of the infrared thermovision . Both in the technical and in the clinical tests, the device proved to be very suitable . The practical procedure of temperature measurement (cerebral ischemia and lesions of the sympathetic trunk) is described.

J Biochem (Tokyo), 1977 Nov, 82(5), 1457 - 61
Bacteriophage phiNS11: a lipid-containing phage of acidophilic thermophilic bacteria . III . Characterization of viral components; Sakaki Y et al.; Components of a lipid-containing phage phiNS11 were characterized . The phage had five protein components, the molecular weights of which were 59,000, 44,000, 33,000, 23,000, and 18,000 . Viral lipid consisted of six components, which were also found in the host bacterial lipid . The relative amounts of these viral lipid components were very similar to those of the bacterial lipid . The phage contained omega-cyclohexyl fatty acids characteristic of Bacillus acidocaldarius as the main fatty acids . The phage nucleic acid was a linear double-stranded DNA, the molecular weight of which was 9.3--9.4 X 10(6) daltons . The guanine plus cytosine content of the DNA was determined to be about 52% from chemical analysis, buoyant density (1.711 g/cm3 in CsCl) and melting temperature (90.6 degrees C in 0.15 M NaCl plus 0.015 M sodium citrate) . The phage contained two kinds of polyamine; spermidine and spermine.

J Biochem (Tokyo), 1977 Nov, 82(5), 1451 - 6
Bacteriophage phiNS11: a lipid-containing phage of acidophilic thermophilic bacteria . II . Purification and some properties of the phage; Sakaki Y et al.; An improved procedure for the purification of a lipid-containing acidophilic thermophilic bacteriophage, phiNS11, is described . The procedure involves Cs2SO4 discontinuous density gradient, Cs2SO4 equilibrium density gradient and sucrose gradient centrifugations . The phage has an icosahedral shape, 75 nm in diameter, with spike-like structures radiating from the vertices . It contained about 12% by weight of lipid, 13--14% of DNA and 74--75% of protein . The phage was stable between pH 2 and 5, but rapidly inactivated above pH 6 . It was highly sensitive to organic solvents and detergents . Some other basic properties such as the buoyant density and UV spectrum of the purified phage are also described.

Differentiation, 1977 Oct 20, 9(1-2), 119 - 30
Perturbance analysis of nuclear determination in Tetrahymena I: background, rationale, and illustrative example, employing temperature responses; Nanney DL et al.; A model based on comparative considerations, is presented for the nuclear determination of mating type in Tetrahymena thermophila . The model proposes a system of three binary control elements, each capable of stable persistence in one of two states . A general method is proposed for evaluating the model and for assigning particular mating types to particular compound states . Preliminary assignments of mating types are made from the responses of nuclei to temperature differentials.

Mycopathologia, 1977 Sep 16, 61(2), 85 - 91
{Different behavior of 22 strains of Emmonsia Cifferi and Montemartini 1959, a monilial fungus, in the lungs of laboratory mice in comparison with their parasitic morphology in vitro}; Boisseau-Lebreuil MT; The 22 strains of Emmonsia Ciferri & Montemartini 1959, inoculated intranasally to laboratory mice are not equally virulent . One month after the inoculation, 15 of the strains had produced adiaspores 120 t9 190 micrometer in diameter in the lung . Another strain produced adiaspores measuring 44 micrometer and 2 others measuring 20 micrometer or 10 micrometer . The remaining 4 strains did not develop in the lung tissue . Four thermophilic strains, which in vitro have adiaspores measuring 8 to 15 micrometer, had adiaspores reaching 120-180 micrometer in vivo . Neither budding nor endosporulation could be observed in any adiaspore.

J Biol Chem, 1977 Sep 10, 252(17), 6125 - 31
Purified proton conductor in proton translocating adenosine triphosphatase of a thermophilic bacterium; Okamoto H et al.; 1 . The membrane-integrated portion (TF0) of the proton translocating ATPase complex (TF0-F1) of the thermophilic bacterium PS3 was highly purified . Its proton-conducting activity was investigated in vesicles reconstituted from TF0 and phospholipids (TF0 vesicles) . 2 . The rate of proton conduction through TF0 was proportional to the membrane potential imposed (6H+ uptake/s/TF0 molecule with 103 mV at pH 8.0) . The pH profile of the rate revealed that a proton, not a hydroxy ion, was the true substrate conducted and that there was a monoprotic proton binding site in TF0 (pKa = 6.8) . The temperature coefficient of proton conductance of TF0 showed a considerable variation depending on the phospholipids of the vesicles with respective transition temperatures . 3 . Passive proton conduction through TF0 was inhibited stoichiometrically by addition of either the soluble ATPase portion (TF1) of TF0-F1, or an energy transfer inhibitor dicyclohexylcarbodiimide or an antibody against TF0 . 4 . The proton conductance of TF0 was concluded to represent its intrinsic activity in the original TF0-F1 complex.

Eur J Biochem, 1977 Sep, 78(2), 599 - 608
Structural fluctuation of the polypeptide-chain elongation factor Tu . A comparison of factors from Escherichia coli and Thermus thermophilus HB8; Ohta S et al.; The kinetics of hydrogen-deuterium exhcange in the polypeptide chain elongation factor Tu (EF Tu) from Escherichia coli and that from Thermus thermophilus HB8 has been examined in aqueous solutions at various pH and temperatures by means of infrared absorption measurements . The free EF-Tu from E . Coli has a greater reaction rate at all pH values and at every temperature than that of the GTP-bound or GDP-bound EF-Tu . The free EF-Tu from T . thermophilus, on the other hand, has an alomst equal reaction rate to that of EF-Tu-GDP in the temperature range 38-55 degrees C . For the peptide NH groups belonging to a medium-labile kinetic class, a small but definite difference in the rate of exchange reaction was observed between EF-Tu-GDP and EF-Tu-GTP for both E . coli and T . thermophilus . For less labile peptide NH groups, on the other hand, the rate of the exchange reaction with EF-Tu-GDP from T . thermophilus is only slightly affected by the pH of the solution at 38 degrees C and 45 degrees C, while the rate constant(k) with E . coli EF-Tu-GDP is pH-dependent (log k oc pH) . For T . thermophilus EF-Tu, heat stability measurements, kinetics of the rates of GDP and GTP dissociation, and circular dichroic measurements have also been made . The molecular basis for the thermostability of T . thermophilus EF-Tu is discussed.

J Allergy Clin Immunol, 1977 Sep, 60(3), 169 - 73
Correlation of exposure to various respiratory pathogens with farmer's lung disease; Marx JJ Jr et al.; Complement-fixing antibodies (CFA) to a panel of microorganisms commonly associated with respiratory disease were measured in a number of agricultural populations . The panel included Mycoplasma pneumoniae, influenza viruses A and B, parainfluenza virus types 1, 2, and 3, adenovirus, and respiratory syncytial virus . The agricultural populations were grouped according to a clinical history of farmer's lung disease (FLD) and the presence of antibodies to the thermophilic actinomycetes (TA) . Farmers with precipitating antibody activity to one or more of the TA (groups I and II) demonstrated a greater frequency of CFA to M . pneumoniae and parainfluenza virus types 1, 2, and 3 than those groups without antibody to the TA, but the level of CFA was not higher in the positive subjects . Immunoglobulin levels were also elevated in groups I and II when compared to the control groups . Unlike IgG and IgM, IgA was elevated only in the farmers who had a clinical history of FLD (group I) but not in farmers without a clinical history . The results suggest that farmers who develop FLD are exposed to a wider variety of pathogens than are other farmers, but do not respond in an accelerated manner.

J Mol Evol, 1977 Aug 5, 9(4), 305 - 11
An ancient divergence among the bacteria; Balch WE et al.; The 16S ribosomal RNAs from two species of methanogenic bacteria, the mesophile Methanobacterium ruminantium and the thermophile Methanobacterium thermoautotrophicum, have been characterized in terms of the oligonucleotides produced by digestion with T1 ribonuclease . These two organisms are found to be sufficiently related that they can be considered members of the same genus or family . However, they bear only slight resemblance to "typical" Procaryotic genera; such as Escherichia, Bacillus and Anacystis . The divergence of the methanogenic bacteria from other bacteria may be the most ancient phylogenetic event yet detected--antedating considerably the divergence of the blue green algal line for example, from the main bacterial line.

Biochem J, 1977 Aug 1, 165(2), 247 - 53
Purification and properties of histidinol dehydrogenases from psychrophilic, mesophilic and thermophilic bacilli; Lindsay JA et al.; As a first step in elucidating one molecular mechanism of adaptation to life at extreme temperatures, we purified and characterized the enzyme histidinol dehydrogenase (EC 1.1.1.23) from a number of bacilli whose growth temperatures range from 5 degrees t to 90 degrees C . The enzymes were purified by (NH4)2SO4 precipitation, ion-exchange chromatography on Sephadex, affinity chromatography on histamine- or histidine-Sepharose and preparative gradient gel electrophoresis . All had similar mol.wts . (29200), sedimentation coefficients (S20,w 2.56S), affinities for histidinol and NAD+ (Km = 48 micron and 0.2 mM respectively) and all had pH optima at 9.6 . Marked differences were observed in stability with respect to temperature and the temperature at which the initial velocity for histidinol dehydrogenation was optimal . These optima range from 25 degrees C for the enzyme from the psychrophilic species through to 41 degrees C for the mesophiles to 85-92 degrees C for the extreme thermophiles . It is concluded that the ability of the enzymes to operate at their various optimum temperatures is an intrinsic property of their amino acid sequences.

Eur J Biochem, 1977 Aug 1, 77(3), 575 - 83
Thermophilic polynucleotide phosphorylase from Thermus thermophilus . Purification and properties of an altered form of enzyme which lacks phosphorolytic activity to polynycleotide; Hishinuma F et al.; A thermophilic polynucleotide phosphorylase lacking polynucleotide phosphoryltic activity was purified from Thermus thermophilus HB-8 strain . The enzyme is an altered form of the native polynucleotide phosphorylase, probably attacked by the proteinase(s) of this extreme thermophile during the purification process . This modified enzyme lacks phosphorolytic activity to poly(A) while retaining weak activity to phosphorolyse tetranucleotides or hexanucleotides . The purified enzyme was shown to be homogenous by electrophoretic analysis in polyacrylamide gel . This enzyme had a molecular weight of 190 000 as calculated both from electrophoresis on polyacrylamide gel and from the Stoke's radius derived from the gel filtration pattern and the sedimentation coefficient . The enzyme was separated into three polypeptide chains by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate; their molecular weights were calculated to be 92000, 73000 and 35000 . The enzyme was thermophilic and thermotolerant, exhibiting its maximal activity at 70 degrees C . The four ribonucleoside diphosphates (ADP, GDP, UDP and CDP) were polymerized to the extent of 7-S size.

Can J Microbiol, 1977 Aug, 23(8), 1088 - 95
The sporulation process in Thermomonospora fusca as revealed by scanning and transmission electron microscopy; Crawford DL et al.; The sporulation process in the thermophilic actinomycete Thermomonospora fusca was observed by scanning and transmission electron microscopy . As shown by scanning electron microscopy, spores were produced primarily on aerial hyphae and first appeared as bud-like enlargements at the tips of short multibranched sporophores . Young spores were oval to spherical in shape with a smooth surface . As they matured spores enlarged and developed a rough and globular covering, which was quite fragile and easily detached from the spore . This outer layer, as observed by transmission electron microscopy, was thought equivalent to the sheath of other Thermomonospora species . In cross section, mature spores exhibited a thick spore coat underneath the outer globular layer . This spore coat was usually observed as a single layer, but some spores produced a bilayered coat . No multilayered spore coat or spore cortex was observed in the heat-sensitive spores of T . fusca . They were, therefore, shown to be aleuriospores (microcondia), and not endospores.

J Biochem (Tokyo), 1977 Aug, 82(2), 437 - 47
Purification and properties of the polypeptide chain release factor (RF-4) from an extreme thermophile, Thermus thermophilus HB8; Ota Y et al.; The polypeptide chain release factor 1 (RF-1) has been purified from an extreme thermophile, Thermus thermophilus HB8 . The purification procedure included steps of aqueous two-phase partition, ammonium sulfate fractionation, and column chromatographies on DEAE-Sephadex, Sephadex G-150, and CM-Sephadex . The preparation was more than 90% pure as judged by polyacrylamide gel electrophoresis . The specific activity was about 3.3 pmol of formyl-{3H}-methionine released in 1 min at 25 degrees C per microgram of protein under the standard assay conditions using 4 pmol of the initiation complex and 1 nmol of UpApG . The molecular weight as determined by gel filtration on Sephadex G-150 and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, was 58,000 and 45,000, respectively . As expected, the factor was extremely heat-stable, 50% of its activity remaining after incubation for 5 min at 84 degrees C . Several properties of the reaction catalyzed by RF-1 are also described.

Appl Environ Microbiol, 1977 Aug, 34(2), 185 - 8
Production of beta-galactosidase from Streptococcus thermophilus grown in whey; Rao MV et al.; beta-D-Galactosidase (EC 3.2.1.23) was extracted from Streptococcus thermophilus grown in deproteinized cheese whey . Cultural conditions optimum for maximum enzyme production were pH 7.0, 40 degrees C, and 24 h . Proteose peptone (2.0%, wt/vol) and corn steep liquor (2.8%, wt/vol) were highly stimulatory, increasing the enzyme units available in their absence from 660 U/liter of medium to 18,200 and 10,000 U/liter of medium, respectively, in their presence . There was an insignificant increase in the production of enzyme in the presence of added inorganic nitrogen and phosphorus sources . Enzymatic hydrolysis for recuction of lactose content in aqueous solution and in skim milk was studied.

Mikrobiologiia, 1977 Jul-Aug, 46(4), 619 - 24
{Hydrogenase activity of the thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila}; Emnova EE et al.; The hydrogenase activity of the intact cells of a thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila K-2 was determined using methylene blue; it was several times higher than the rate of hydrogen uptake in the presence of oxygene and carbon dioxide . The activity of membrane-associated hydrogenase was assayed with the aid of phenazine methosulphate and 2,6-dichlorphenolindophenol as a cascade electron carrier . The enzyme is sufficiently stable in the air . The stability increases in the atmosphere of hydrogen . The membrane-bound enzyme was activated by Mn2+ ions . The pH-optimum of the enzyme activity in 0.1 M Tris-HCI buffer was 8,5-9,0 . Natural electron acceptors tested, such as NAD, FMN, riboflavin, and cytochrome c, had no effect on the reaction rate . The enzyme is relatively thermostable: its activity was halved after heating at 78 degrees C for 10 min or at 80 degrees C for 8 min . Energy of activation was calculated . It was 14.5 kcal-mol-1 within the range of 23-40 degrees C and 10.3 kcal-mol-1 within the range of 40-60 degrees C.

J Biol Chem, 1977 Jun 25, 252(12), 4256 - 61
Nucleotide sequence of Thermus aquaticus ribosomal 5 S ribonucleic acid . Sequence homologies in thermophilic organisms; Nazar RN et al.; The nucleotide sequence of ribosomal 5 S RNA from Thermus aquaticus grown at 75 degrees is p(A)-A-U-C-C-C-C-G-C-C-C-U-U-A-G-C-G-G-C-G-U-G-G-A-A-C-A-C-C-C-G-U-U-C-C-C-A-U-U-C-C-G-A-A-C-A-C-G-G-A-A-G-U-G-A-A-A-C-G-C-G-C-C-A-G-C-G-C-C-G-A-U-G-G-U-C-A-C-U-G-G-G-A-C-C-G-C-A-G-G-G-U-C-C-U-G-G-A-G-A-G-U-A-G-G-U-G-C-U-G-G-U-G-C-G-G-G-G-A-(U) . The major molecular species is 120 nucleotides long; some molecules are one or two nucleotides shorter with one less nucleotide at either or both termini . When compared to other 5 S rRNAs, the sequence homology was greater with a thermophile bacterium (Bacillus stearothermophilus) than with mesophilic species . The comparisons further indicate that among prokaryotes, eleven of the nucleotide residues in T . aquaticus 5 S RNA may be largely restricted to thermophiles . Possible models for the secondary structure of T . aquaticus 5 S rRNA are discussed with respect to products of limited digestion and the unique nucleotides.

Arch Microbiol, 1977 Jun 20, 113(3), 221 - 9
Temperature range variants of Bacillus megaterium; Stahl S et al.; Facultatively and obligately thermophilic variants were isolated from 3 out of 12 tested mesophilic Bacillus megaterium strains . The variants occurred at a frequency of 10(-8)-10(-9) . The ability to grow at elevated temperatures was cured by means of treatment with acridine orange . Stable revertants were isolated from facultatively and obligately thermophilic variants . An unknown type of megacin was produced by the facultative thermophiles . This megacin attacked mesophilic and obligately thermophilic strains . The thermophiles displayed a few divergent taxonomic characteristics but a close relationship between the strains was indicated by the megacin spectrum and sensitivity to phage . Arrhenius plots revealed that the strains could be considered as temperature range variants and that the temperature characteristic increased with growth at a higher temperature range . The case for a plasmid involvement in the phenomenon is discussed.

Biochim Biophys Acta, 1977 Jun 17, 476(4), 287 - 94
An efficient synthesis of polyguanylic acid by a thermophilic polynucleotide phosphorylase; Kikuchi Y et al.; Polyguanylic acid (poly(G)) was synthesized from GDP in a yield of 60-75% by Thermus thermophilus polynucleotide phosphorylase (polyribonucleotide: orthophosphate nucleotidyltransferase, EC 2.7.7.8) at 70 degrees C, pH 8.5 in the presence of Mg2+ . The yield was dependent on the ratio of GDP to Mg2+, but was independent of the concentrations of enzyme or substrate . The maximal rate of GDP polymerization was obtained when the ratio of GDP to Mg2+ was 3:1 . However, by prolonged incubation, the higher initial ratio of over 4:1 was preferred because of the rapid consumption of GDP in the reaction mixture . Poly(G) prepared by 1 h incubation was heterogeneous in size from 5 S to over 23 S, but by prolonged incubation of 19 h the size of product converged to 9 S as judged by sucrose density gradient centrifugation . Its chain length was determined by terminal nucleoside analysis to be 200 nucleotides long.

Biochim Biophys Acta, 1977 Jun 9, 460(3), 480 - 9
Kinetic studies on redox reactions of hemoproteins . I . Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ferrocyanide; Kihara H et al.; The oxidation-reduction reaction of horse heart cytochrome c and cytochrome c (552, Thermus thermophilus), which is highly thermoresistant, was studied by temperature-jump method . Ferrohexacyanide was used as reductant . (Formula: see text.) Thermodynamic and activation parameters of the reaction obtained for both cytochromes were compared with each other . The results of this showed that (1) the redox potential of cytochrome c-552, + 0.19 V, is markedly less than that of horse heart cytochrome c . (2) deltaHox of cytochrome c-552 is considerably lower than that of horse heart cytochrome c . (3) deltaSox and deltaSred of cytochrome c-552 are more negative than those of horse heart cytochrome c . (4) kred of cytochrome c-552 is much lower than that of horse heart cytochrome c at room temperature.

J Biol Chem, 1977 May 25, 252(10), 3480 - 5
Reconstitution of adenosine triphosphatase of thermophilic bacterium from purified individual subunits; Yoshida M et al.; 1 . Five subunits (alpha, beta, gamma, delta, and epsilon) of an ATPase from a thermophilic bacterium PS3 were purified in the presence of 8 M urea by ion exchange chromatography . Then the ATPase activity was reconstituted by mixing the subunit solutions and incubating them at 20-45 degrees, at pH 6.3 to 7.0 . 2 . Mixtures containing beta + gamma or alpha + beta + delta regained ATP-hydrolyzing activity, but mixtures of alpha + beta and beta + delta did not . Combinations not including beta were all inactive . 3 . The ATPase activity reconstituted from alpha + beta + delta was thermolabile and insensitive to NaN3, whereas the activities obtained from mixtures containing beta and gamma were thermostable and sensitive to NaN3, like the native ATPase . 4 . The assemblies containing both beta and gamma subunits had the same mobility as the native ATPase molecule on gel electrophoresis, those without the gamma subunit moved more rapidly toward the anode . 5 . Subunits epsilon and delta did not inhibit the ATPase activity of either the assembly (alpha + beta + gamma) or the native ATPase.

J Biol Chem, 1977 May 25, 252(10), 3399 - 401
Repressible alkaline phosphatase from Thermus aquaticus: associated phosphodiesterase activity; Smile DH et al.; A repressible alkaline phosphatase has been isolated from the extreme bacterial thermophile . Thermus aquaticus, and has been purified to homogeneity as judged by disc acrylamide electrophoresis and sodium dodecyl sulfate electrophoresis . Upon investigation, the purified enzyme was shown to hydrolyze certain phosphodiesters in addition to a wide variety of phosphomonoesters . The diesters included bis-p-nitro-phenyl phosphate and thymidine 3'-monophospho-p-nitro-phenyl ester . The temperature optimum for the diesterase activity was 80--85 degrees at pH 7.2 . Orthophosphate competitively inhibited both activities . Nucleotides such as AMP, ADP, and ATP also inhibited both esterase activities as did alpha-D-glucose 1-phosphate and alpha-sodium glycerol phosphate . The isoelectric point of the enzyme was determined to be 8.4.

J Biol Chem, 1977 May 10, 252(9), 2956 - 60
Adenosine triphosphate synthesis by electrochemical proton gradient in vesicles reconstituted from purified adenosine triphosphatase and phospholipids of thermophilic bacterium; Sone N et al.; Vesicles were reconstituted from a purified dicyclohexyl-carbodiimide-sensitive ATPase complex (TF0-F1) and phospholipids of a thermophilic bacterium PS3 . These vesicles synthesized ATP from ADP and Pi with energy from an electrochemical proton gradient (delta-micronH+) formed by a pH gradient and an electrical potential across their membranes . Maximal ATP synthesis was achieved by incubating the vesicles in malonate at pH 5.5 with valinomycin, and then rapidly transferring them to a solution of pH 8.4 and 150 mM K+ . Under these conditons ATP synthesis continued at a decreasing rate for 30 s at 40 degrees . Appreciable formation of ATP (40 to 150 nmol/mg of TF0-F1) occurred at an initial delta-micronH+ above 205 mV and moderate formation at an initial value above 180 mV . ATP hydrolysis by the vesicles produced a delta-micronH+, and the additions of 32Pi and hexokinase to them resulted in 32Pi esterification . Analysis of the time courses of 32Pi esterification and decays of the pH difference and membrane potential, followed using 9-aminoacridine and 8-anilinonaphthalene-1-sulfonate, respectively, as probes, showed a relationship between delta-micronH+ and the rate of ATP synthesis . These results demonstrate that purified TF0-F1 is itself a reversible H+-translocating ATPase of oxidative phosphorylation.

Genetics, 1977 May, 86(1), 103 - 12
Traumatic induction of early maturity in tetrahymena; Nanney DL et al.; Exposure of conjugating pairs of Tetrahymena thermophila to high temperature (37 degrees) during macronuclear development causes an abortion of many macronuclei, but it also often induces an early appearance of sexual maturity in clones completing macronuclear development . Lines become mature after about 15 cell divisions rather than after 50 or more cell divisions in untreated pairs . The phenotype resembles that associated with Em (early maturity) mutants but, because it is not transmitted to the progeny in the next generation, it must be considered a phenocopy . The hypothesis is developed that an early genotype-environment incompatibiltiy, whether associated with an abnormal genotype or an unusual environment, activates a shunt mechanism permitting the organisms to undertake quickly an ordinarily forbidden sexual lottery.

Can J Microbiol, 1977 May, 23(5), 510 - 7
Similar C-phycocyanins from two strains of thermotolerant cyanophyte Mastigocladus laminousus; Kao OH et al.; C-phycocyanin from two strains of the thermotolerant blue-green alga, Mastigocladus laminosus (NZ-DB2-m and I-30-m), that grow within different temperature ranges have been characterized with respect to aggregation, immunologic properties, subunit composition, and thermodenaturation . The critical thermal-denaturation temperature for phycocyanin from both strains of M . laminosus phycocyanin is 60 degrees C which is higher than that for mesophilic phyococyanin . Immunodiffusion studied have shown that these two strains of M . laminosus exhibit no antigenic differences and are closely related to the mesophilic Plectonema calothricoides and the thermophilic Synechococcus lividus (strains 3) . Neither phenol nor alpha-naphthol has any effect on phycocyanin aggregation in these two strains of M . laminosus . There is also no enhancement of formation of large aggregates at their elevated temperature of cultivation . Furthermore, the phycocyanin of both strains of M . laminosus does not demonstrate any large amount of 19S or higher aggregates at any pH value . These observations suggest that the mode of adaptation of M . laminosus phycocyanin to high temperature is differnet from the previously encountered . It is also important to note that phycocyanin is essentially unchanged whether it is extracted from the same strain, M . laminosus (NZ-DBS-m), grown at either 50 degrees C or 37 degrees C.

Proc Natl Acad Sci U S A, 1977 May, 74(5), 2045 - 9
Direction of active sliding of microtubules in Tetrahymena cilia; Sale WS et al.; Axonemes of protozoan (Tetrahymena thermophila BIII) cilia, isolated by the dibucaine method, were treated briefly with trypsin after removal of the ciliary membranes by treatment with Triton X-100 . After attachment to polylysine-coated surfaces, the partially digested axonemes remained mainly intact cylinders . Such attached axonemes can be treated with ATP, which induces microtubles sliding . ATP-treated preparations showed disrupted axonemes in which doublets had telescoped out of the original cylinders . These could be captured in place for electron microscopy after critical point drying . Images of this type were used to determine relative movement between adjacent doublet microtubules . Each doublet actively slid relative to its neighbors in a single direction, in which the polarity of force generation of the dynein arms was from base to tip.

Fed Proc, 1977 May, 36(6), 1815 - 8
Proton translocation by ATPase and bacteriorhodopsin; Kagawa Y et al.; Stable membrane proteins and lipids are convenient to study biomembranes . Two stable proton translocating proteins were purified and reconstituted into vesicles capable of proton translocation . One was a thermostable ATPase (TF0-F1) of thermophilic bacterium PS3 and the other was rhodopsin of Halobacterium halobium . TF0-F1 was composed of a proton pump moiety (TF1) and a proton channel moiety (TF0) . TF1 was the first membrane ATPase which was crystallized and reconstituted from its five polypeptides . Like TF0 and TF1, the rhodopsin in purple membrane was highly stable against dissociating agents, acids and alkali . Phospholipids of these biomembranes were also stable and contained no unsaturated fatty acyl groups . The molecular species of the phospholipids of PS3 were determined by mass chromatography . Measurements were made of the difference in electrochemical potential of protons (deltamicronH+) across the membrane of the reconstituted vesicles . The deltamicronH+ attained was 312 mV in TF0-F1 vesciles and was 230 mV in the rhodopsin vesicles . To conclude that electron transport components are not necessary for ATP synthesis in energy yielding biomembranes, two experiments were performed: The ATP synthesis was observed i) on acid-base treatment of TF0-F1 vesicles, and ii) on illumination of the rhodopsin-TF0-F1 vesicles.

J Biochem (Tokyo), 1977 Apr, 81(4), 1161 - 5
Determination of molecular species of phospholipids from thermophilic bacterium PS3 by mass chromatography; Kagawa Y et al.; The molecular species of phosphatidylethanolamine, phosphatidyglycerol and cardiolipin of thermophilic bacterium PS3 were determined by a new and simple mass chromatographic technique . The fatty acyl groups of these compounds, unlike those of usual phospholipids, were all saturated and were mixtures of even and odd numbered carbon chains; their {M-CnH2n+1-COOCH2}+ and {M-Cn+1H2n+3COO}+ peaks overlapped . Thus, the acetyldiglycerides derived from these phospholipids were subjected to a combination of mass chromatography using chemical ionization with NH3 and treatment of the phospholipids with phospholipase A2 {EC 3.1.1.4} . These three phospholipids had similar patterns of diglycerides . Expressed as R1 : R2, %, they were found to be 14:14,0-1%, 15:14,0-4%, 15:15, 22-23%, 16:15, 16-20%, 17:15, 28-32%, 17:16, 6-9%, 18:15, 7-13%, 17:17, 3-6%, and 18:16, 0-1% . These results should be of significance in connection with the high stability of the biomembranes of this bacterium.

J Biochem (Tokyo), 1977 Apr, 81(4), 1107 - 13
Biochemical studies on an acidophilic, thermophilic bacterium, Bacillus acidocaldarius: isolation of bacteria, intracellular pH, and stabilities of biopolymers; Oshima T et al.; Acidophilic, thermophilic bacteria were isolated from Japanese acidic hot springs . They were spore-forming rods, identified as Bacillus acidocaldarius . DNA extracted from these acido-thermophiles showed no abnormality in chemical structure; it was instantly denatured and gradually decomposed giving rise to apurinic acid in a hot acid environment milder than the optimal conditions for the growth of the acido-thermophiles . Glyceraldehyde-3-phosphate dehydrogenase extracted from B . acidocaldarius was not active at pH 5 or less, and was resistant to heat at neutral but not acid pH . The intracellular pH was computed to be neutral by using dimethyl-2,4-oxazolidinedione . When uncouplers or inhibitors of respiration were added to the cells suspended in hot acid solution, the estimated pH was not changed and glyceraldehyde-3-phosphate dehydrogenase in the cells was not denatured . These results suggest that the cytoplasm of B . acidocaldarius is a hot neutral environment, and that a pH gradient across the cell envelope can be maintained even when oxidative phosphorylation or respiration is inhibited.

Biochim Biophys Acta, 1977 Mar 29, 497(1), 112 - 21
Relationship between culture density and catabolite repression of an inducible aliphatic amidase in a thermophilic bacillus; Thalenfeld B et al.; A direct correlation between the absorbance of a thermophilic bacillus and specific amidase activity was observed, which was found to depend on the cell density of the culture rather than on the time of contact of the culture with the inducer . Dilution of high density cultures caused the specific amidase activity to decrease . Environmental factors such as pH, concentration of inducer or degree of aeration, and level of NH+4 and glutamate had no effect on amidase synthesis . The decrease in amidase activity upon dilution could not be ascribed to destruction by oxygen or by inactivation or decay . Several lines of evidence suggest that catabolite repression is responsible for the phenomenon described . Succinate-grown cultures gave a stronger dilution effect thatn glutamate-grown cells . The mutant strain E-21, relatively resistant to catabolite repression, did not show the characteristic dilution effect nor the direct correlation between absorbance and specific amidase activity.

Chromosoma, 1977 Mar 7, 60(1), 49 - 57
Cytogenetics of triplet conjugation in Tetrahymena: origin of haploid and triploid clones; Preparata RM et al.; Triplet conjugation occurs frequently when three different mating types of T . thermophila are mixed under appropriate conditions . Genetic studies show that fertilization is usually tripolar, with each conjugant contributing a pronucleus to one mate and receiving a pronucleus from the other . Triplets are less common and less stable when only two mating types are mixed, and nuclear exchange is not always reciprocal . Some conjugants contribute a pronucleus but do not receive one in return . Others receive two pronuclei instead of one . Such triplets characteristically yield one haploid, one diploid and one triploid exconjugant, all of which are able to establish vigorous clones.

Biochim Biophys Acta, 1977 Mar 2, 475(1), 32 - 41
DNA polymerase from mesophilic and thermophilic bacteria . III . Lack of fidelity in the replication of synthetic polydeoxyribonucleotides by DNA polymerase from Bacillus licheniformis and Bacillus stearothermophilus; Stenesh J et al.; 1 . DNA polymerase from the mesophile Bacillus licheniformis and the thermophile Bacillus stearothermophilus has been used to study the replication of poly(dA-dT)-poly(dA-dT) and poly(dC)-poly(dG) templates at 37, 45, and 55 degrees C . 2 . Incorporation of non-complementary deoxyribonucleoside triphosphates (misincorporation) occurred with both enzymes and both templates . Non-specific incorporation (de novo polynucleotide synthesis, random attachment to existing strands, and tritium exchange of nucleotides) accounted for, at most, a small fraction of the total observed misincorporation . The error rates at 37 degrees C for the complete system were as follows:: B . licheniformis: dATP, 1/61; dCTP, 1/830; dGTP, 1/360; dTTP, 1/65; B . stearothermophilus: dATP, 1/68; dCTP, 1/1430; dGTP, 1/440; dTTP, 1/67 . For both organisms, the error rate for dCTP and dGTP was independent of incubation temperature; the error rate for dATP and dTTP was 5-50-fold greater than that for dCTP or dGTP and increased significantly from 37 to 55 degrees C . 3 . The ratio of dATP to dTTP incorporation with the poly(dA-dT)-poly-(dA-dT) template was independent of temperature and close to unity . The ratio of dCTP to dGTP incorporation with the poly(dC)-poly(dG) template decreased from approx . 0.2 to 0.05 for the mesophile and from approx . 0.06 to 0.03 for the thermophile as the temperature increased from 37 to 55 degrees C.

Poult Sci, 1977 Mar, 56(2), 653 - 61
Growth of fungi in broiler houses; Bacon CW et al.; The growth of fungi in broiler houses with and without histories of problems of unknown etiology or of mycotoxicoses was studied . Fungal growth data were obtained by the strip bait method and results expressed as frequency and relative density . The frequency and relative density of fungal growth of several species were higher in problem houses than in those without problems . In problem houses many known toxic fungi and several fungi of unknown toxicity were found growing, increasing in density and frequency during the rearing period . Houses without problems (control houses) were dominated throughout the rearing period by the growth of Scopulariopsis brevicaulis . This fungus grew infrequently and in low relative density in problem houses . There was no correlation between species composition and litter removal, pH, and moisture in problem and control houses . Fungi did not grow in inside feed hoppers or in dust on rafters in the broiler houses . One thermotolerant and two thermophilic species of fungi were found growing in outside feed bins . Three generalized fungal growth patterns in litter are described and theoretical consideration of environmental factors on these patterns are discussed.

J Lab Clin Med, 1977 Mar, 89(3), 533 - 9
Indirect immunofluorescent detection of antibodies against thermophilic actinomycetes in patients with hypersensitivity pneumonitis; Kurup VP et al.; With slide culture preparations of Micropolyspora faeni, Thermoactinomyces candidus, and Thermoactinomyces vulgaris used as antigens, 59 sera from patients with hypersensitivity pneumonitis and 23 sera from normal controls were studied for the presence of specific antibodies by indirect immunofluorescence and immunodiffusion methods . Of the 82 sera, 31 showed precipitins to M . faeni, five to T . candidus, and four to T . vulgaris . A group of 19 sera positive to Aspergillus antigens showed questionable reactions to thermophilic actinomycetes . The 23 sera from normal individuals were negative for specific antibodies when tested by immunodiffusion . In indirect immunofluorescence studies, we found that IgG antibodies in high titers are present only in the precipitin-positive sera whereas other antibody classes either are negative or failed to show consistent differences in titers of the positive, questionable, and control groups . The titer of IgG antibodies varied from 0 to 1:20 in the normal control group, 0 to 1:40 to 1:640 in the precipitin-positive group.

Nucleic Acids Res, 1977 Mar, 4(3), 585 - 93
Proton magnetic resonance spectra of tRNA-Met-f from Thermus thermophilus; Kyogoku Y et al.; 220 MHz proton magnetic resonance spectra of tRNAs in bulk and tRNA-Met-f from Thermus thermophilus have been measured and compared with those of tRNAs from E . coli . Temperature dependences and chemical shift positions of the bulk tRNAs are well explained by the difference in their GC contents . It is known that the base sequence of the double helical regions in the cloverleaf structure of T . thermophilus tRNA-Met-f is different from that of E . coli tRNA-Met-f only at two positions in TpsiCarm; one more C:G pair is contained instead of a U:G pair of E . coli tRNA-Met-f and a C:G pair of E . coli is replaced by a G:C pair . In spite of the resembrance in the base sequences, nmr patterns around 13 ppm are fairly different from each other . The difference is discussed in relation with their tertiary structures and with the origin of chemical shift displacements.

Mikrobiologiia, 1977 Mar-Apr, 46(2), 342 - 5
{Several cytologic features of the thermophilic bacteria Thermus flavus and Thermus ruber}; Loginova LG et al.; The extreme thermophilic bacterium Thermus flavus 71 and the obligate thermophilic bacterium Thermus ruber 40 have been isolated from hot springs of Kamchatka and Tadjikistan, and their fine structure has been studied . This cells are similar in structure to gram-negative bacteria, they do not form spores or flagella, and their cell walls and cytoplasmic membrane are clearly visible under the electron microscope since the cells of T . flavus contain a yellow carotenoid pigment and the cells of T . rubber contain a bright-red carotenoid pigment . The Thermus genus is characterized by a wavy structure of the outer layer of the cell wall caused by numerous invaginations which ensure a close contact between the outer and inner layers of the cell wall.

Proc Natl Acad Sci U S A, 1977 Mar, 74(3), 936 - 40
Reconstitution of thermostable ATPase capable of energy coupling from its purified subunits; Yoshida M et al.; Purified dicyclohexylcarbodiimide-sensitive ATPase (TF0-F1) from thermophilic bacterium PS3 is composed of a water soluble part with ATP hydrolytic activity (TF1) and a water insoluble moiety (TF0) . All of the five subunits (alpha, beta, gamma, delta, and epsilon) of TF1 were isolated . TF1 was reconstituted from the five subunits, which catalyzed an ATP-32Pi exchange and an ATP-driven enhancement of fluorescence of 1-anilinonaphthalene-8-sulfonate, when adsorbed on proteoliposome inlaid with TF0 (TF3-vesicles) . Subunit epsilon and/or delta became firmly bound to TF0-vesicles and there was no preferential sequence in the binding . Both subunits were required for binding of the remaining subunits of TF1 to TF0-vesicles, but they did not modify the high H+ -permeability of TF0-vesicles . The addition of gamma but they did not modify the high H+-permeability of TFO-vesicles . The addition of gamma subunit together with epsilon and delta subunits caused a marked decrease of H+ -permeability of TF0-vesicles, similar to that induced by TF1 . We conclude tentatively that the epsilon and delta subunits connect TF0 and the other subunits forming a part of a proton pathway, gamma is a gate of proton flow coupled to ATP hydrolysis (or synthesis), and alpha and beta subunits contain the active site for energy transformation . A possible model of subunit structure of TF1 is proposed.

Eur J Biochem, 1977 Mar 1, 73(2), 373 - 81
Superoxide dismutase from Thermus aquaticus . Isolation and characterisation of manganese and apo enzymes; Sato S et al.; Superoxide dismutase has been isolated and characterised from the extreme thermophile Thermus aquaticus . The pure enzyme is a reddish-purple manganese-containing protein with a molecular weight of approximately 80000 +/- 5000 . Combination of gel electrophoresis in dodecylsulphate and amino acid analysis shows that it is composed of four identical subunit polypeptide chains consisting of approximately 186 amino acids . The tetrameric protein contains two atoms of manganese . A stable manganese-free apoprotein has been prepared by treatment with EDTA in 8 M urea at acidic pH . The apoprotein regains the tetrameric structure in the absence of manganese but is inactive . Reconstitution of active Mn-enzyme was achieved byaddition of Mm2+ apoprotein in 8 M urea at acid pH . Reconstitution was monitored by absorption spectroscopy, manganese analysis and regain of activity and by these criteria the reconstituted enyzme with two atoms Mn per mole is indistinguishable from the native enzyme . The enhanced stability of the thermophile apoenzyme and Mn-enzyme is of advantage for studies of the structure and mechanism of action of superoxide dismutase . The N-terminal amino acid sequence to the 40th residue of the submit was determined by automated Edman degradation . The sequence has a close resemblance to that of the dimeric Mn-enzyme from another thermophile, Bacillus stearothermophilus.

J Bacteriol, 1977 Feb, 129(2), 690 - 7
Purification and properties of homoprotocatechuate 2,3-dioxygenase from Bacillus stearothermophilus; Jamaluddin MP; The enzyme 3,4-dihydroxyphenylacetate:oxygen 2,3-oxidoreductase (decyclizing) (homoprotocatechuate 2,3-dioxygenase) was purified from the thermophilic organism Bacillus stearothermophilus, grown with j-hydroxyphenylacetic acid as a source of carbon . The enzyme appeared to be homogeneous as judged by disc-gel electrophoresis and sedimentation equilibrium measurements . The average molecular weight determined by three independent procedures was 106,000; the protein was globular and was dissociated in sodium dodecyl sulfate to give a species of molecular weight 33,000 to 35,000 . The enzyme was fairly stable on heating and showed maximal activity at about 57 degrees C . An Arrhenius plot of Km for homoprotocatechuate was concave upward, with a break at 32 degrees C; an increase in delta H above this temperature was compensated by lower values of --delta S . Several properties of this enzyme are contrasted with those reported for homoprotocatechuate 2,3-dioxygenase purified by other workers from Pseudomonas ovalis.

Appl Environ Microbiol, 1977 Feb, 33(2), 298 - 307
Thermophilic methane production from cattle waste; Varel VH et al.; Methane production from waste of cattle fed a finishing diet was investigated, using four 3-liter-working volume anaerobic digestors at 60 degrees C . At 55 degrees C a start-up culture, in which waste was the only source of bacteria, was generated within 8 days and readily adapted to 60 degrees C, where efficiency of methanogenesis was greater . Increasing the temperature from 60 to 65 degrees C tended to drastically lower efficiency . When feed concentrations of volatile solids (VS, organic matter) were increased in steps of 2% after holding for 1 months at a given concentration, the maximum concentrations for efficient fermentation were 8.2, 10.0, 11.6, and 11.6% for the retention times (RT) of 3, 6, 9, and 12 days, respectively . The VS destructions for these and lower feed concentrations were 31 to 37, 36 to 40, 47 to 49 and 51 to 53% for the 3-, 6-, 9-, and 12-day RT digestors, respectively, and the corresponding methane production rates were about 0.16, 0.18, 0.20, and 0.22 liters/day per g of VS in the feed . Gas contained 52 to 57% methane . At the above RT and feed concentrations, alkalinity rose to 5,000 to 7,700 mg of CaCo3 per liter (pH to 7.5 to 7.8), NH3 plus NH4+ to 64 to 90 mM, and total volatile acids to 850 to 2,050 mg/liter as acetate . The 3-day RT digestor was quite stable up to 8.2% feed VS and at this feed concentration produced methane at the very high rate of 4.5 liters/day per liter of digestor . Increasing the percentage of feed VS beyond those values indicated above resulted in greatly decreased organic matter destruction and methane production, variable decrease in pH, and increased alkalinity, ammonia, and total volatile acid concentrations, with propionate being the first to accumulate in large amounts . In a second experiment with another lot of waste, the results were similar . These studies indicate that loading rates can be much higher than those previously thought useful for maximizing methanogenesis from cattle waste.

Proc Natl Acad Sci U S A, 1977 Feb, 74(2), 542 - 6
A thermostable sequence-specific endonuclease from Thermus aquaticus; Sato S et al.; A sequence-specific endonuclease, Taq I, of novel specificity has been partially purified from an extreme thermophile, Thermus aquaticus . The enzyme cleaves bacteriophage lambda DNA at many (greater than 30) sites and bacteriophage psiX174 RF DNA at 10 sites . The enzyme is active at temperatures up to 70 degrees . The cleavage sites on psiX174 RF DNA have been mapped . The sequence recognized and cleaved by Taq I has been shown to be the symmetrical tetranucleotide: (formula: see text).

Infect Immun, 1977 Feb, 15(2), 608 - 13
Extracellular antigens of Micropolyspora faeni grown in synthetic medium; Kurup VP et al.; Extracellular antigens were prepared by growing Micropolyspora faeni in a new synthetic medium (SMSL), and this antigenic preparation was compared with antigens obtained by the double-dialysis method . These two preparations were tested against pooled sera from farmer's lung patients and rabbits immunized with these antigens by the agar-gel double-diffusion, immunoelectrophoresis, and two-dimensional immunoelectrophoresis methods . The immunological cross-reactivity between M . faeni strains of diverse origin and between other species of thermophilic actinomycetes associated with hypersensitivity pneumonitis was also investigated . The usefulness of two-dimensional immunoelectrophoresis in the comparison and standardization of antigens is discussed.

J Biochem (Tokyo), 1977 Feb, 81(2), 519 - 28
Reconstitution of vesicles capable of energy transformation from phospholipids and adenosine triphosphatase of a thermophilic bacterium; Sone N et al.; 1 . A stable ATPase {EC 3.6.1.3} complex (TF0-F1) from the thermophilic bacterium PS3 was reconstituted into vesicles capable of energy transformation,measured as ATP-dependent enhancement of fluorescence of 8-anilinonoaphthalene-1-sulfonate . 2 . The factors necessary for obtaining highly active vesicles were investigated . Cholate and deoxycholate were both required for solubilization of TF0-F1 and P-lipids, and removal of the detergents by dialysis resulted in vesicle formation . Medium of around pH 8 and low ionic strength containing 2.5 mM MgSO4 was found suitable for dialysis . The optimal temperature for reconstitution was 30 degrees with soybean P-lipids and 45 degree with PS3 P-lipids . The optimal ratio of protein to lipid was about 1/50 . 3 . The vesicles obtained under these conditions were mainly 100-200 nm in diameter, covered with 9.5 nm spheres, and had a bouyant density of 1.06 in sucrose andan internal volume of about 0.5 mul per mg of P-lipids.

Rev Asoc Argent Microbiol, 1977 Jan-Apr, 9(1), 17 - 21
{Description of Streptomyces malvinense nov . sp . isolated from the soil of Islas Malvinas}; Cercos AP; In the course of a study on antibiotic activity of microorganisms isolated from soil samples from Port Stanley, Islas Malvinas, Argentina, four thermophilic Actinomycetes were sudied . The four strains had the same morphological and physiological characteristics . The new species belonged to Seccion Rectiflexibilis genus Streptomyces and had the following characteristics: sporophores straight, round spores of 1,5 or 2 millimicron in diameter . The colonies in agar meat peptone were round, flat, mealy, with fimbriate edges, odorless, friable, easy to emulsify and the aerial mycelium was white to light gray . The sporulation was rapid (less of 24 hours at 50 degrees C), the microorganisms grew in media with organic nitrogen and his ability to utilize carbon was scarce, only a trace of growth was detected with manitol (20 days at 50 degrees C) . No growth was observed in gliceril-asparagine agar and the utilization of glucose was scarce or absent . It produced coagulation and peptonization of milk and liquifaction of gelatin . Neither was starch hidrolized nor nitrate reduced . The optimun growth temperature was between 45 - 60 degrees C . Very resistant to high temperatures having a thermal death point of more that 2 hours at 100 degrees C . It produced SH2.

Int J Pept Protein Res, 1977, 10(1), 39 - 50
Comparison of proteins from thermophilic and nonthermophilic sources in terms of structural parameters inferred from amino acid composition; Singleton R Jr et al.; The amino acid composition of 14 different proteins from thermophilic bacteria were compiled along with the amino acid compositions of 56 corresponding proteins from nonthermophilic sources . A comparison was made between proteins serving the same catalytic function, and significant differences in composition were noted for those proteins from thermophilic bacteria . However, no consistent pattern was evident and the differences were often small . The two data pools were treated as two distinct classes and a thermophilic versus non-thermophilic comparison of amino acid composition was made using the Student's t-test . Significant differences in composition were found for Asx (sum of Asp and Asn, if known), Ser, and Arg . Both classes of data have similar standard deviations for the mean of any single amino acid, suggesting a similar tolerance of variation in the two classes of proteins . This would argue against the hypothesis that thermophiles exhibit a greater frequency of errors in protein synthesis . The amino acid compositions were used to calculate structural parameters (% helix, % beta, % turn, hydrophobicity, and melting temperatures) for the two classes of proteins . Of these, only the predicted % beta content was significantly lower for proteins of thermophilic origin . No differences in hydrophobicity or predicted melting temperature were observed for the two classes of proteins . This study supports the hypothesis that while small differences may occur in the amino acid composition of thermophilic proteins, they are quite varied and often are very subtle.

Folia Parasitol (Praha), 1977, 24(2), 149 - 60
Influence of microclimate on the life cycle of the common tick Ixodes ricinus (L.) in an open area in comparison with forest habitats; Daniel M et al.; Under conditions of the South-Moravian region of Pannonian climate (Valtice near Breclav), the life cycle of the common tick Ixodes ricinus (L.) was studied and a continous recording of main elements of microclimate (temperature and humidity) was carried out in an open grassy area . Simultaneously the process of hibernation was studied in four soil layers (surface, depths of 10, 20 and 30 cm) . Observations were assessed by mathematiccal-statistical tests and compared with the results obtained by the same methods in the forest biotope (tipe of thermophilic oak forest)and in the ecotone of forest margin (Daniel et al . 1976) . The ticks are able to complete the developmental cycle also in the open grassy areas, where during the vegetation period the development proceeds more quickly than in the forest but with considerably higher losses . In the discussion the conclusions are compared with literary data from other parts of Czechoslovakia.

Zentralbl Bakteriol Parasitenkd Infektionskr Hyg, 1977, 132(2), 143 - 7
Studies on the formation of alpha-amylase by Thermomonospora vulgaris; Allam AM et al.; Conditions affecting the formation of alpha-amylase by static cultures of the thermophilic actinomycete Thermomonospora vulgaris were studied . The organism failed to grow under submerged culture conditions or when the culture medium was devoid of CaCO3-alpha-Amylase was produced during the logarithmic phase of growth and maximum yield was obtained after 3 to 9 days of incubation . Growth and amylase formation took place only in a range from 45 degrees to 55 degrees C; optimum temperature was 55 degrees C . Of the tested carbon sources only starch induced enzyme formation . Maximum enzyme yield was obtained when starch concentration of the medium was 2% and when ammonium citrate served as a nitrogen source . Crushed clay pots could substitute for CaCO3 of the medium, but growth and amylase yield were less.

Mikrobiologiia, 1977 Jan-Feb, 46(1), 38 - 40
{Influence of carbon sources on the development of Thermus ruber at different temperatures}; Loginova LG et al.; The obligate thermophilic bacterium Thermus ruber was found to grow better at 55-65 degrees C on media containing organic acids rather than sugars as a sole source of carbon . In these conditions, the morphology of the cells changes: they are rod-shaped whereas, during growth on lactose which is the best carbon source as compared to other sugars, the cells have the shape of rods in the first hours of growth but later acquire the shape of long threads . Malate and pyruvate are the best carbon sources at 60-65 degrees C of growth; the biomass accumulated in this case is 1.5 times greater than on the medium containing lactose . Apparently, salts of organic acids, being more oxidized substrates as compared to sugars, are easier assimilated by the bacterium at higher temperatures when oxygen solubility in the medium decreases.

Int J Pept Protein Res, 1977, 9(4), 258 - 68
Thermophilic aminopeptidase . IV . Cooperative effects in ANS binding by the thermophilic aminopeptidase I from B . stearothermophilus; Deranleau DA et al.; Aminopeptidase I is a membrane-bound metalloenzyme isolated from B . stearothermophilus which is thermostable and requires Co2+ for activity . The Co, Zn, and metal-free enzyme were titrated with ANS, and cooperative binding was noted with the active Co enzyme but not with the Zn (5% active) or apo (inactive) enzymes . There are a number of sites for ANS on each of the three enzyme forms and the agreement between the association constants of the Zn enzyme (identical and independent sites) and the non-cooperative sites of the Co enzyme suggest that these sites are intrinsically similar . However, binding to the first of these sites in the Co enzyme triggers a cooperative binding of a second molecule of ANS, and this cooperative binding is related to a concomitant decrease in enzymatic activity . The correspondence can be shown by comparison of the inhibition constant for the hydrolysis of Gly-Leu-Tyr (Ki-1=13,300 cm3/mmol) and the association constant for the cooperating site (12,500 cm3/mmol) . The significance of these observations is discussed in terms of the nature of the binding sites and the possible consequences of the interactions on the regulation of aminopeptidase I activity.

Cell Biol Int Rep, 1977 Jan, 1(1), 69 - 73
Dibucaine-induced synchronous mucocyst secretion in Tetrahymena; Satir B; Synchronous secretion of all available mature mucocysts was induced in late log phase cultures of Tetrahymena thermophilia (B III) by the local anaesthetic dibucaine . No assembled fusion rosettes were seen within the plasma membrane after release until 2-3 hrs of regrowth, thus proving that the rosettes are not permanent sites within the plasma membrane but have to be reassembled each time for a new fusion event to occur . Concomitant with the reappearance of assembled fusion rosettes, the cell cytoplasm fills up with precursors of new mucocysts thus linking the two events together.

J Bacteriol, 1977 Jan, 129(1), 501 - 15
Electron microscope histochemical localization of alkaline phosphatase(s) in Bacillus licheniformis; McNicholas JM et al.; Sites of alkaline phosphatase (APase) activity in a facultative thermophilic strain of Bacillus licheniformis MC14 have been localized by electron microscope histochemistry, using a lead capture method . The effects of 3% glutaraldehyde and 3.0 mM lead on APase activity were investigated, and these compounds were found to significantly inhibit enzyme activity, 68 and 18%, respectively . A number of parameters were varied in studies to localize APase activity, including: growth temperature (55 and 37 degrees C); substrate concentration in the histochemical mixture (0.06, 0.15, 0.30, 1.00 mM); fixatives; protoplast preparations and whole cells; phosphate-repressed and -derepressed cells; and age of vegetative cells (mid-log and late log) . These variations affected the number but not the location of lead phosphate deposits, which appeared at discrete sites along the inner side of the cytoplasmic membrane . Control cells incubated in histochemical mixtures lacking substrate, lead, or both exhibited no lead phosphate depositis . The histochemical localization at membrane sites correlated well with biochemical localization data, which indicated that greater than 80% of the APase activity was associated with the membrane fraction in logarithmically growing cells.

J Supramol Struct, 1977, 6(1), 77 - 84
Isolation of the alanine carrier from the membranes of a thermophilic bacterium and its reconstitution into vesicles capable of transport; Hirata H et al.; A carrier protein mediating alanine transport was purified from the membranes of the thermophilic bacterium PS3, by ion exchange chromatography in the presence of both Triton X-100 and urea . The alanine carrier was recovered in the nonadsorbed fraction from either DEAE- or CM-cellulose columns, suggesting that its isoelectric point was in the neutral pH region . The final preparation contained virtually no electron transfer components, ATPase, or NADH dehydrogenase . Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the final preparation consisted of two major protein components with molecular weights of 36,000 and 9,400 . Active transport of alanine after incorporation of the alanine carrier into reconstituted proteoliposomes was driven not only by an artificial membrane potential generated by potassium ion diffusion via valinomycin but also by mitochondrial cytochrome oxidase incorporated into the same liposomes and supplemented with both cytochrome c and ascorbic acid . The membrane-integrated portion (TFo) of the ATPase complex uncoupled alanine transport by conducting protons across the membrane.

Acta Microbiol Pol, 1977, 26(2), 213 - 20
Development of some groups of microorganisms in liquid cattle manure and farmyard manure during their fermentation; Martyniuk S et al.; The development of meso- and thermophilic microorganisms proceeded more strongly in the examined farmyard manure than in the liquid cattle manure . The most vigorous proliferation of mesophilic bacteria and fungi was found in both dungs during the first weeks of fermentation . Relatively greater numbers of mesophilic actinomycetes were noticed in the later experimental period . Compared to mesophilic microoganisms, thermophilic ones were less numerous and quantitatively changeable in the tested dungs . In contrast to the farmyard manure, in the liqued dung thermophilic actinomycetes did not occur at all . After 3 monts of the experiment the Coliform titre decreased to 101 in the liquid manure, but to 104 in the farmyard manure.

Int J Pept Protein Res, 1977, 9(5), 329 - 39
Fluorescent labelling of 6-phosphogluconate dehydrogenase from Bacillus stearothermophilus; Fontana A et al.; The thermophilic 6-phosphogluconate dehydrogenase from Bacillus stearothermophilus was inhibited upon specific modification of the -SH group of cysteine residues by 7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole (NBD-Cl) at pH 7.0 . By using 20-100-fold molar excess of NBD-CL the reaction occurs slowly at pH 7.0 as a first order process . Partial protection from inactivation was observed when the substrate 6-phosphogluconate or the coenzyme NADP was added to the reaction mixture . Complete inactivation was achieved upon modification of 1.9 of the six cysteine residues per mole of enzyme, which corresponds to nearly one residue per enzyme subunit . Circular dichroism measurements suggest that the gross structure of the protein molecule is practically unchanged upon reaction of the enzyme with NBD-Cl . Melting profile experiments revealed a single transition occurring at about 65 degrees C . Analogously, the profile of intensity of the fluorescence emission at 520 nm of the enzyme-bound S-NBD groups versus temperature indicated a midpoint of transition near 65 degrees C . Since this melting temperature corresponds closely to that observed with the native enzyme, these results would indicate that the molecular organizations of the native and modified enzyme are similar and stabilized by similar interactions within the polypeptide chain.

J Membr Biol, 1976 Dec 28, 30(2), 121 - 34
Electrochemical potential of protons in vesicles reconstituted from purified, proton-translocating adenosine triphosphatase; Sone N et al.; Measurements were made of the difference in the electrochemical potential of protons (delta-mu H+) across the membrane of vesicles restituted from the ATPase complex (TF0.F1) purified from a thermophilic bacterium and P-lipids . Two fluorescent dyes, anilinonaphthalene sulfonate (ANS) and 9-aminoacridine (9AA) were used as probes for measuring the membrane potential (delta psi) and pH difference across the membrane (delta pH), respectively . In the presence of Tris buffer the maximal delta psi ans no delta pH were produced, while in the presence of the permeant anion NO-3 the maximal delta pH and a low delta psi were produced by the addition of ATP . When thATP concentration was 0.24 mm, the delta psi was 140-150 mV (positive inside) in Tris buffer, and the delta pH was 2.9-3.5 units (acidic inside) in the presence of NO-3 . Addition of a saturating amount of ATP produced somewhat larger delta psi and delta pH values, and the delta -muH+attained was about 310mV . By trapping pH indicators in the vesicles during their reconstitution it was found that the pH inside the vesicles was pH 4-5 during ATP hydrolysis . The effects of energy transfer inhibitors, uncouplers, ionophores, and permeant anions on these vesicles were studied.

J Cell Physiol, 1976 Dec, 89(4), 569 - 73
Transport of nutrients by a thermophilic bacterium--reconstruction of vesicles from crystalline ATPase or solubilized alanine carrier; Kagawa Y; A strain of aerobic thermophilic bacteria was selected in order to purify highly stable membrane proteins and no reconstitute proteoliposomes capable of transporting nutrients from them . These proteins responsible for the transport could be divided into (1) proteins which supply energy to the transporting system, and (2) specific nutrient carriers driven by the energy . The former included a stable ATPase (TF1) and a lipoprotein TF0) which rendered TF1 sensitive to energy transfer inhibitors . The complex of TF0 anlysis of ATP . And one of the latter reported in this paper was alanine carrier protein which was driven by proton movement . TF1 was the first crystallized ATPase in biomembranes, and was reconstituted from its five different polypeptides, two of which were necessary for ATPase activity and four of which, for proton translocation . Purification of alanine carrier and reconstitution of proteoliposomes capable of alanine accumulation were also demonstrated.

Appl Environ Microbiol, 1976 Dec, 32(6), 747 - 52
Production of an extracellular maltase by thermophilic Bacillus sp . KP 1035; Suzuki Y et al.; Production of extracellular maltase was studied with thermophilic Bacillus sp . KP 1035, which was selected as the organism producing the highest levels of maltase . The final enzyme yield was increased by maltose, peptone, and yeast extract but reduced by succinate and fumarate . Maximum enzyme production was achieved at 55 degrees C and at an initial pH of 6.2 to 7.0 on a medium containing 0.3% maltose, 1% peptone, 0.1% meat extract, 0.3% yeast extract, 0.3% KH2PO4, and 0.1% KH2PO4 . Maltase was synthesized in cytoplasm and accumulated as a large pool during the logarithmic growth phase, which preceded sporulation . At the end of this phase, the enzyme appeared in the culture broth, and its accumulation increased in parallel with a rise in the extracellular protein level . Maltase was stable for 24 h at 60 degrees C over a pH range of 5.6 to 9.0 and retained 95% of the original activity after treatment for 20 min at 70 degrees C at pH 6.8.

Biochim Biophys Acta, 1976 Nov 18, 451(1), 278 - 86
Thermoplasma acidophilum: intracellular pH and potassium concentration; Searcy DG; Thermoplasma acidophilum is a free-living thermophilic mycoplasma . Although the organism lacks a cell wall, it can grow in medium as dilute as 66 mosM . The intracellular K+ concentration can be as low as 17 mM, but varies according to the osmolality of the culture medium . The internal pH can be measured by taking advantage of the fact that T . acidophilum undergoes lysis when the pH is adjusted to neutrality . Thus, by appropriate analysis of titration curves, it is possible to conclude that the internal pH is near 5.5 . This result was confirmed by a second type of experiment in which the internal pH was analyzed by rupturing the cells in a French Pressure Cell.

Biokhimiia, 1976 Nov, 41(11), 2001 - 6
{Multiplicity of molecular forms and thermostability of Torula thermophila UzPT-1 protease}; Karavaeva NN et al.; Alkaline protease preparations with different ratio of molecular forms are isolated from cultural medium of thermophilic fungi Torula thermophila UzPT-1 by means of protein fractionation with (NJ/)2SO4 and gel filtration through Sephadex G-75 . The enzyme preparations differ in their thermostability in water at 60 degrees C . High molecular weight oligomeric enzyme forms dissociate in water (at 2-4 degrees C) forming dimeric and monomeric forms . Disaggregation is accompanied by the change in the thermostability of the enzyme preparations . It is concluded that protease thermostability depends on the ratio of dimeric and monomeric forms of the preparation, and it is associated with the conformational state of the enzyme molecules, and it is associated with the conformational state of the enzyme molecules . Oligomeric forms do not dissociate in 1% sodium dodecylsulphate and in 6 M urea . Ca2+ produces dissociation of high molecular weight enzyme forms and the conformational transition into the thermostable state.

Mikrobiologiia, 1976 Nov-Dec, 45(6), 1087 - 91
{Thermophilic bacteria from hot springs of Kamchatka}; Loginova LG et al.; Spore-forming bacteria with the optimum growth temperature from 40 to 65 degrees C were isolated from hot springs of Kamchatka with the temperature 50-93 degrees C and pH 6.3-7.3: Bacillus stearothermophilus, B . brevis, B . subtilis, B . cereus, B . megaterium, and B . licheniformis . The bacterial have a high biochemical activity and play therefore an important role in decomposition of organic substances.

Biochim Biophys Acta, 1976 Oct 11, 445(3), 549 - 57
Regulatory control and function of alanine dehydrogenase from a thermophilic bacillus; Epstein I et al.; L-alanine dehydrogenase, (L-alanine:NAD+ oxidoreductase (deaminating), EC 1.4.1.1) synthesis in a thermophilic bacillus was found to be subjected to regulatory control . Addition of L- and D-alanine and L-serine to cultures growing in the presence of either succinate or pyruvate, induced an accelerated synthesis of the alanine dehydrogenase enzyme . Synthesis of the enzyme was dependent on the presence of inducer during growth and was arrested by addition of glucose . Catabolite repression by glucose was abolished by limiting the ammonium concentration during growth . The apparent Km values of the substrates involved in alanine dehydrogenase activity are as follows (M): NH4+, 4-10(-2); pyruvate, 5-10(-4); NADH, 6-10(-5); L-alanine, 3.1-10(-3) and NAD, 2-10(-4) . Alanine dehydrogenase activity was measurable at temperatures below the minimal growth temperature (at 25 degrees C) and the highest activity was found at 65 degrees C; heat denaturation occurred at 80 degrees C.

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