J Gen Microbiol, 1983 Apr, 129 (Pt 4), 1005 - 11 Isolation and growth of a Pseudomonas species that utilizes cyanide as a source of nitrogen; Harris R et al.; A simple method of isolating bacteria that utilize cyanide as a source of nitrogen for growth has been developed . This involved supplying hydrogen cyanide as a vapour to glucose-containing minimal-salts agar plates . The bacteria isolated were Gram-negative, oxidase-positive rods producing a fluorescent green pigment and were tentatively identified as strains of Pseudomonas fluorescens . Three organisms were studied further and shown to be P . fluorescens biotype II . One of these (NCIB 11764) was grown in a glucose-containing fed-batch culture with either NH4Cl or KCN as the limiting nutrient . Cyanide-grown bacteria produced stoichiometric amounts of ammonia from cyanide when pulsed with cyanide under aerobic conditions . Stimulation of oxygen uptake was seen on addition of cyanide to suspensions of cyanide-grown but not ammonia-grown bacteria. Nucleic Acids Res, 1983 Mar 11, 11(5), 1245 - 52 Sequences of the 5S rRNAs of Azotobacter vinelandii, Pseudomonas aeruginosa and Pseudomonas fluorescens with some notes on 5S RNA secondary structure; Dams E et al.; Recently published alignments of available 5 S rRNA sequences have shown that a rigid base pairing pattern, pointing to the existence of a universal five-helix secondary structure for all 5 S RNAs, can be superimposed on such alignments . For a few species, the alignment and the base pairing pattern show distortions with respect to the large majority of sequences . Their 5 S RNAs may form exceptional secondary structures, or there may just be errors in the published sequences . We have examined such a case, Pseudomonas fluorescens, and found the sequence to be in error . The corrected sequence, as well as those of the related species Azotobacter vinelandii and Pseudomonas aeruginosa, fit perfectly in the 5 S RNA sequence alignment and in the five-helix secondary structure model . There exists comparative evidence for the frequent presence of non-standard base pairs at several points of the 5 S RNA secondary structure. Biophys J, 1983 Mar, 41(3), 233 - 44 Conformational heterogeneity of the copper binding site in azurin . A time-resolved fluorescence study; Szabo AG et al.; Comparison of the fluorescence spectra and the effect of temperature on the quantum yields of fluorescence of Azurin (from Pseudomonas fluorescens ATCC-13525-2) and 3-methylindole (in methylcyclohexane solution) provides substantive evidence that the tryptophan residue in azurin is completely inaccessible to solvent molecules . The quantum yields of azurin (CuII), azurin (CuI), and apoazurin (lambda ex = 291 nm) were 0.052, 0.054, and 0.31, respectively . Other evidence indicates that there is no energy transfer from tyrosine to tryptophan in any of these proteins . The fluorescence decay behavior of each of the azurin samples was found to be invariant with emission wavelength . The fluorescences of azurin (CuII) and azurin (CuI) decay with dual exponential kinetics (tau 1 = 4.80 ns, tau 2 = 0.18 ns) while that of apoazurin obeys single exponential decay kinetics (tau = 4.90) . The ratio of pre-exponentials of azurin (CuII), alpha 1/alpha 2, is found to be 0.25, and this ratio increases to 0.36 on reduction to azurin (CuI) . The results are interpreted as originating from different interactions of the tryptophan with two conformers of the copper-ligand complex in azurin. Biochem Pharmacol, 1983 Feb 15, 32(4), 679 - 89 Inhibition of glutamate-aspartate transaminase by beta-methylene-DL-aspartate; Cooper AJ et al.; beta-Methylene-DL-aspartate, a new beta, gamma-unsaturated amino acid, is an irreversible inhibitor of soluble pig heart glutamate-aspartate transaminase (Ki approximately 3 mM with respect to the L-form; limiting rate constant for inactivation approximately 0.4 min-1) . The new amino acid is the most specific inhibitor of glutamate-aspartate transaminase thus far studied . It does not inactivate pig heart glutamate-alanine transaminase, soluble rat kidney glutamine transaminase K, gamma-aminobutyrate transaminase (from Pseudomonas fluorescens), glutamate decarboxylase (Escherichia coli), snake venom L-amino acid oxidase, or hog kidney D-amino acid oxidase . In addition, the following enzymes were not inhibited by beta-methylene-DL-aspartate in rat tissue homogenates: gamma-aminobutyrate transaminase (brain), tyrosine transaminase (liver), glutamine transaminase L (liver), asparagine, transaminase (liver), ornithine transaminase (liver) or branch-chain transaminase(s) (kidney) . Intraperitoneal injection of beta-methylene-DL-aspartate into mice decreased kidney and liver glutamate-aspartate transaminase activities but had no effect on liver glutamate-alanine transaminase activity. J Dairy Res, 1983 Feb, 50(1), 77 - 89 Isolation and some properties of extracellular heat-stable lipases from Pseudomonas fluorescens strain AFT 36; Fox PF et al.; Aeration increased the growth and lipase production in milk by Pseudomonas fluorescens strain AFT 36, isolated from refrigerated bulk milk . A heat-stable lipase was isolated from a shaken milk culture of this microorganism by DEAE-chromatography and gel filtration in Sepharose 6B . The lipase-rich fraction from DEAE cellulose contained 3 lipases that were separated by gel filtration; only the principal lipase, which represented approximately 71% of total lipolytic activity, was characterized . The purified enzyme showed maximum activity on tributyrin at pH 8.0 and 35 degrees C; it had a Km on tributyrin of 3.65 mM and was inhibited by concentrations of substrate greater than approximately 17 mM . The enzyme was very stable over the pH range 6-9; it was relatively heat-labile in phosphate buffer in the temperature range 60-80 degrees C, where it was stabilized significantly by Ca2+ . It was, however, very stable at 100-150 degrees C: the D values at 150 degrees C were approximately 22 s and 28 s in phosphate buffer and synthetic milk serum respectively; the corresponding Z values in the temperature range 100-150 degrees C were approximately 40 and approximately 42 degrees C and the Ea for inactivation were 7.65 X 10(4) J mol-1 and 6.97 X 10(4) J mol-1 respectively. Eur J Biochem, 1983 Feb 1, 130(2), 391 - 6 6-phospho-D-gluconate dehydrogenase from Pseudomonas fluorescens . Properties and subunit structure; Stournaras C et al.; 1 . The 6-phospho-D-gluconate dehydrogenase (decarboxylating) (EC 1.1.1.44) from Pseudomonas fluorescens, a B-side stereospecific enzyme, is active with both NAD+ and NADP+, having a specific activity of the homogeneous enzyme of 121 mumols NADH and 23 mumols NADPH, respectively, formed min-1 mg protein-1 . The pI of the native enzyme is 4.62, the pH optimum is about 8.2 . 2 . The molecular weight of the native enzyme has been determined to be 126000 by sedimentation equilibrium studies . The molecular weight of the polypeptide chains composing the enzyme has been found to be 32000 by dodecylsulfate/polyacrylamide gel electrophoresis and 31000 by sedimentation equilibrium studies in presence of 6 M guanidine hydrochloride . The native enzyme is composed of four polypeptide chains . 3 . Reacting enzyme centrifugation studies gave at pH 8.2 a sedimentation coefficient s20, w of 8.04 S and a diffusion coefficient D20, w of 6.56 F, resulting in a molecular weight of 115000 for the catalytically active form . Thus, the enzyme is active as the tetramer . So far the enzyme from P . fluorescens is the sole 6-phospho-D-gluconate dehydrogenase (decarboxylating) composed of four polypeptide chains. J Dairy Res, 1983 Feb, 50(1), 9 - 16 Evaluation of the direct epifluorescent filter technique for assessing the hygienic condition of milking equipment; Hunter AC et al.; The hygienic condition of 6 milking installations, 3 sanitized by circulation cleaning (CC) with chlorine-based chemicals and 3 by flushing with acidified boiling water (ABW), was tested using rinses of quarter strength Ringer's solution . The bacterial content of the rinses was determined using both colony counts and the direct epifluorescent filter technique (DEFT) . A comparison of testing methods gave correlation coefficients between colony count and DEFT of 0.82 for plants using CC and 0.46 for plants using ABW . Five strains of bacteria belonging to different genera and commonly found on milking equipment were exposed to various degrees of heat and to various concentrations of chlorine . The effects of such treatments on the staining characteristics of the organisms were studied . It was observed that Staphylococcus aureus and Streptococcus lactis, although killed by heat treatment, stained a bright orange when treated with acridine orange dye . Pseudomonas fluorescens, Escherichia coli and vegetative cells of Bacillus cereus did not take up the orange stain after heat treatment, nor did any of the 5 strains stain orange after treatment with NaOCl . It is suggested that the DEFT is a useful and rapid means of counting bacteria in rinses of equipment where sterilization is due primarily to chlorination, but in the absence of a stain which can differentiate more accurately between dead and living organisms its use is limited where sterilization is carried out solely by heat. Appl Environ Microbiol, 1983 Jan, 45(1), 218 - 22 Properties of alkyl hydroxycinnamates and effects on Pseudomonas fluorescens; Baranowski JD et al.; Alkyl esters of six hydroxycinnamic acids, shown to be active antimicrobial agents when tested against Pseudomonas fluorescens, were further investigated for their effects against this organism . There was no statistically significant adaptation by this organism to either of the methyl esters of caffeic, rho-coumaric, cinnamic, or rho-hydroxybenzoic acids . Mixtures of these compounds taken two at a time gave at least additive effects, with some mixtures showing synergism . Preliminary work was also performed to determine the mode of inhibitory action for these compounds . The inhibition of oxygen utilization by the methyl esters correlated well with growth inhibition . Short-term lethality studies showed that none of the alkyl esters or methyl or propyl paraben produced any bacteriocidal effects . Oil-water partition coefficients were determined for these compounds and were shown to have no correlation with growth inhibitions . These all point to a specific mode of action, based in part on cellular energy depletion, rather than the nonspecific membrane-disrupting effects of other phenolic antimicrobial agents. Biochemistry, 1982 Dec 21, 21(26), 6639 - 46 A study of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens: chemical modification of histidine residues; Wijnands RA et al.; The flavoprotein p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens is inactivated by diethyl pyrocarbonate . Below pH 7, diethyl pyrocarbonate reacts specifically with histidine residues . The inactivation reaction is biphasic and follows pseudo-first-order kinetics . Four of the nine histidine residues of the enzyme are modified . During the first phase of the reaction, one histidine residue is modified and leads to a loss of about 30% of the activity . Modification of the additional three histidine residues during the second phase leads to complete loss of activity . Two of the latter histidine residues are essential for activity and are involved in the binding of reduced nicotinamide adenine dinucleotide phosphate (NADPH) . The activity can be restored almost quantitatively upon treatment of modified enzyme with hydroxylamine . The modified enzyme is still capable of binding NADPH . The dissociation constant of the enzyme-NADPH complex is larger by a factor of 10 for the modified enzyme as compared to that for the native enzyme . The modification does not affect the affinity of the enzyme for the substrate, although effectors protect two histidine residues from chemical modification by diethyl pyrocarbonate . The rate of inactivation of the enzyme is pH dependent and increases with increasing pH values . From the pH dependence of the rate constant, it is calculated that two cooperative histidine residues participate in the reaction with diethyl pyrocarbonate . Both histidine residues possess a pKa' value of 6.2 . At pH greater than 7, other reactions take place which are completely abolished in the presence of an effector (substrate) of the enzyme. Antimicrob Agents Chemother, 1982 Dec, 22(6), 1051 - 7 Lysis of Trypanosoma cruzi by Pseudomonas fluorescens; Mercado TI et al.; Trypomastigotes of Trypanosoma cruzi isolated from the blood of infected mice were lysed within 24 h by an extracellular substance produced by Pseudomonas fluorescens . Isolation of the anti-trypanosomal factor (ATF) was accomplished by growth of the organisms in a defined medium, extracellular secretion by the sedimented cells, sterilization by filtration, lyophilization, dialysis, and gel filtration . Chromatographic separation with Sephadex G-25 and G-200 disclosed the occurrence of three active fractions . ATF-I(1) exhibited a molecular weight higher than 440,000 . ATF-II and ATF-III were considerably smaller (molecular weights approximately 1,355 and 1,060, respectively) . The lytic substance contained protein and lipopolysaccharide, was resistant to heat and freezing, was not proteolytic or hemolytic, and was not inhibited by trypsin but was suppressed by pronase. Appl Environ Microbiol, 1982 Sep, 44(3), 521 - 4 Detection of glucose oxidation products in chilled fresh beef undergoing spoilage; Farber JM et al.; The fate of nutrients during storage of longissimus dorsi muscle at 4 degrees C was examined . Glucose concentrations in meat were shown to decrease concomitantly with an approximately fourfold increase in the activity of glucose dehydrogenase . Gluconate concentrations in meat were determined by an enzyme assay and shown to increase from 2.1 to 40.6 microgram/g upon storage of the meat from day 0 to day 6 . At day 12, gluconate concentrations had decreased to 5.8 microgram/g . Dark firm dry meat, which contains little or no glucose, did not exhibit the same rise and fall in gluconate concentration . Thin-layer chromatographic analysis confirmed the presence of 2-ketogluconate in 6- and 12-day-old longissimus dorsi muscle that had been stored at 4 degrees C . Gluconate concentrations in irradiated sterile meat inoculated with Pseudomonas fluorescens increased from 4.2 to 77.8 microgram/g during the first 6 days of storage at 4 degrees C . Therefore, glucose in meat stored at 4 degrees C appeared to be converted to gluconate, 2-ketogluconate, or both extracellularly by one of the main meat spoilage organisms, most likely the pseudomonads. Biull Eksp Biol Med, 1982 Sep, 94(9), 41 - 2 {Photosensitized oxidation of asparagine-glutamine deamidase from Pseudomonas fluorescens}; Kozlov EA et al.; The process of oxidation of deamidase of asparagine-glutamine from Pseudomonas fluorescens sensitized with dyes has been studied . Oxidation brings about complete inactivation of the enzyme which, however, retains the quaternary structure . In the course of photooxidation, tryptophan residues are first destroyed and then those of histidine . Possibilities of the participation of these amino acid residues in catalysis and maintenance of active conformation of the enzyme are discussed. Biochim Biophys Acta, 1982 Aug 6, 717(2), 376 - 83 Isolation and general characterization of a heat-stable proteinase from Pseudomonas fluorescens aft 36; Stepaniak L et al.; An extracellular proteinase from Pseudomonas fluorescens, strain AFT 36, was isolated to homogeneity by chromatography on DEAE-cellulose and Sephadex G-150; a 230-fold increase in specific activity with a recovery of 53% was obtained . The enzyme was optimally active at pH 6.5 and 45 degrees C; activity declined rapidly at higher temperatures but significant activity persisted down to 4 degrees C . Activity was strongly inhibited by 10(-3) M EDTA and was partially restored by addition of Zn2+, Ca2+ or Co2+ . The Km values on methylated casein and sodium caseinate were 18.2 and 7.1 mg/ml, respectively . The enzyme was very labile in phosphate buffer and in a mild salts buffer at 55 degrees C but was very stable in the latter at more than 80 degrees C. Can J Microbiol, 1982 Aug, 28(8), 907 - 15 The role of some membrane dehydrogenases in Pseudomonas fluorescens; Lynch WH; The membrane-associated oxidative enzymes glucose, gluconate, and malate dehydrogenases were examined in psychrotrophic Pseudomonas fluorescens . The function and activity of these enzymes was determined by measuring extracellular product formation by washed cell suspensions . Membrane dehydrogenase activities and corresponding transport activities for the substrates glucose, gluconate and malate were compared in batch cultures grown with these substrates at 30 and 5 degrees C . These activities correlated with the production or lack of extracellular oxidation products . in chemostat cultures grown at 30 and 5 degrees C, the membrane enzymes and production of their extracellular oxidation products appeared to be regulated by the available carbon concentration . The enzymes were induced and high concentrations of extracellular oxidation products were produced under conditions of nitrogen limitation (carbon excess) but not carbon limitation . The lower affinities of the three membrane enzymes for their respective substrates, when compared with the transport systems utilizing the same substrates, correlated with the observed major function of these enzymes in carbon-excess environments . The primary role of the membrane-associated oxidative enzymes, in carbon dissimilation by this psychrotrophic microorganism at low temperatures in carbon-excess environments, was strongly implicated. J Biol Chem, 1982 Jul 25, 257(14), 8086 - 90 Siderochromes from Pseudomonas fluorescens . II . Structural homology as revealed by NMR spectroscopy; Philson SB et al.; Ferribactin and the pyoverdines, siderochromes that are obtained from liquid cultures of Pseudomonas fluorescens cells, have been studied and compared by 1H and 13C NMR spectroscopy . The proton spectra of the iron-free compounds show that the pyoverdines share with ferribactin a common feature, formyl hydroxamic acid groups, that previously had only been observed in hadacidin, and antitumor antibiotic produced by Penicillium frequentans . The 1H and 13C NMR data confirm that ferribactin is a nonapeptide that contains two residues each of lysine and N6-formyl-N6-hydroxyornithine . This corrects an earlier report (Maurer, B., Muller, A., Keller-Schierlein, W., and Zahner, H . (1968) Arch . Mikrobiol . 60, 326-339) ascribing two acetyl hydroxamic acid groups and three lysyl residues to ferribactin . Similarly, the spectroscopic data show that pyoverdine lacks the Glx and Tyr residues present in ferribactin . On the basis of the compositional analogy exhibited by pyoverdine and ferribactin, it is suggested that the two siderochromes may be metabolically related . The 13C NMR spectra of pyoverdine indicate that its fluorescent component is a nine-carbon aromatic heterocycle, probably identical with an o-dihydroxyquinoline chromophore found in pseudobactin, a fluorescent siderophore produced by Pseudomonas B10. J Biol Chem, 1982 Jul 25, 257(14), 8081 - 5 Siderochromes from Pseudomonas fluorescens . I . Isolation and characterization; Philson SB et al.; Several iron-binding pigments (siderochromes) produced by Pseudomonas fluorescens have been isolated and partially characterized . They include ferribactin and various forms of pyoverdine, as well as some previously unreported compounds . In particular, the existence of ferribactin has been independently confirmed for the first time . Column and thin layer chromatographic procedures have been developed to fractionate, purify, and identify the siderochromes . We find ferribactin to contain nine amino acids, one residue each of glutamine, tyrosine, and glycine, and two each of serine, lysine, and N-hydroxyornithine, rather than 10 as earlier reported . Pyoverdine is a peptide with the same composition as ferribactin except for the absence of glutamine and the substitution of a fluorescent chromophore for tyrosine . Paper electrophoresis reveals an extra ionizable group in ferric pyoverdine relative to pyoverdine or ferribactin which provides that complex a definite cathodic mobility at pH 3 . Optical spectra of the pyoverdine fluorescent component indicate that, in conjunction with the two hydroxamate groups, it is involved in the metal ion coordination, conferring on pyoverdine a dramatically increased affinity for Fe(III) relative to ferribactin. Infect Immun, 1982 Jul, 37(1), 166 - 71 Monoclonal antibodies against Pseudomonas aeruginosa outer membrane antigens: isolation and characterization; Hancock RE et al.; Hybridomas secreting monoclonal antibodies specific for Pseudomonas aeruginosa outer membrane antigens were isolated . One of the antibodies was highly specific for the O antigen of the lipopolysaccharide of International Antigen Typing Scheme serotype 5 strains, reacting only weakly with a serotype 17 strain and failing to react with the outer membranes of strains representing 15 other serotypes . This monoclonal antibody was able to agglutinate heat-killed bacterial cells as well as lipopolysaccharide-coated sheep erythrocytes . Two other monoclonal antibodies were able to interact with the outer membranes of strains representing all 17 serotypes, although they were unable to agglutinate heat-killed bacterial cells . One of these was shown to be specific for the major outer membrane lipoprotein H2 . The antigenic site against which this monoclonal antibody reacted was present in the outer membranes of two Pseudomonas fluorescens strains, two Pseudomonas putida strains, a Pseudomonas anguilliseptica strain, and an Azotobacter vinelandii strain, but not in the outer membranes of five other bacterial species. Biochim Biophys Acta, 1982 Jun 4, 704(2), 385 - 8 Primary structure of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens; Weijer WJ et al.; The amino acid sequence of the p-hydroxybenzoate hydroxylase (4-hydroxybenzoate,NADPH:oxygen oxidoreductase (3-hydroxylating), EC 1.14.13.2) monomer from Pseudomonas fluorescens has been determined . The sequence was elucidated by a combination of the results from an X-ray crystallographic study at 0.25 nm resolution (Wierenga, R.K., de Jong, R.J., Kalk, K.H., Hol, W.G.J . and Drenth, J . (1979) J . Mol . Biol . 131, 55-73) and from protein sequence analysis . The polypeptide chain of the monomer contains 394 amino acids and has a molecular weight of 44 299. J Biochem (Tokyo), 1982 May, 91(5), 1555 - 61 Hydrophobic-ionic chromatography: its application to microbial glucose oxidase, hyaluronidase, cholesterol oxidase, and cholesterol esterase; Sasaki I et al.; Glucose oxidase from Aspergillus niger, hyaluronidase from Streptomyces hyalurolyticus, and cholesterol oxidase and cholesterol esterase from Pseudomonas fluorescens were effectively adsorbed on an Amberlite CG-50 column, when the cell-free cultured medium or the cultured medium with cell extract and without cell debris was applied without desalting but at pH less than or equal to 4.5 . At the acidic pH, all the ion-exchange groups (-COOH) exist in the protonated form; the adsorption is not due to electrostatic attraction, but to hydrophobic interaction . The enzymes thus adsorbed were effectively eluted by increasing pH, at which the ion-exchange groups became dissociated . This type of adsorption-elution is called hydrophobic-ionic chromatography . By a single run of chromatography, glucose oxidase, hyaluronidase, cholesterol oxidase, and cholesterol esterase were purified 30-fold, 12-fold, 45-fold, and 20-fold with yields of 82%, 83%, 80%, and 90%, respectively . This indicates that hydrophobic-ionic chromatography on an Amberlite CG-50 column is effective for the purification of various enzymes, provided that they are stable at the acidic pH. Eur J Biochem, 1982 Apr, 123(3), 547 - 52 D-Malic enzyme of Pseudomonas fluorescens; Knichel W et al.; By the enrichment culture technique 14 gram-negative bacteria and two yeast strains were isolated that used D(+)-malic acid as sole carbon source . The bacteria were identified as Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas aeruginosa and Klebsiella aerogenes . In cell-free extracts of P . fluorescens and P . putida the presence of malate dehydrogenase, D-malic enzyme (NAD-dependent) and L-malic enzyme (NADP-dependent) was demonstrated . D-Malic enzyme from P . fluorescens was purified . Stabilization of the enzyme by 50 mM ammonium sulphate an 1 mM EDTA was essential . Preparation of D-malic enzyme that gave one band with disc gel electrophoresis showed a specific activity of 4-5 U/mg . D-Malic enzyme requires divalent cations . The Km values were for malate Km = 0.3 mM and for NAD Km = 0.08 mM . The pH optimum for the reaction was found to be in the range of pH 8.1 to pH 8.8 . D-Malic enzyme is partially inhibited by oxaloacetic acid, meso-tartaric acid, D-lactic acid and ATP . Determined by gel filtration and gradient gel electrophoresis, the molecular weight was approximately 175 000. Mikrobiologiia, 1982 Mar-Apr, 51(2), 212 - 5 {Dependence of extracellular proteases synthesis on the growth phase of Pseudomonas fluorescens}; Mikel'saar PCh et al.; The aim of this work was to study the dependence of the synthesis of exocellular proteolytic enzymes on the growth phase of Pseudomonas fluorescens which was cultivated in media containing amino acids as a sole carbon source . In all of the studied cases, the proteolytic activity of the medium increased in parallel to the bacterial growth until the culture reached the stationary growth phase . Chloramphenicol added in the phase of intensive growth ceased the increase of the proteolytic activity of the medium without a noticeable delay in time . This fact indicates that the processes of translation and secretion of exoproteases occur virtually at the same time in the cell of P . fluorescens . The rate of degradation of exocellular proteolytic enzymes was shown to be insignificant in comparison to the rate of their synthesis . Therefore, the increase in the proteolytic activity of the medium directly reflects the rate of synthesis of exoproteases. Arzneimittelforschung, 1982, 32(1), 49 - 55 Pharmacological studies of a new non-steroidal antiinflammatory drug: 2-(5-ethylpyridin-2-yl)benzimidazole (KB-1043); Ito K et al.; 2-(5-Ethylpyridin-2-yl)benzimidazole (KB-1043) is a new benzimidazole derivative with marked antiinflammatory, analgesic and antipyretic properties . KB-1043 possesses potent inhibitory activities on the acute inflammatory edema induced by chemical and physical irritants in the hind paw of rats, but almost ineffective (similar to tiaramide) against adjuvant-induced polyarthritis in the rat . The antagonistic effects of KB-1043 on increased vascular permeability are more potent than those of phenylbutazone, and the analgesic effects of KB-1043 on the pain induced by chemical and mechanical stimulations were nearly equal to, or slightly less potent than, those of tiaramide and phenylbutazone . The fevers induced by brewer's yeast and polysaccharide from Pseudomonas fluorescens (T.T.G.) were lowered by KB-1043 . THe ulcerogenic activity of KB-1043 was weaker than those of tiaramide and phenylbutazone . The therapeutic index of KB-1043 therefore is superior to those of tiaramide and phenylbutazone. Rev Esp Fisiol, 1982, 38 Suppl, 1 - 27 {Topography of the metabolic cycle of 4-aminobutyrate}; Santos-Ruiz A; This work describes, with some detail the intervention of 4-aminobutyrate as protagonist of a derivation of tricarboxylic cycle . Its vicarial mission is emphasized in connection with its existence in microorganisms (Escherichia coli and Pseudomonas fluorescens), plants (Helianthus tuberosus . Lupinus albus and Agave americana), neoplasic cells (ascitic tumor of Ehrlich and HeLa cells) and animal tissues (adrenal medulla and brain. Arch Environ Contam Toxicol, 1982, 11(2), 203 - 7 Effect of sequence of exposure to chlorophenols in short-term bacterial bioassays; Trevors JT et al.; A bioassay using Pseudomonas fluorescens was affected by the sequence of exposure to pentachlorophenol (PCP) and 2,3,4,5-tetrachlorophenol (TCP) . Surviving cells from standardized cell suspensions initially treated with PCP at concentrations ranging from 10 to 75 micrograms/ml, followed by removal of the toxicant, were not affected by a second dose of PCP at the same concentration . However, if the second dose was TCP, the test organism was sensitive to the second exposure . The most toxic sequence was an initial exposure to TCP followed by a second exposure to PCP . The response of the test organism to PCP and TCP was clearly dependent upon both the toxicant concentrations used and the sequence of toxicant addition. C R Seances Soc Biol Fil, 1982, 176(5), 700 - 6 {Influence of diverse nucleotide constituents of Pseudomonas fluorescens on the D-glucose-dehydrogenase activity of the bacterium}; Wurtz B; The D-glucose-dehydrogenase activity of P . fluorescens is enhanced or depressed by some nucleotides synthetised by this bacterium: stimulatory nucleotides have been separated from inhibitory . UTP exhibits a non-competitive inhibition upon the pure soluble enzyme. Curr Eye Res, 1982-83, 2(5), 289 - 93 Adherence properties of Pseudomonas pili to epithelial cells of the human cornea; Reichert RW et al.; In vitro adherence of Pseudomonas fluorescens organisms to the human cornea is correlated with bacterial pili . The role of pili in the attachment to human corneal epithelial cells was studied in an in vitro adherence assay system . A homogeneous, purified pilin preparation showed a molecular weight of 16,500 on SDS-polyacrylamide gels . Within 5 minutes incubation, 5.5 pg of pilin adhere to 15,000 epithelial cells . When epithelial cells were preincubated with purified pilin, a subsequent decrease in adherence of labeled pilin was noted . It appears that pili mediate adherence of Pseudomonas organism to human cornea. Gene, 1981 Dec, 16(1-3), 149 - 59 Expression of Pseudomonas fluorescens D-galactose dehydrogenase in E . coli; Buckel P et al.; To investigate the heterologous expression of Pseudomonas genes in Escherichia coli we have cloned P . fluorescens DNA in an E . coli {cosmid} system . A colony bank representing the whole P . fluorescens chromosome was screened immunologically using a modification of the method described by Broome and Gilbert (1978) . Radioactive labelling of the antibodies was replaced by conjugation with horseradish peroxidase . Among 523 E . coli colonies one was D-galactose dehydrogenase-positive . The expression of this enzyme in primary clones was lower than in the uninduced Pseudomonas . Subcloning of the D-galactose dehydrogenase gene, in vitro mutagenesis of the DNA, and coupling to a strong E . coli promoter yielded an E . coli strain that produces 90 times more of the enzyme than the induced P . fluorescens. J Gen Microbiol, 1981 Jul, 125(Pt 1), 217 - 20 Isolation of alginate-producing mutants of Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas mendocina; Govan JR et al.; Spontaneous alginate-producing (muc) variants were isolated from strains of Pseudomonas fluorescens, P . putida and P . mendocina at a frequency of 1 in 10(8) by selecting for carbenicillin resistance . The infrared spectrum of the bacterial exopolysaccharide was typical of an acetylated alginate similar to that previously described in Azotobacter vinelandii and in mucoid variants of P . aeruginosa . Mucoid variants were not isolated from P . stutzeri, P . pseudoalcaligenes, P . testosteroni, P . diminuta, P . acidovorans, P . cepacia or P . maltophilia. J Dairy Sci, 1981 Jul, 64(7), 1545 - 50 Unique response to heat of extracellular protease of Pseudomonas fluorescens M5; Marshall RT et al.; Eight proteases selected from 38 cultures of psychrotrophic bacteria were subjected to heat stability tests . Cell-free filtrates of the broth in which the cultures were grown individually were heated to 40, 50, 60, and 70 degree C for 0, 15, 30, and 60 min . Six of the filtrates were less than 50% inactivated by heating at 40 degree C for 60 min, whereas the enzyme of Pseudomonas fluorescens M5 was inactivated completely by this treatment . Of the five most proteolytic cultures tested, including P . fluorescens M5, losses in protease activity ranged from 9 to 34% on heating at 70 degree C for 60 min . Purified M5 protease retained at least 75% of its activity over the pH range of 4.5 to 7.5 . Electrophoresis of active M5 protease from four chromatographed fractions revealed two bands in each with approximate molecular weights of 35,000 and 45,000 . Heating at 40 degree C did not alter mobility of either band . Reasons for lability at 40 degree C but stability at 50, 60, and 70 degree C are discussed . Complexation with casein, observed with another protease, was not a possible explanation for stability at 40 degree C. Biochimie, 1981 Jun, 63(6), 535 - 40 Inducible catalase in Pseudomonas fluorescens; Rodriguez-Bravo S et al.; The catalase activity of a non-proliferating suspension of Pseudomonas fluorescens doubled after six hours incubation in a 50 mM phosphate buffer medium (pH 7.3) . The same effect was observed in a peptone medium . The increased activity was due to induced enzyme synthesis, and not to activation of preexisting catalase . Induced catalase was separated by electrophoresis from deuterium labelled constitutive catalase . The enzyme was also induced under anaerobic conditions in phosphate buffer or in culture when nitrate was supplied as an electron acceptor . Induction was considerably increased by the addition of various nucleotides and amino acids to the incubation medium. J Bacteriol, 1981 Jun, 146(3), 1003 - 12 Mobilization of the genes for photosynthesis from Rhodopseudomonas capsulata by a promiscuous plasmid; Marrs B; Plasmid pBLM2, a derivative of RP1 with enhanced chromosome mobilization activity in Rhodopseudomonas capsulata, was isolated by screening rare exconjugant clones for sex factor activity . pBLM2 mobilized all known genes affecting photosynthesis as well as chromosomal genes for streptomycin and rifampin resistance and tryptophan and cytochrome biosynthesis . Tight linkage was exhibited among the genes affecting photosynthesis . The frequency of successful transfer of chromosomal markers reached 6 X 10(-4) per donor cell . R-primes were occasionally formed during conjugation, and a number of R-primes bearing the genes for photosynthesis were isolated by screening R . capsulata exconjugants with complementation phenotypes for the ability to transmit plasmid-borne R . capsulata genes to Escherichia coli cells . These R-primes were unstable in R . capsulata, but stable in E . coli or Pseudomonas fluorescens . Complementation and recombination events that occurred upon introduction of R-primes into R . capsulata mutants with altered photosynthetic apparatuses could be visualized as variations in colony pigmentation . Each R-prime studied complemented all known types of mutation affecting the differentiation of the photosynthetic apparatus, and no other R . capsulata gene was identified on those plasmids . The R . capsulata genes borne on the R-primes were not functional in E . coli or P . fluorescens. J Bacteriol, 1981 May, 146(2), 705 - 12 Glutamine synthetase of pseudomonads: some biochemical and physicochemical properties; Meyer JM et al.; The glutamine synthetases from several Pseudomonas species were purified to homogeneity, and their properties were compared with those reported for the enzymes from Escherichia coli and other gram-negative bacteria . The glutamine synthetase from Pseudomonas fluorescens was unique because it was nearly precipitated quantitatively as a homogeneous protein during dialysis of partially purified preparations against buffer containing 10 mM imidazole (pH 7.0) and 10 mM MnCl2 . The glutamine synthetases from Pseudomonas putida and Pseudomonas aeruginosa were purified by affinity chromatography on Affi-blue gel . Dodecamerous forms of the E . coli and P . fluorescens glutamine synthetases had identical mobilities during polyacrylamide gel electrophoresis . Their dissociated subunits, however, migrated differently and were readily separated by electrophoresis on polyacrylamide gels containing 0.1% sodium dodecyl sulfate . This difference in subunit mobilities is not related to the state of adenylylation . Regulation of the Pseudomonas glutamine synthetase activity is mediated by an adenylylation-deadenylylation cyclic cascade system . A sensitive procedure was developed for measuring the average number of adenylylated subunits per enzyme molecule for the glutamine synthetase from P . fluorescens . This method takes advantage of the large differences in transferase activity of the adenylylated and unadenylylated subunits at pH 6.0 and of the fact that the activities of both kinds of subunits are the same at pH 8.45. Arch Microbiol, 1981 Apr, 129(2), 135 - 40 Assimilatory nitrate uptake in Pseudomonas fluorescens studied using nitrogen-13; Betlach MR et al.; The mechanism of nitrate uptake for assimilation in procaryotes is not known . We used the radioactive isotope, 13N as NO3-, to study this process in a prevalent soil bacterium, Pseudomonas fluorescens . Cultures grown on ammonium sulfate or ammonium nitrate failed to take up labeled nitrate, indicating ammonium repressed synthesis of the assimilatory enzymes . Cultures grown on nitrite or under ammonium limitation had measurable nitrate reductase activity, indicating that the assimilatory enzymes need not be induced by nitrate . In cultures with an active nitrate reductase, the form of 13N internally was ammonium and amino acids; the amino acid labeling pattern indicated that 13NO3- was assimilated via glutamine synthetase and glutamate synthase . Cultures grown on tungstate to inactivate the reductase concentrated NO3- at least sixfold . Chlorate had no effect on nitrate transport or assimilation, nor on reduction in cell-free extracts . Ammonium inhibited nitrate uptake in cells with and without active nitrate reductases, but had no effect on cell-free nitrate reduction, indicating the site of inhibition was nitrate transport into the cytoplasm . Nitrate assimilation in cells grown on nitrate and nitrate uptake into cells grown with tungstate on nitrite both followed Michaelis-Menten kinetics with similar Km values, 7 muM . Both azide and cyanide inhibited nitrate assimilation . Our findings suggest that Pseudomonas fluorescens can take up nitrate via active transport and that nitrate assimilation is both inhibited and repressed by ammonium. Biochemistry, 1981 Mar 17, 20(6), 1481 - 90 Mechanism of action of glutaryl-CoA and butyryl-CoA dehydrogenases . Purification of glutaryl-CoA dehydrogenase; Gomes B et al.; Glutaryl-CoA dehydrogenase, a flavoprotein, catalyzes the reaction -OOCCH3CH2--CH2COSR (FAD leads to FADH2) leads to CH3CH = CHCOSR + CO2 (SR = CoA or pantetheine) . With the isolated enzyme, a dye serves as the final electron acceptor . The enzyme from Pseudomonas fluorescens (ATCC 11250) has been purified to homogeneity . It was established with appropriate isotopic substitutions that the proton which is added to the gamma position of the product, subsequent to decarboxylation, is not derived from the solvent but is derived from the alpha position of the substrate . Under conditions where no net conversion of substrate occurs, i.e., in the absence of electron acceptor, the enzyme catalyzes the exchange of the beta hydrogen of the substrate with solvent protons . Butyryl-CoA dehydrogenase (M . elsedenii), which catalyzes an analogous reaction, catalyzes the exchange of both the alpha and beta hydrogens with solvent protons in the absence of electron acceptor . Glutaryl-CoA dehydrogenase and butyryl-CoA dehydrogenase are irreversibly inactivated by the substrate analogues 3-butynoylpantetheine and 3-pentynoylpantetheine . These inactivators do not form an adduct with the flavin and probably react with a nucleophile at the active site . Upon inactivation, the spectrum of the enzyme-bound flavin is essentially unchanged, and the flavin can be reduced by Na2S2O4 . We suggest that inactivation involves intermediate allene formation . We proposed that these results support an oxidation mechanism for glutaryl-CoA dehydrogenase and butyryl-CoA dehydrogenase which is initiated by proton abstraction . With glutaryl-CoA dehydrogenase, the base, which abstracts the substrate alpha proton, is shielded from the solvent and is then used to protonate the carbanion (CH2--CH--CHCOSCoA) formed after oxidation and decarboxylation. J Dairy Res, 1981 Feb, 48(1), 115 - 22 Effects of adding potassium iodate to milk before UHT treatment . ii . iodate-induced proteolysis during subsequent aseptic storage; Skudder PJ; The addition of potassium iodate to milk at 9.1 mM before UHT treatment resulted in rapid breakdown of alpha s- and beta-casein during subsequent aseptic storage . Maximum rates of proteolysis were observed at storage temperatures of 37-45 degrees C, but the reaction was strongly inhibited by storage at 55 degrees C and by increased holding time at 140 degrees C during the UHT sterilization . Iodate-induced proteolysis of purified alpha s1-and beta-casein was detected only with solutions in the serum phase of raw milk; no proteolysis occurred with solutions in 0.1 M-phosphate buffer (pH 6.7) or in milk ultrafiltrate, irrespective of whether whey proteins and lactose were also added . Thus, it appears that iodate increased the activity of one or more proteolytic components which were present in milk and were unable to pass through an ultrafiltration membrane . However, it is unlikely that iodate acts by increasing the activity of proteinases produced by contaminant bacteria; the presence of iodate did not affect the activity of a proteolytic enzyme isolated from Pseudomonas fluorescens PM-1 . Furthermore, iodate promoted protein breakdown during storage of milk drawn aseptically from the cow and subsequently UHT processed . It is suggested that iodate increased the activity of native milk proteinases, other than plasmin which was inactivated by UHT treatment, possibly by preventing thiol-disulphide exchange reactions during the heating process. Am J Clin Pathol, 1981 Feb, 75(2), 211 - 3 Inability of the API-20E system to speciate the fluorescent group of pseudomonads; Woolfrey BF et al.; One thousand randomly selected clinical isolates of the fluorescent group of Pseudomonas were speciated in parallel by the API-20E system and by a classic microbiologic 17-test battery . The classic battery identified the isolates as 993 Pseudomonas aeruginosa, five Pseudomonas putida, and two Pseudomonas fluorescens . To augment the P . putida and P . fluorescens data, 52 reference isolates were also tested in parallel . API-20E was found to identify 99% of P . aeruginosa as P . aeruginosa . However, the 59 P . putida and P . fluorescens isolates were not accurately identified and were designated 58% P . aeruginosa, 24% P . stutzeri, 16% ambiguous categories, and only 2% in agreement with the classic designation . These findings indicated that the API-20E system does not accurately identify the non-P . aeruginosa fluorescent pseudomonads and that errors in P . aeruginosa designations will increase as the percentages of P . putida and P . fluorescens increase in the population sample. Mol Cell Biochem, 1981 Jan 28, 34(2), 95 - 9 Acetylcholinesterase from rat red cells and cholinesterase of Pseudomonas aeruginosa: different types of inhibition by atropine; Domenech CE et al.; The inhibition by atropine of cholinesterase from Pseudomonas aeruginosa has been studied in parallel with the membrane bound acetylcholinesterase from rat red cells . Acetylcholinesterase of rat red cells, like other animal cholinesterases, was competitively inhibited while the cholinesterase from Pseudomonas aeruginosa was partially non competitively inhibited by atropine . These results clearly indicated a different behavior of cholinesterase from Pseudomonas aeruginosa in comparison with the enzyme of Pseudomonas fluorescens and other animal cholinesterases. Eur J Biochem, 1980 Dec, 113(1), 151 - 7 The amino-acid sequence of the three smallest CNBr peptides from p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens; Vereijken JM et al.; After CNBr cleavage of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens, five peptides and free homoserine were isolated (see preceding paper in this journal) . The amino acid sequences of the three smallest peptides, viz . CB3, CB4 and CB5, were determined by automated Edman degradation and analysis of enzymatic subdigests . These peptides form a continuous stretch of 110 residues from the N terminus: (Formula: See Text). Eur J Biochem, 1980 Dec, 113(1), 141 - 50 Primary and tertiary structure studies of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens . Isolation and alignment of the CNBr peptides; interactions of the protein with flavin adenine dinucleotide; Hofsteenge J et al.; p-Hydroxybenzoate hydroxylase from Pseudomonas fluorescens contains six methionine residues, one of which is N-terminal . After CNBr cleavage five peptides, ranging from 13 to 158 residues in length, and free homoserine were isolated and purified by repeated gel filtration . The alignment of the CNBr fragments was deduced from a 0.25-nm electron density map and sequence data . The isolated fragments account for the entire polypeptide chain . The amino acid sequence of the N-terminal quarter of the polypeptide chain was determined . The X-ray results together with the sequence data yielded details of the binding of FAD . The AMP moiety was bound to a beta alpha beta unit resembling that found in the dehydrogenases . Hydrogen bonds were present between the protein and the ribityl residue and the isoalloxazine ring . Furthermore, a homology was found between the N-terminal amino acid sequence of p-hydroxybenzoate hydroxylase and another enzyme containing FAD, viz . D-amino acid oxidase . This finding suggests the presence of a mononucleotide binding fold at the N terminus of the latter. Mikrobiologiia, 1980 Nov-Dec, 49(6), 888 - 92 {Resting state in Pseudomonas fluorescens induced by a prolonged water deficit}; Shevtsova II et al.; Various strains of Pseudomonas fluorescens showed different resistance to water deficiency in soils with a low water content (alpha w = 0.75) . Humidification of dry soil reactivated the cultures . The period of reactivation took from several hours to three days and depended on the nature of inoculated strains . The growth of various strains of Pseudomonas fluorescens was studied in the conditions of a low water content in soil; the cells were investigated by electron microscopy before and after soil humidification . The results suggest that this organism can be in a certain "resting state" which is characterized by changes in the cell structure and by the time it takes the cells to reactivate after humidification of a dry soil sample. J Gen Microbiol, 1980 Oct, 120(Pt 2), 485 - 511 Evolution in Pseudomonas fluorescens; Champion AB et al.; The relationships among 93 strains of Pseudomonas fluorescens were investigated by (1) a numerical taxonomic analysis on the results of 150 phenotypic tests, (2) DNA hybridization studies using 16 reference strains, (3) quantitative microcomplement fixation studies using six reference strains with antibodies directed against the protein azurin . In general, the strains fell into distinct clusters . Assignment to these clusters on the basis of azurin immunological similarity showed 98% agreement with assignment based on DNA homology, suggesting that many genes will follow the same pattern . Of the strains that clustered on the basis of genotype (DNA, azurin) 88% also clustered on the basis of phenotype . The occasional noncongruency observed between the genotypic and phenotypic data may be due to the variable rates of phenotypic evolution . These results provide a perspective on the roles of horizontal and vertical transfer of genes in the evolution of this bacterial group. J Clin Microbiol, 1980 Oct, 12(4), 521 - 6 Headspace analysis of volatile metabolites of Pseudomonas aeruginosa and related species by gas chromatography-mass spectrometry; Labows JN et al.; Gas chromatographic-mass spectrometric analysis of headspace volatiles was performed on cultures of 11 strains of Pseudomonas aeruginosa and 1 strain each of Pseudomonas cepacia, Pseudomonas putida, Pseudomonas putrefaciens, Pseudomonas fluorescens, and Pseudomonas maltophilia . All strains of Pseudomonas aeruginosa produced a distinctive series of odd-carbon methyl ketones, particularly 2-nonanone and 2-undecanone, and 2-aminoacetophenone . The other strains failed to produce 2-aminoacetophenone . Two sulfur compounds, dimethyldisulfide and dimethyltrisulfide, were present in strains of P . aeruginosa and in variable amounts in other species . Butanol, 2-butanone, 1-undecene, and isopentanol were also detected in P . aeruginosa cultures. Biokhimiia, 1980 Oct, 45(10), 1850 - 8 {Structural organization of the bacterial nucleoid using endonucleolysis}; Kharchenko EP et al.; The fragmentation of bacterial deoxyribonucleoprotein (bDNP) in the spheroplasts of Escherichia coli, Serratia marcescens, Pseudomonas fluorescens and Micrococcus luteus by bacterial intracellular Ca2+ or Ca2+, Mg2+-dependent endonucleases in situ was studied . An electrophoresis of the extracted nuclease-split bDNP revealed the presence of high molecular weight (nuclease-resistant) and low molecular weight multiple fragments (100--120 nucleotide pairs) . The electrophoretic mobility of the smallest nuclease-split DNA fragments in all bacterial species under study was similar, indicating the orderly structure of bDNP . Two total fractions whose electrophoretic mobility corresponded to that of the histones H2a and H2b from calf thymus were prevalent in the spectrum of acid-soluble bDNP proteins of gram-negative species . The heterogeneity of DNP with respect to its sensitivity to nucleases, is interaction with membranes and protein distribution pattern were revealed by treatment of the bacterial nucleoid with endogenous endonucleases, which probably reflects differences in the functional state of individual sites of the genome. Mikrobiologiia, 1980 Sep-Oct, 49(5), 708 - 14 {Localization of metabolites which are regulators of extracellular protease synthesis in Pseudomonas fluorescens bacteria}; Vilu RO et al.; The effect of over forty low molecular weight substrates on the growth and synthesis of exocellular neutral proteases was studied in Pseudomonas fluorescens . Neutral exoproteases were found to be regulated enzymes . Glucose did not repress the synthesis of exocellular proteases; the regulation of their synthesis by amino acids involved the mechanism of induction . The data suggest that the primary intracellular inductors of the synthesis of exoproteases are formed for different groups of amino acids at different levels of their utilization by the cells, viz . at the level of transport and at the level of the first steps in the degradation of their carbon backbones . The paper discusses possible molecular mechanisms for integrating the signal of the primary intracellular inductors, which directly regulate the activity of the operon(s) of neutral exocellular proteases. Biochim Biophys Acta, 1980 Aug 7, 614(2), 266 - 73 Evidence for the importance of cysteine and arginine residues in Pseudomonas fluorescens UK-1 pantoate dehydrogenase; Myohanen T et al.; Homogeneous pantoate dehydrogenase (D-pantoate:NAD+ 4-oxidoreductase, EC 1.1.1.106) was shown to be sensitive to inactivation by p-chloromercuribenzoate (100 microM), 5,5'-dithiobis(2-nitrobenzoic acid) (1 mM), iodoacetic acid (1 mM) and phenylglyoxal (5.3 mM) . Potassium D-pantoate and NAD protected against inactivation by p-chloromercuribenzoate, 5,5'-dithiobis (2-nitrobenzoic acid) and iodoacetic acid . NAD and D-pantoate also provided substantial protection against inactivatrion by phenylglyoxal . Titration of the sulphydryl groups by 5,5'-dithiobis(2-nitrobenzoic acid) and incorporation of {14C}carboxymethyl revealed that there are two cysteine residues which are modified and one of those is essential for activity . In the presence of NAD and D-pantoate, incorporation of {14C}phenylglyoxal was decreased by 0.42 mol/mol of subunit. J Antibiot (Tokyo), 1980 Aug, 33(8), 791 - 5 B-1008, a new antibiotic of bacterial origin containing a spermidine moiety . Production, isolation, characterization and biological properties; Kido Y et al.; A new antibiotic, B-1008 has been isolated from the cultured broth of Pseudomonas fluorescens No . 101 B-13L . B-1008 is a water-soluble basic substance containing the spermidine moiety and possessing antibacterial activity against a wide range of bacterial species. J Bacteriol, 1980 Jul, 143(1), 338 - 42 Effect of temperature on Pseudomonas fluorescens chemotaxis; Lynch WH; The effects of temperature and attractants on chemotaxis in psychrotrophic Pseudomonas fluorescens were examined using the Adler capillary assay technique . Several organic acids, amino acids, and uronic acids were shown to be attractants, whereas glucose and its oxidation products, gluconate and 2-ketogluconate, elicited no detectable response . Chemotaxis toward many attractants was dependent on prior growth of the microorganism with these compounds . However, the organic acids, malate and succinate, caused strong chemotactic responses regardless of the carbon source used for growth of the bacteria . The temperature at which the cells were grown (30 or 5 degrees C) had no significant detectable effect on chemotaxis to the above attractants . The temperature at which the cells were assayed appeared to affect the rate but the extent of the chemotactic response, nor the concentration response curves . The ratios of the rate of accumulation of cells to the attractant malate were approximately 2, 4, and 1 at 30, 17, and 5 degrees C, respectively . Strong chemotactic responses were observed with cells assayed at temperatures approaching 0 degree C and appeared to be functional over a broad temperature range of 3 to 35 degrees C. Biochim Biophys Acta, 1980 Jun 13, 613(2), 266 - 74 Comparison of D-malate and beta, beta-dimethylmalate dehydrogenases from Pseudomonas fluorescens UK-1; Lahdesmaki M et al.; D-Malate dehydrogenase (D-malate:NAD+ oxidoreductase (decarboxylating), EC 1.1.1.83) was purified to homogeneity from Pseudomonas fluorescens UK-1 grown on D-malate and some properties of the purified enzyme were compared with those of beta, beta-dimethylmalate dehydrogenase (3,3-dimethyl-D-malate:NAD+ oxidoreductase (decarboxylating), EC 1.1.1.84) . D-Malate dehydrogenase has the molecular weight and subunit size of 140 000 and 34 000, respectively (the same as those of beta, beta-dimethylmalate dehydrogenase) . The amino acid compositions of the two enzymes are similar as well . D-Malate dehydrogenase cross-reacted with anti-beta, beta-dimethylmalate dehydrogenase and beta, beta-dimethylmalate dehydrogenase cross-reacted with anti-D-malate dehydrogenase . The two dehydrogenases have similar catalytical properties . Both of the dehydrogenases were unaffected by sulphydryl reagents but were inactivated by 1,2-butanedione . NAD provided better protection against inactivation than D-malate or beta,beta-dimethyl-DL-malate. Can J Microbiol, 1980 Apr, 26(4), 554 - 5 Identification of Pseudomonas aeruginosa with the API-20E system; Amato S et al.; The API-20E multitest system was found to be capable of identifying Pseudomonas aeruginosa isolates at least to the level of the Pseudomonas fluorescens group within 18 h at 35 degrees C . Additional tests, such as the acetamide reaction, growth at 42 degrees C, and the oxidative (OF) glucose test, assisted in the speciation of P . aeruginosa. Acta Chem Scand B, 1980, 34(6), 423 - 7 D-Malate dehydrogenase from Pseudomonas fluorescens UK-1; Lahdesmaki M et al.; An enzyme catalyzing oxidative decarboxylation of D-malate was induced by D-malate and beta, beta-dimethyl-malate in Pseudomonas fluorescens UK-1 . D-Malate dehydrogenase was purified to homogeneity from the cells grown on D-malate by using ammonium sulfate precipitation, heat treatment, DEAE-sephadex chromatography, Ultrogel AcA 34 gel filtration and 5'-AMP-Sepharose affinity chromatography . D-Malate dehydrogenase has a molecular weight of 140 000 and contains 4 identical subunits of 34 000 . The enzyme is very stable against sulfhydryl group reagents or alkylating agents and rather stable against heat or protein denaturants like urea or sodium dodecyl sulfate . NAD protects against heat inactivation and trypsinization but not against protein denaturants . D-Malate dehydrogenase from Escherichia coli does not cross-react with anti-D-malate dehydrogenase to D-malate dehydrogenase form P.fluorescens UK-1. Folia Microbiol (Praha), 1980, 25(2), 168 - 73 Fluorescent pseudomonads in the rhizosphere of plants and their relation to root exudates; Vancura V; Fluorescent pseudomonads were present in chernozem soil not influenced by plant roots (10(3)-10(4) per g dry soil) in the rhizosphere soil of various plants (10(4)-10(5) per g soil) and on roots (10(3) to 10(7) per g fresh roots), depending on the species and age of the plant . Relative species representation of fluorescent pseudomonads changed on the roots and in the plant rhizosphere as compared with free soil . Pseudomonas fluorescens, representing 60-93% of the population of fluorescent pseudomonads predominated on the roots of all plants investigated . Somewhat different results were obtained in rhizosphere soil . Relatively higher numbers of P . fluorescens were detected in the rhizosphere soil of cucumber and maize, numbers in the rhizosphere soil of wheat were practically the same as in free soil and higher numbers of P . putida were found in the rhizosphere soil of barley . Almost all components contained in the root exudates of the plants studied, including beta-pyrazolylalanine from the root exudates of cucumbers were utilized as carbon and energy sources . Root exudates of wheat and maize were utilized by the strain P . putida K2 with an efficiency of 73-91%, depending on species and age of the plant. Appl Environ Microbiol, 1980 Jan, 39(1), 36 - 40 Microbial lipolysis at low temperatures; Andersson RE; It was found that lipase production during the growth of Pseudomonas fluorescens was not a function of the total number of bacteria . The optimal temperatures for bacterial growth and lipase production were determined as 20 and 8 degrees C, respectively . The lipolytic activity was studied in emulsions of olive oil at temperatures ranging from +8 to -30 degrees C . After an initially rapid lipolysis, the reactions retarded at different levels depending on storage temperature . Transference to a higher temperature resulted in a resumed lipolysis . Also, at low temperatures, lipolysis was studied as a function of water activity and was found to occur in dehydrated substrates. Eur J Biochem, 1979 Nov 1, 101(1), 235 - 44 A study of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens . Improved purification, relative molecular mass, and amino acid composition; Muller F et al.; The purification procedure for p-hydroxybenzoate hydroxylase has been modified by replacement of the DEAE-cellulose (DE-32) column in the original procedure by a Sephadex--Cibacron-blue affinity column . In this way the yield of enzyme could be improved from 16% to about 40--50% . Preparative gel chromatography indicated that the enzyme does not exist as a monomeric species as earlier believed but mainly as a dimer . Sodium dodecyl sulfate gel electrophoresis of purified enzyme revealed a minimum relative molecular mass (Mr) of 43000--45000 . Analytical gel chromatography, sedimentation equilibrium and sedimentation velocity experiments showed that the enzyme exists in solution mainly as a dimer but also in higher-order quaternary structures (presumably tetramer and hexamer) . Temperature dependence of the distribution of the oligomers suggests that the association is of hydrophobic nature . The amino acid composition of the enzyme is also presented . The enzyme contains no disulfide but five sulfhydryl groups . In the native state of the enzyme only one sulfhydryl group is accessible to N-ethylmaleimide or 5,5'-dithiobis(2-nitrobenzoic acid) . The iso-electric point of the enzyme was found to be 5.8. Prikl Biokhim Mikrobiol, 1979 Sep-Oct, 15(5), 671 - 5 {Synthesis of L-aspartic acid by Escherichia coli and Pseudomonas fluorescens as related to the cultivation conditions}; Malofeeva IV et al.; The capacity of the cultures Escherichia coli str . 85, 113, BC, C and K-12 and Pseudomonas fluorescens str . 1 to synthesize L-aspartic acid from fumarate and ammonium ions was studied . E . coli str . 85 was shown to synthesize the largest amounts of aspartic acid . The cultivation conditions which helped to increase the activity several times were selected . The product of fumarate amination by ammonium ions was identified and found to be L-isomer of aspartic acid with an angle of rotation of {alpha} 20/D = +25,5 degrees in 6 N HCl. Appl Environ Microbiol, 1979 Aug, 38(2), 237 - 40 Effect of carbon dioxide on growth of Pseudomonas fluorescens; Gill CO et al.; In minimal medium at 30 degrees C, growth of Pseudomonas fluorescens was stimulated when the pressure (p) of CO2 in solution was 100 mm of Hg, but at higher concentrations the growth rate declined linearly with increasing pCO2 . All concentrations of CO2 were inhibitory for growth in complex medium, and at 30 degrees C the maximum degree of inhibition was attained when pCO2 was 250 mm of Hg . The degree of inhibition at a constant pCO2 in solution increased with decreasing temperature . The degree of inhibition was directly proportional to temperature for growth in complex medium, but not in minimal medium . The inhibition of cell respiration by CO2 was the same whether cells had been grown in air or in the presence of CO2, indicating that adaptive enzyme synthesis does not occur in response to CO2. J Biol Chem, 1979 Jul 25, 254(14), 6657 - 66 Kinetic studies on the reaction of p-hydroxybenzoate hydroxylase . Agreement of steady state and rapid reaction data; Husain M et al.; p-Hydroxybenzoate hydroxylase (EC 1.14.13.2) from Pseudomonas fluorescens is a NADPH-dependent, FAD-containing monooxygenase catalyzing the hydroxylation of p-hydroxybenzoate to form 3,4-dihydroxybenzoate in the presence of NADPH and molecular oxygen . The mechanism of this three-substrate reaction was investigated in detail at pH 6.6, 4 degrees C, by steady state kinetics, stopped flow spectrophotometry, and equilibrium binding experiments . The initial velocity patterns are consistent with a ping-pong type mechanism which involves two ternary complexes between the enzyme and substrates . The first ternary complex is formed by random addition of p-hydroxybenzoate and NADPH to the enzyme, followed by the release of the first product (NADP+) . The reduced enzyme . p-hydroxybenzoate complex now reacts with oxygen, the third substrate, to form the second ternary complex . The enzyme-bound p-hydroxybenzoate then reacts with the activated oxygen to give 3,4-dihydroxybenzoate which is released regenerating the oxidized enzyme for the next cycle . The binding of p-hydroxybenzoate to the oxidized enzyme to form a 1:1 complex causes large, characteristic spectral perturbations and fluorescence quenching . The dissociation constant for the enzyme . substrate complex was obtained by titrations in which absorbance and/or fluorescence quenching was measured . The binding constants of NADPH to the enzyme with and without p-hydroxybenzoate were determined kinetically by measuring the rate of reduction of the enzyme at different concentrations of NADPH . The reduction of the enzyme proceeds extremely slowly in the absence of p-hydroxybenzoate . The presence of the substrate causes a dramatic stimulation (140,000-fold) in the rate of enzyme reduction . The anaerobic reduction of the enzyme by NADPH in the presence of p-hydroxybenzoate produces a transient charge-transfer intermediate . On the basis of the proposed mechanism, the dissociation constants for p-hydroxybenzoate and NADPH as well as the Michaelis constants for all the three substrates were calculated from the initial velocity data . The agreement obtained between various kinetic parameters from the initial rate measurements and those calculated from the individual rate constants determined in rapid reactions, strongly supports the proposed mechanism for the p-hydroxybenzoate hydroxylase reaction. J Biochem (Tokyo), 1979 Jun, 85(6), 1415 - 20 Occurrence of thermolabile and regulatory NAD-linked glutamate dehydrogenase in Pseudomonas fluorescens; Tokushige M et al.; NAD-linked glutamate dehydrogeanse {EC 1.4.1.2} was detected together with NADP-linked glutamate dehydrogenase {EC 1.4.1.4} and aspartase {EC 4.3.1.1} in Pseudomonas fluorescens cells . The three enzymes were distinctly separated by DEAE-Sephadex column chromatography . The NAD-linked enzyme was extremely thermolabile and was rapidly inactivated even at temperatures as low as 35--40 degrees C . The combined addition of NAD+ and glutamate, however, effectively stabilized the enzyme . The glutamate saturation profile of the NAD-linked enzyme exhibited cooperativity with a Hill coefficient (n) of 1.4 . ATP inhibited the enzyme in an allosteric manner, increasing the n value to 2.2 . These results suggest a novel type of metabolic regulation shared by the three enzymes in the biosynthesis and catabolism of amino acids. Appl Environ Microbiol, 1979 Mar, 37(3), 443 - 8 Thermal stress of Pseudomonas fluorescens in complex media; McCoy DR et al.; Pseudomonas fluorescens (P7) cells were stressed by incubation at 43 degrees C for 2 h . The stress induced a 9-h lag in replication after the return of the temperature of the culture to 25 degrees C . Stressed cells demonstrated a sensitivity to diluents and plating media during the recovery period . Data from utilization of selective inhibitors suggested that ribonucleic acid and protein, but not deoxyribonucleic acid, syntheses were required for recovery by the cells . The cells lost uracil- and leucine-labeled material as a result of the stress, further suggesting that ribonucleic acid and protein damage had occurred . Membrane damage was indicated by sensitivity to sodium dodecyl sulfate near the end of the lag period . Membrane damage was also suggested by the failure of cells to incorporate labeled material from the recovery medium . The lesions induced in this foodlike system are compared with those previously reported for a minimal media model system (Gray et al., Appl . Microbiol . 26:78-85, 1973; Gray et al., Appl . Environ . Microbiol . 33:1074-1078, 1977). J Dairy Sci, 1979 Mar, 62(3), 361 - 7 Thermal inactivation of a heat-resistant lipase produced by the psychotrophic bacterium Pseudomonas fluorescens; Andersson RE et al.; Lipase from Pseudomonas fluorescens was studied for thermostability at temperatures ranging from 100 C to 160 C . The heat treatments were in two media, and heating times necessary to inactivate 90% of the enzyme at constant temperature were extremely long even at high temperatures, e.g . 3.6 min at 140 C in nutrient broth and 2.0 min at 170 C in skim milk . The increments of temperature to reduce these heating times 90% were 37.0 C in nutrient broth and 38.9 C in skim milk . The lipase was inactivated only partly after 20 h at 20 C in 8 M urea, 6 M guanidine hydrochloride, and 1.0% sodium dodecyl sulfate . Four percent 2-mercaptoethanol showed no effect. C R Seances Soc Biol Fil, 1979, 173(4), 753 - 7 {Activation, by various aldoses, of dichlorophenol-indophenol reduction by endogenous constituents of a preparation of glucose dehydrogenase from Pseudomonas fluorescens}; Wurtz B; Dichlorophenol-indophenol is reduced neither by D-glucose nor by the endogenous components of a particulate purified glucose-dehydrogenase from Pseudomonas fluorescens, when these two classes of compounds acts individually . In contrast, the dye is quickly reduced by the endogenous components when the reaction occurs in the presence of glucose, without a direct participation of glucose in the reduction . In this effect D-glucose can be replaced by D-mannose, D-galactose or D-xylose, but not by D-fructose. Microbios, 1979, 24(95), 51 - 64 Cadmium and zinc sensitivity and tolerance in Bacillus subtilis subsp . niger and in a Pseudomonas sp; Pickett AW et al.; The action of Cd2+ and Zn2+ on Bacillus subtilis subsp . niger ATCC 9372, and on a Pseudomonas sp . (possibly Pseudomonas fluorescens), isolated from cadmium-polluted soil has been determined and compared with results obtained previously with Klebsiella aerogenes . In liquid medium the lag and the mean generation time of Bacillus subtilis subsp, niger increased with increasing Cd2+ or Zn2+ concentrations whereas only the total biomass of the Pseudomonas sp . was affected . Nevertheless, the responses of both species indicated a specific action at low concentrations and a more general toxic action at high concentrations . The survival on Cd2+ - or Zn2+ - agar depended on the state of the metal ions with regard to chelating ligands and on the nutritional stage of the organisms . In admixture, the metal ions acted synergistically, particularly on the Pseudomonas sp . Resistance to both metal ions developed . It was graded to the training concentration and reciprocal cross-resistance occurred with Bacillus subtilis . subsp . niger but not with the Pseudomonas sp. Biokhimiia, 1979 Jan, 44(1), 116 - 24 {Isolation and characterization of lipase from Pseudomonas fluorescens 533-5b}; Severina LO et al.; The enzyme was isolated from the culture fluid of Pseudomonas fluorescens 533-5b and purified by precipitation with (NH4)SO4 and acetone and by gel filtration through Sephadex G-200 . The enzyme was found homogeneous during polyacrylamide gel disc electrophoresis . The effects of metal ions, inhibitors, bile salts, temperature, pH and the substrate specificity of the enzyme were studied . It was shown that the enzyme from Ps . fluorescens 533-5b has a broad specificity . It can use as substrates many vegetable oils (olive, soybean, castor, sunflower, corn, mustard, linseed) . In addition, the enzyme is capable to hydrolyze synthetic triglycerides consisting of short-chained saturated fatty acids (butyric and caproic) and solid triglycerides containing saturated fatty acids with long carbon chains (myristic, lauric, stearic) . It is assumed that the enzyme is a glycoprotein; its molecular weight (320,000) and the amino acid composition were determined. Clin Chim Acta, 1978 Nov 1, 89(3), 447 - 53 An enzymic determination for serum phospholipid; Sugiura M et al.; A new colorimetric determination for serum phospholipid is described . Firstly, serum phospholipid is incubated with phospholipase C from Bacillus cereus, and then the released diglyceride and triglyceride are hydrolyzed completely to fatty acid and glycerol by lipoprotein lipase from Pseudomonas fluorescens . Secondly, the glycerol produced is enzymatically determined by glycerol dehydrogenase in the presence of NAD+, using phenazine methosulfate-nitro blue tetrazolium as color reagents . The absorbance at 570 nm is recorded . The amount of the glycerol from phospholipid is calculated by subtracting the amount of glycerol from triglyceride from the amount of total glycerol . The present method requires only 20 microliter of serum and a 40 min incubation and is highly reproducible . The results obtained show good correlation with those obtained by a chemical method (correlation coefficient, 0.925) or the phospholipase D-choline oxidase method (correlation coefficient, 0.936) . These results strongly suggest that the proposed method can be utilized as a routine clinical test. Prikl Biokhim Mikrobiol, 1978 Nov-Dec, 14(6), 858 - 65 {Isolation of the enzyme lipase from Pseudomonas fluorescens 533-5b and its characterization}; Bashkatova NA et al.; From the culture liquid of Pseudomonas fluorescens 533-5b, lipase (partially purified by sulphate ammonium precipitation and dialysis) was isolated . The following properties of the enzyme were examined: effect of pH and temperature, effect of bile salts, substrate specificity, and stability during storage . The optimal action of the preparation was at 55 degrees C and pH 7.5-8.0 . Sodium salts of cholic, taurocholic, deoxycholic, and glycocholic acids at concentrations over 0.5%, as a rule, activated lipolysis . The enzyme preparation was stable during storage: activity losses were no more than 15% during a 2 month storage at 4 degrees C . Lipase of Ps . fluorescens 533-5b was capable to utilize as substrates many vegetable oils (olive, corn, castor, mustard) as well as synthetic triglycerides containing carboxylic acids with short and long carbon chains. Biochim Biophys Acta, 1978 Sep 11, 526(1), 25 - 33 Purification of pantoate and dimethylmalate dehydrogenase from Pseudomonas fluorescens UK-1; Mantsala P; Pantoate dehydrogenase and dimethylmalate dehydrogenase were purified 69- and 112-fold, respectively, from Pseudomonas fluorescens UK-1 by ammonimu sulphate precipitation . Ultrogel AcA 34 gel filtration, hydroxyapatite column chromatography, heat treatment and Ultrogel AcA 44 gel filtration . The enzymes were evaluated for homogeneity (pantoate dehydrogenase was estimated to be about 95% pure) by disc and sodium dodecyl sulphate gel electrophoresis and by immunodiffusion . Pantoate and dimethylmalate dehydrogenases have molecular weights of 83 000 and 138 000, respectively, and are dissociable into four identical subunits with molecular weights of 24 000 and 34 000. Arch Microbiol, 1978 Aug 1, 118(2), 133 - 40 Effect of temperature on diauxic growth with glucose and organic acids in Pseudomonas fluorescens; Lynch WH et al.; Growth of Pseudomonas fluorescens in batch culture with glucose and organic acids resulted in typical diauxic responses at 30 degrees C but no detectable diauxic lag at 5 degrees C . At 30 degrees C, organic acids were preferentially utilized during the first growth phase . Glucose utilization was delayed until onset of the second growth phase . Systems involved in direct uptake and catabolism of glucose responded in a manner compatible with repression by malate and/or its metabolites and induction by glucose and/or its metabolites . The oxidative non-phosphorylated pathway, through gluconate and 2-ketogluconate (2-KG) as intermediates, was not induced during either growth phase . At 5 degrees C, growth with glucose and organic acids was biphasic but without diauxic lag . Organic acids were preferentially utilized during the first growth phase . Although carbon from glucose was not fully catabolized until onset of the second growth phase, glucose was oxidized to and accumulated extracellularly as gluconate and 2-KG during the first growth phase . No significant repression of glucose-catabolizing enzymes was observed during growth with organic acids in the presence of glucose . However, uptake activities for gluconate and 2-KG did not increase significantly until onset of the second growth phase . Thus, at low temperatures, psychotrophic P . fluorescens oxidized glucose to extracellular 2-KG, while growing on preferred carbon sources . The 2-KG was then catabolized after depletion of the organic acid. Mikrobiologiia, 1978 Jul-Aug, 47(4), 689 - 92 {Amino acid makeup characteristics of bacteria utilizing nonnatural compounds}; Naumova RP et al.; A peculiarity of the amino acid pool has been discovered for the first time in bacteria utilizing such synthetic compounds as gamma butyrolactam, epsilon-caprolactam and zeta-aminoenanthic acid . The main components of the amino acid pool are omega-aminoacids (including synthetic ones) as well as glutamic acid . The total amino acid content increases upon utilization of the compounds being tested three times (Pseudomonas fluorescens), five times (Pseudomonas dacunchae) and seven times (Pseudomonas perolens) as compared to the control variants . These data suggest that hydrolysis and transamination of synthetic omega-amino acids play an important role in the constructive and energy metabolism of the bacteria under study. J Bacteriol, 1978 Jul, 135(1), 18 - 28 F'-plasmid transfer from Escherichia coli to Pseudomonas fluorescens; Mergeay M et al.; Various F' plasmids of Escherichia coli K-12 could be transferred into mutants of the soil strain 6.2, classified herein as a Pseudomonas fluorescens biotype IV . This strain was previously found to receive Flac plasmid (N . Datta and R.W . Hedges, J . Gen Microbiol . 70:453-460, 1972) . ilv, leu, met, arg, and his auxotrophs were complemented by plasmids carrying isofunctional genes; trp mutants were not complemented or were very poorly complemented . The frequency of transfer was 10(-5) . Subsequent transfer into other P . fluorescens recipients was of the same order of magnitude . Some transconjugants were unable to act as donors, and these did not lose the received information if subcultured on nonselective media . Use of F' plasmids helped to discriminate metabolic blocks in P . fluorescens . In particular, metA, metB, and argH mutants were so distinguished . In addition, F131 plasmid carrying the his operon and a supD mutation could partially relieve the auxotrophy of thr, ilv, and metA13 mutants, suggesting functional expression of E . coli tRNA in P . fluorescens . In P . fluorescens metA Rifr mutants carrying the F110 plasmid, which carried the E . coli metA gene and the E . coli rifs allele, sensitivity to rifampin was found to be dominant at least temporarily over resistance . This suggests interaction of E . coli and P . fluorescens subunits of RNA polymerase . his mutations were also complemented by composite P plasmids containing the his-nif region of Klebsiella pneumoniae (plasmids FN68 and RP41) . nif expression could be detected by acetylene reduction in some his+ transconjugants . The frequency of transfer of these P plasmids was 5 X 10(-4). Mikrobiologiia, 1978 Jul-Aug, 47(4), 637 - 43 {Effect of the redox potential on the growth of aerobic microorganisms}; Andreeva EA et al.; The effect of redox potential was studied on the growth of the following aerobic microorganisms: Candida utilis, Bacillus megaterium, Pseudomonas fluorescens . The action of oxidizing agents (K3Fe(CN)6, KIO3, K2Cr207 and KMnO4) and reducing agents (ascorbic acid, sodium thioglycolate, K4Fe (CN)6 and Na2S2O3) on the growth rate was investigated . K3Fe(CN)6, ascorbic acid, sodium thioglycolate and K4Fe(CN)6 were found to be suitable for buffering the Eh of the medium . The potential could be shifted by 50--200 mV by adding various reducing and oxidizing agents . The rate of growth of C . utilis, Bac . megaterium and Ps . fluorescens did not depend on the potential value. Biochim Biophys Acta, 1978 Jun 2, 509(3), 519 - 36 Transport of alpha-aminoisobutyrate by cells and membrane vesicles of Pseudomonas fluorescens; Stephenson MC et al.; The transport of alpha-aminoisobutyrate into Pseudomonas fluorescens NCIB 8865 and membrane vesicles prepared from this organism has been studied . Uptake by cells was mediated by two active transport systems with different apparent Km values, while transport into membrane vesicles was mediated by a single component . The effect of inhibitors on the energy-coupling mechanism for alpha-aminoisobutyrate transport in these systems suggests that a membrane potential may play a significant role in supporting alpha-aminoisobutyrate transport . The magnitude of the membrane potential in the vesicle system, and the sensitivity of its generation to inhibitors, has been measured using 137Cs in the presence of valinomycin . Direct attempts to demonstrate a protonsymport mechanism for alpha-aminoisobutyrate transport were negative. J Bacteriol, 1978 Jun, 134(3), 861 - 74 Effects of low temperature on in vivo and in vitro protein synthesis in Escherichia coli and Pseudomonas fluorescens; Broeze RJ et al.; The effects of temperature on protein synthesis by Escherichia coli, a mesophile, and Pseudomonas fluorescens, a psychotroph, were investigated by using whole-cell and cell extract preparations . After shifts to 5 degrees C, protein was synthesized at a slowly decreasing rate for 1 h by both organisms, after which P . fluorescens synthesized protein at a new rate corresponding to its 5 degrees growth rate, in contrast to E . coli which did not synthesize protein at a measurable rate . In vitro protein-synthesizing systems using MS-2 RNA, endogenous mRNA, and purified polysomes were utilized to investigate initiation of translation at 5 degrees C . In these systems, P . fluorescens cell extracts synthesized protein at linear rates for up to 2 h at 5 degrees C, whereas E . coli cell extracts synthesized protein for only 25 min at 5 degrees C . The rates of polypeptide elongation, as tested by the incorporation of phenylalanine into polyphenylalanine by cell extract protein-synthesizing systems from both organisms, were identical over the range of 25 to 0 degrees C . The polysome profiles of E . coli whole cells shifted from 37 to 5 degrees C showed accumulation of 70S ribosomal particles and ribosomal subunits at the expense of polysomes . Similar experiements done with P . fluorescens resulted in polysome reformation at 5 degrees C . In vitro experiments demonstrated that the 70S ribosomal particles, which accumulated in E . coli at 5 degrees C, were capable of synthesizing protein in vitro in the absence of added mRNA . These in vivo and in vitro results suggest that incubation of E . coli at subminimal temperatures results in a block in initiation of translation causing polysomal runoff and the accumulation of 70S particles, some of which are 70S monosomes. Prikl Biokhim Mikrobiol, 1978 May-Jun, 14(3), 455 - 61 {Substrate specificity of lipase from Pseudomonas fluorescens}; Shabanova EA et al.; Substrate specificity of lipase isolated from the culture liquid filtrate of Pseudomonas fluorescens BKM-B-1151 was investigated with respect to vegetable oils and animal fats (olive, sunflower, cotton, mustard and soybean oils; beef and hog fats and their glycerides and fatty acid esters) . The preparation showed a high specificity to the quantitative composition of the reaction mixture (substrate: enzyme ratio), chemical structure of the substrate, and the emulgator type (gelatine, gum arabic and Triton X-100) . The lipase preparation hydrolyzed oils and water-insoluble fatty acid esters . The latter indicated an involvement of lipase. Mikrobiologiia, 1978 May-Jun, 47(3), 562 - 6 {Multiplication of Bdellovibrio bacteriovorus in the cytoplasm of the bacterial host}; Pechnikov NV et al.; The bacterial parasite Bdellovibrio bacteriovorus was studied in the process of its interaction with the host bacterium Pseudomonas fluorescens . As has been shown by time-lapse microcinematography, along with the normal growth of B . bacteriovorus in the periplasmatic space of the host bacterium, occasionally (4--5%) the parasite is located in the cytoplasm where the complete stage of its intracellular growth takes place with the release of progeny. Mikrobiologiia, 1978 Mar-Apr, 47(2), 234 - 40 {Exolipases of some Pseudomonas species}; Bashkatova NA et al.; The production of exolipase was studied in various Pseudomonas species, most of which did not possess the lipolytic activity in the conditions of the experiment, whereas in some strains this activity was rather high . The highest activity was displayed by the Pseudomonas fluorescens 533 strain and its mutant obtained upon UV irradiation . Lipase biosynthesis depended on the composition of a growth medium, the highest lipolytic activity being found on media with a high content of complex organic substances . No correlation was established between the growth and the lipolytic activity . A lipase preparation was isolated from the Ps . fluorescens 533-5b mutant, partly purified by precipitation with ammonium sulphate and dialysis, and its response to the action of inhibitors and heavy metal ions was studied . The enzyme activity was inhibited by these ions, and stimulated by Mg2+ ions . EDTA was found to be the strongest among all the inhibitors tested. Antibiotiki, 1978 Feb, 23(2), 122 - 5 {Biological properties of an asparaginase-glutaminase preparation from Pseudomonas fluorescens in cell cultures}; Kondrat'eva NA et al.; Specific L-asparaginase activity and non-specific cytotoxicity of asparaginase-glutaminase preparation from Pseudomonas fluorescens were studied . Two cell lines, i.e . the asparaginase-dependent (Berkitt lymphoma cells) and the asparaginase-independent (the ovary cancer cells) were used as the test-system . Incorporation of 3H-timidine into DNA was used as the criterion of the drug effect on the cells . Krasnitin was used as the reference preparation . The preparation of asparaginase-glutaminase was inferior to krasnitine by its specific antitumour asparaginase activity and superior to it by the general cytotoxicity in the cells of CaOv . With the help of the above test-system it is possible to study the specific asparaginase activity of the drugs containing L-asparaginase . For studying the specific glutaminase properties it is necessary to develop another cell test-system. C R Seances Soc Biol Fil, 1978, 172(4), 744 - 7 {Effect of dichlorophenolindophenol on glucose dehydrogenase activity of Pseudomonas fluorescens (type R)}; Wurtz B; The glucose dehydrogenase activity of Pseudomonas fluorescens cells grown in iron-depleted synthetic media is strongly decreased (about 80%) by dichlorophenolindophenol (DIP) 2 X 10(-3) M . In those cells, DIP seems not to play the part of an ultimate electron acceptor. Comp Biochem Physiol B, 1978, 61(1), 111 - 4 Comparative specificities of trehalases from various species; Labat-Robert J et al.; 1 . Using derivatives or non-symmetrical analogs of alpha,alpha-trehalose, we studied the catalytic specificities of trehalases from various species: Pseudomonas fluorescens, Melolontha vulgaris, porcine and human kidneys . 2 . alpha,Beta-trehalose, beta,beta-trehalose, 6,6'dideoxy alpha,alpha-trehalose, alpha-D-xylopyranosyl alpha-D-xylopyranoside were shown to be neither substrates nor inhibitors . 3 . 6'deoxy alpha,alpha-trehalose, alpha-D-glucopyranosyl alpha-D-xylopyranoside, alpha-D-allopyranosyl alpha-D-glucopyranoside and alpha-D-galactosyl alpha-D-glucopyranoside, which all possess an intact alpha-D-glucopyranosyl residue, were split by all these trehalases . 4 . alpha-D-glucopyranosyl alpha-D-mannopyranoside, alpha,alpha-trehalosamine are competitive inhibitors . 5 . These results show the importance of the primary alcohol group at C-6, of the equatorial configuration of the OH groups at C-2, C-3 and C-4 and of the modification of the structure at C-2 of the substrate for the catalytic activity. Z Allg Mikrobiol, 1978, 18(2), 135 - 41 Evaluation of yield and maintenance coefficients, expressed in carbon units, for Pseudomonas fluorescens and P . aeruginosa; Verstraete W et al.; The maximum cell yield (YMc - g biomass carbon per gram substrate carbon) and the rate of maintenance metabolism (mc - g substrate carbon/g biomass carbon per hour) have been determined for substrate limited continuous cultures of Pseudomonas fluorescens and P . aeruginosa . The metabolism of the organic substrates was monitored by measuring the COD-removal1) rates at different dilution rates . The advantages of expressing yield and maintenance coefficients in carbon units is discussed. C R Acad Sci Hebd Seances Acad Sci D, 1977 Dec 19, 285(16), 1537 - 40 {Effect of phosphorylated constituents on the growth of Pseudomonas fluorescens in media containing potassium oxalatoberyllate}; MacCordick J; Differences in the extent of beryllium inhibition on the growth of Pseudomonas fluorescens cultivated in various media are related to the concentration and nature of phosphorus-containing constituents of the substrate . Quantitative studies are carried out for interaction processes leading to reduction in beryllium toxicity . These include sequestration of beryllium in soluble form by beta-glycerophosphate and formation of sparingly soluble derivatives with inorganic orthophosphate ions. Arch Microbiol, 1977 Dec 15, 115(3), 271 - 5 Utilization of benzylpenicillin as carbon, nitrogen and energy source by a Pseudomonas fluorescens strain; Johnsen J; A bacterium which utilizes benzylpenicillin as carbon, nitrogen and energy source was isolated from a lake sediment . The organism was identified as a strain of Pseudomonas fluorescens with a GC content of 59.71 Mol% . After growth of the organism on a mineral salts medium containing benzylpenicillin, the derivatives benzylpenicilloic acid, benzylpenilloic acid and benzylpenicillenic acid were found in culture media . There was no indication that the phenylacetate side chain of benzylpenicillin is decomposed . In uninoculated culture media benzylpenicillin, benzylpenicilloic acid and benzylpenicillenic acid were demonstrable . The following compounds were found to be absent from inoculated or uninoculated culture fluids: D-penicillamine, L-valine, L-cysteine, benzylpenillic acid and 6-aminopenicillanic acid . The organism possesses penicillinase . Penicillin acylase was not demonstrable . The reaction product of penicillinase, benzylpenicilloic acid, supports only little growth . There is no growth on 6-aminopenicillanic acid with or without NH4Cl . Relatively little growth occurs on 6-aminopenicillanic acid in the presence of phenylacetic acid . The data indicate that the nucleus of the benzylpenicillin molecule is utilized as carbon, nitrogen and energy source . During growth a part of the substrate is destroyed into scarcely usable benzylpenicilloic acid; hereby the antibiotic is detoxified. Biochim Biophys Acta, 1977 Dec 8, 485(2), 424 - 33 Some properties of the extracellular protease produced by the psychotrophic bacterium Pseudomonas fluorescens strain AR-11; Alichanidis E et al.; The major extracellular protease from Pseudomonas fluorescens strain AR-11 has been partially purified by a factor of 300 by a combination of DEAE-cellulose ion-exchange chromatography and gel filtration . The enzyme had a molecular weight of 38 400 and exhibited optimum activity with isoelectrically precipitated casein substrate at pH 6.5 with Km - 0.13 mM . The protease was strongly inhibited by a number of heavy metal ions at the 10 mM level and also inhibited by thiol agents, while 10 mM EDTA led to slight activation . Optimum activity was retained, amounting to 33% of the maximum activity at 4 degrees C and 72% at 20 degrees C . Heat inactivation studies in which the isolated protease was heated at high temperature before subsequent incubation at 35 degrees C with substrate showed that for 50% inactivation 25 s heating at 130 degrees C or 17 s at 140 degrees C of 8.5 s at 150 degrees C was requried . The combination of high stability to heat treatments and retention of considerable activity at low incubation temperatures indicates that such a protease might have considerable significance in the processing and subsequent storage of food and other products. Biochim Biophys Acta, 1977 Nov 24, 489(2), 262 - 8 Physicochemical properties of a lipase from Pseudomonas fluorescens; Sugiura M et al.; The molecular weight of traicylglycerol lipase (EC 3.1.1.3) from Pseudomonas fluorescens is estimated to be approx . 33 000 by sodium dodecyl sulfate electrophoresis and Sephadex G-75 gel filtration . The lipase appears to be a single-chain protein and contains neither sugar nor lipid . The enzyme has a sedimentation coefficient (S20,w) of 3.06, an intrinsic viscosity of 3.0 g/ml and a partial specific volume of 0.730 g/ml, with an isoelectric point of pH 4.46 . Amino acid analysis showed that the enzyme contained few sulfur-containing amino acid residues with no disulfide links . The N-terminal residue of the enzyme was found to be alanine and optical rotation dispersion analysis showed that about 20% of the enzyme structure was in a helicla configuration. Mikrobiologiia, 1977 Nov-Dec, 46(6), 997 - 1002 {Metabolic products of hydrocarbon-oxidizing strains of Mycococcus lactis and Pseudomonas fluorescens and their influence on culture growth}; Spitsyna DN et al.; The effect of concentrations of organic acids in the cultural broth on growth was studied with the strains of Mycococcus lactis and Pseudomonas fluorescens oxidizing hydrocarbons . The ratio between the acid and neutral fractions in the cultural broth of Mycococcus lactis was also investigated as well as their action on the bacterial growth . Mycococcus lactis growing on paraffin was found to produce acid and neutral products at a ratio of 1:2 . The acid and neutral products of metabolism produced different action on the growth of the carbohydrate oxidizing bacteria . The acid products inhibited the growth at a concentration of organic acids in the medium above 500 mg/litre . The neutral products were partily assimilated by Mycococcus lactis. J Bacteriol, 1977 Nov, 132(2), 628 - 40 Chemotaxis by Bdellovibrio bacteriovorus toward prey; Straley SC et al.; A chemotaxis assay system that uses a modified Boyden chamber was characterized and used for measurements of chemotaxis by Bdellovibrio bacteriovorus strain UKi2 toward several bacterial species . Bacteria tested included both susceptible and nonsusceptible cells (Escherichia coli, Pseudomonas fluorescens, Bacillus megaterium, and B . bacteriovorus strains UKi2 and D) . None was attractive to bdellovibrios when present at densities below 10(7) cells per ml . Chemotaxis toward E . coli was studied most extensively; under conditions that minimized effects of osmotic shock to the cells, E . coli and exudates from E . coli at densities as high as 10(8) cells per ml failed to elicit a chemotactic response . Cell-free filtrates from mixed cultures of bdellovibrios and E . coli neither attracted nor repelled bdellovibrios . The data indicate that bdellovibrios do not use chemotaxis to locate prey cells. Arch Microbiol, 1977 Sep 28, 114(3), 281 - 6 Fructose metabolism in four Pseudomonas species; Van Dijken JP et al.; 1 . ATP-Dependent phosphorylation of fructose could not be detected in extracts of fructose-grown cells of Pseudomonas extorquens strain 16, Pseudomonas 3A2, Pseudomonas acidovorans and Pseudomonas fluorescens . Instead, phosphorylation of fructose to fructose-1-phosphate was found to occur when cell-free extracts were incubated with fructose and phosphoenolpyruvate . Such an activity could not be detected in cell-free extracts of succinate-grown cells . 2 . High levels of 1-phosphofructokinase were found in extracts of the above organisms when growth on fructose . 3 . Mutants of Pseudomonas extorquens strain 16 lacking 1-phosphofructokinase were unable to grow on fructose . Revertants to growth on fructose had regained the capacity to synthesize this enzyme, indicating its necessary involvement in fructose metabolism . 4 . A survey has been carried out of enzymes involved in carbohydrate metabolism in the species listed above. Biochim Biophys Acta, 1977 Sep 28, 488(3), 353 - 8 Purification, crystallization and properties of triacylglycerol lipase from Pseudomonas fluorescens; Sugiura M et al.; Triacylglycerol lipase of Pseudomonas fluorescens was purified from the crude enzyme by ammonium sulfate precipitation and chromatographies on Sephadex G-75 and DEAE-cellulose . The crystallization of the lipase was successfully carried out . The purified lipase was demonstrated to be homogenous on disc electrophoresis and its molecular weight was calculated to be 32 000 by gel filtration . The optimum pH for hydrolysis of sesame oil was 7.0 . The enzyme was stable up to 40 degrees C under the condition of pH 7.0 for 30 min and had more than 80% of the remaining activity between pH 5.0--11.0 at 37 degrees C for 60 min . The lipase was strongly inhibited by iodine and partially inhibited by FeCl3 and N-bromosuccinimide, and showed the most activity on tricaproyglycerol, among the triacylglycerols used. Vopr Med Khim, 1977 Sep-Oct, 23(5), 618 - 22 {Substrate specificity, inhibitors and kinetics of deamidase AG (asparaginase-glutaminase) from Pseudomonas fluorescens AG}; Kovalenko NA et al.; Deamidase AG (asparaginase-glutaminase) from Pseudomonas fluorescens AG was shown to hydrolyze 1-glutamine and 1-asparagine highly effectively . Besides, the enzyme exhibited the rather high rate of deamidation of D-asparagine and D-glutamine (70% and 100%, respectively), Nalpha-butyl asparagine (63%) and among peptides -- of glycyl-L-asparagine (40%) . L-glutamic acid gamma-methyl ester was hydrolyzed only slightly (5%) . Effect of several substrate analogues on the deamidase AG activity was studied as well . Albiciine (alpha-amino-beta-ureide propionic acid) proved to be the strongest inhibitor (100%) . Beta-Methyl aspartic acid, S-carbamoyl cysteine, alpha-ketoglutaric acid showed the slight inhibitory effect (20%) . Amount of active centres per enzyme molecule was estimated by means of 14C-albiciine . Deamidase AG had apparently only one active centre . In estimation of relationship between the rate of reaction and substrate (L-asparagine) concentration, the reaction was found to follow Michaelis-Menten kinetics, K(m) = 4.5 with 10-4 M. Arch Microbiol, 1977 Jul 26, 114(1), 87 - 9 Induction of polymyxin resistance in Pseudomonas fluorescens by phosphate limitation; Dorrer E et al.; Shift of Pseudomonas fluorescens NCMB 129 from a a phosphate rich into a phosphate limited medium results in a reduction of the membrane phospholipids phosphatidylethanolamine, phosphatidylglycerol and cardiolipin . Concomitantly a positively charged ornithine amide lipid is synthesized . The gradual increase of this lipid is paralleled by an increasing resistance to polymyxin B . The binding capacities of intact cells, and isolated inner and outer membranes for the antibiotic are reduced in the resistant organisms . It is discussed that the observed effect could be circumstantial evidence that the positively charged polymyxin B needs negatively charged receptors in biological membranes in order to exert its antibiotic activity. Vopr Med Khim, 1977 Jul-Aug, 23(4), 503 - 8 {Physico-chemical properties of deamidase AG from Pseudomonas fluorescens AG possessing antitumor activity}; Rakov SS et al.; Homogenous deamidase AG from Pseudomonas fluorescens AG was found to be a glycoprotein with molecular weight of about 13,000 daltons . The molecule consists apparently of four similar or identic subunits with molecular weight of about 30,000 daltons . The amino acid composition, N-terminal amino acids, the amount of chymotryptic peptides, containing 14C-carboxymethyl cysteine were studied . The enzyme exhibited distinct antitumoral effect on cells of Burkitt's lymphoma, sensitive to asparaginase, but did not exhibit marked cytotoxic action on cells of human ovarium cancer CaOV line, resistant to asparaginases. Mikrobiologiia, 1977 Jul-Aug, 46(4), 773 - 5 {Transport and excretion of a fluorescent pigment by cells of a Pseudomonas fluorescens culture}; Ukrainskii VV et al.; The transport and exretion of a yellow-green pigment were studied with the culture of Pseudomonas fluorescens K-1 . These processes were found to be accomplished by means of membranous formations having a lamellar structure. Mikrobiologiia, 1977 Jul-Aug, 46(4), 750 - 4 {Effect of formaldehyde on a Pseudomonas fluorescens strain}; Leonova VE et al.; The strain of Pseudomonas fluorescens was isolated from active ooze and was capable of growth on a medium containing 100 mg/litre of formaldehyde . As a result of stepwise selection, a variant of Ps . fluorescens 27 oxidizing 250 mg/litre of formaldehyde was obtained . It differed from the parent strain in cultural, morphological, and biochemical properties. Biochim Biophys Acta, 1977 Jun 10, 482(2), 453 - 60 Kynureninase-type enzymes and the evolution of the aerobic tryptophan-to-nicotinamide adenine dinucleotide pathway; Gaertner FH et al.; Kynureninase-type (L-kynurenine hydrolase, EC 3.7.1.3) activity has been found to be present in the livers of fish, amphibia, reptiles, and birds . In addition to past information concerning this enzyme activity in mammalian liver, it is now clear that all the major classes of vertebrates carry a highly specialized kynureninase-type enzyme, which we have termed a hydroxykynureninase . To compare the reactivities of these enzymes with L-kynurenine and L-3-hydroxykynurenine, ratios of tau values (Km/V) were used . Based on this comparison, the bacterium Pseudomonas fluorescens carries the most efficient kynureninase, whereas the amphibian Xenopus laevis has the most efficient hydroxykynureniase . In these two cases, the ratio of tau values differs by a factor of 38 000 . It is hypothesized that the tryptophan-to-nicotinamide adenine dinucleotide biosynthetic pathway evolved from a catabolic system of enzymes, and that the differences observed in the kynureninase-type enzymes between lower and higher organisms reflect the specialization of the function of these enzymes from a strictly catabolic role to an anabolic one during the course of evolution. J Med Chem, 1977 Jun, 20(6), 847 - 50 Synthesis and biological activity of some derivatives of thiochroman-4-one and tetrahydrothiapyran-4-one; Ramalingam K et al.; A small series of pyrazoles and isoxazoles derived from thiochroman-4-one has been synthesized and characterized . The compounds were examined for their in vitro inhibitory activity against Bacillus subtilis and Pseudomonas fluorescens . Among the tested compounds the pyrazole derivative from thiochroman-4-one was found to be the most effective inhibitor of growth of B . subtilis . Extensive H NMR analysis was recorded for all compounds. Appl Environ Microbiol, 1977 May, 33(5), 1074 - 8 Diluent sensitivity in thermally stressed cells of pseudomonas fluorescens; Gray RJ et al.; Thermally injured cells of Pseudomonas fluorescens were unable to produce colonies on Trypticase soy agar (TSA) after dilution with 0.1% peptone . Nutritional exigency could not be used as the criterion for this injury, since varying the composition of the plating medium had little effect on the number of colonies that developed . The injured cells had no requirement for compounds known to leak out during the heat treatment in order to recover . The cells did not exhibit injury if dilution preceded heat treatment on the plating medium, demonstrating that the heat treatment sensitized the cells to the trauma of dilution . Substitution of 0.1% peptone with growth medium as the diluent largely offset the previously observed drop in TSA count . Little difference in survival was observed when monosodium glutamate or the balance of the defined medium was used as the diluent . The diluent effect was ionic rather than osmotic . The presence of cations was important in maintaining the integrity of the injured cell, and divalent cations enhanced this protective effect . The role of these cations at the level of the cell envelope is discussed. J Med Chem, 1977 May, 20(5), 664 - 9 Synthesis and antimicrobial activity of azasteroid-type compounds and related systems . Effect of hydrophilic and lipophilic groups on activity; Brown RA et al.; Pyrazole-, pyrazolone- and isoxazole-containing systems were prepared from 3,4-dihydro-6-(hexyloxy)-1(2H)-naphthalenone, 3,4-dihydro-6-(hexadecyloxy)-1(2H)-naphthalenone,3,4-dihydro-6(2-dimethylaminoethyloxy)-1-(2H)-naphthalenone, 3,4-dihydro-7-hexyloxy-1(2H)-phenanthrone, and 3,4-dihydro-7-(2-dimethylaminoethyloxy)-1(2H)-phenanthrone . A number of compounds derived from 7, 8-dihydro-5(6H)-quinolinone were also synthesized and characterized . Both hydrophilic and lipophilic groups were incorporated into certain systems as well as cidal groups . The compounds were screened for their in vitro inhibitory activity against Bacillus subtilis and Pseudomonas fluorescens . Structure-acitivity relationships among the molecular systems are discussed. Eur J Biochem, 1977 Apr 15, 74(3), 441 - 5 4-amino-hex-5-enoic acid, a selective catalytic inhibitor of 4-aminobutyric-acid aminotransferase in mammalian brain; Lippert B et al.; Incubation of rat brain 4-aminobutyrate aminotransferase with 4-amino-hex-5-enoic acid, a substrate analog of 4-aminobutyric acid, results in a time-dependent irreversible loss of enzymatic activity . In the presence of 0.1 mM inhibitor the half-life of the inactivation process is approximately 6 min . Low concentrations of L-glutamic acid or 4-aminobutyric acid protect against this inactivation, while 2-oxoglutarate prevents this protection, suggesting that only the pyridoxal form of the enzyme is susceptible to inhibition by 4-amino-hex-5-enoic acid . The irreversible inhibition of mammalian 4-aminobutyrate aminotransferase by 4-amino-hex-5-enoic acid is selective . There is no inhibition of this enzyme from Pseudomonas fluorescens with the inhibitor at mM concentrations . Even at 10 mM there is no irreversible inhibition of mammalian glutamate decarboxylase or of aspartate aminotransferase, while alanine aminotransferase is inhibited over 500 times more slowly than rat brain 4-aminobutyrate transaminase. J Bacteriol, 1977 Apr, 130(1), 154 - 9 Role of the 30S ribosomal subunit, initiation factors, and specific ion concentration in barotolerant protein synthesis in Pseudomonas bathycetes; Landau JV et al.; Washed (1 M NH4Cl) ribosomes from Pseudomonas bathycetes, Pseudomonas fluorescens, and Escherichia coli were tested for their ability to synthesize protein or polypeptide at high pressure when used as such, when recombined with homologous initiation factors, and when recombined with heterologous initiation factors . The responses of natural messenger ribonucleic acid (MS-2)-directed systems to pressure were independent of the source of initiation factors and paralleled those of the washed ribosomes in polyuridylate-directed systems . In all cases, the responses to pressure were parallel to those obtained when unwashed ribosomes were utilized; therefore, we concluded that the initiation factors were interchangeable among these organisms, and that these factors did not play a critical role in determining the pressure responses of the protein-synthesizing systems . P . bathycetes ribosomal subunits were isolated under a variety of ionic conditions . These were tested for their ability to synthesize protein and polyphenylalanine at a variety of pressures when used in reconstituted P . bathycetes homologous systems and in hybrid systems with ribosomal subunits from E . coli and P . fluorescens . O . bathycetes 30S subunits, isolated in a buffer solution containing 0 mM NaCl and O mM KC} were functional at any pressure; those isolated in the presence of 150 mM NaCl and 0 mM KCl were functional at 1 atmosphere but barosensitive, and those isolated in the presence of O mM NaCl and 150 mM KCl retained the ion-mediated barotolerance characteristic of crude P . bathycetes ribosome preparations . The 50S subunit remained functional regardless of the method of isolation, and it had no effect on pressure sensitivity. Biochemistry, 1977 Mar 8, 16(5), 895 - 901 Tyrosine emission in the tryptophanless azurin from Pseudomonas fluorescens; Ugurbil K et al.; A strain of Pseudomonas fluorescens contains an azurin with no tryptophan and two tyrosines . This protein is interesting because it allows one to study both the structure of azurin and the emission of tyrosines in proteins . Comprehensive measurements were carried out including spectrophotometric and fluorimetric titration, fluorescence quantum yield, fluorescence polarization, and I- quenching . In the copper-containing protein, almost independent of the copper ion oxidation, the fluorescence quantum yield is approximately 60% of that of the apoprotein . The latter has the remarkable property that its quantum yield is even greater than free tyrosine . The two tyrosines in the metalloprotein have different pKa's, 10.75 and 12.78, but there is only one average pKa, 10.9 in the apoprotein . The polarization of the fluorescence at 310 nm (290-nm excitation) is 0.32 for the metalloproteins and 0.34 for the apoprotein . I- hardly quenches the fluorescence . The conclusion is that the two tyrosines are inaccesible to the solvent, located in nonpolar environments, larger than or equal to 20 A apart, and not adjacent to the disulfide bridge. J Bacteriol, 1977 Mar, 129(3), 1365 - 74 p-Cymene pathway in Pseudomonas putida: ring cleavage of 2,3-dihydroxy-p-cumate and subsequent reactions; DeFrank JJ et al.; It was confirmed that 2,3-dihydroxy-p-cumate is a substrate for ring cleavage in Pseudomonas putida PL-W after growth with p-cymene or p-cumate . This compound was oxidized to pyruvate, acetaldehyde, isobutyrate, and carbon dioxide by extracts of cells, and these products appear in equimolar amounts . The transient appearance of compounds and 2,3-dihydroxy-p-cumate to a yellow intermediate (lambda max, 345 nm) without decarboxylation . Extracts of the benzene nucleus; this is followed by decarboxylation to give the 393-nm species, which gives rise to isobutyrate, acetaldehyde, and pyruvate by the hydrolytic route of meta cleavage of catechols, via 4-hydroxy-2-oxovalerate . This was confirmed with a mutant of P . putida PL-RF-1 that was unable to grow with p-cymene (or p-cumate) but was able to oxidize both compounds AND 2,3-DIHYDROXY-P-CUMATE TO A YELLOW INTERMEDIATE (LAMBDA MAX, 345 NM) WITHOUT DECARBOXYLATION . Extrats of P . putida PL-W (wild type) or a revertant of the mutant PL-RF-1 catalyzed the decarboxlation of the 345-nm intermediate with transient formation of the compound that absorbed at 393 nm . The substrate specificities of the 3,4-dioxygenative ring cleavage enzyme, and the decarboxylase were determined in crude extracts of P . putida PL-W and Pseudomonas fluorescens 007 . It was conclude that 3,4-dioxygenative cleavage and decarboxylation are sequential enzyme-catalyzed reactions common to both P . putida and P . fluorescens for the oxidation of 2,3-dihydroxybenzoates . Unlike P . putid