J Biochem (Tokyo), 1975 Aug, 78(2), 247 - 51 Studies on the subsite structure of amylases . II . Difference-spectrophotometric studies on the interaction of maltotriose with liquefying alpha-amylase from Bacillus subtilis; Ohnishi M et al.; The difference spectra of liquefying alpha-amylase (EC 3.2.1.1) from B . subtilis upon the addition of a slowly reacting substrate, maltotriose, were measured to investigate specific binding of the substrate to the enzyme . The spectra produced by maltotriose were attributed to one tryptophan and one tyrosine residues on the basis of analysis of their shape and magnitude . From the dependence of the difference absorption upon the concentration of maltotriose, the dissociation constant of the maltotriose-enzyme complex was determined to be 170(+/- 20) mM, which is in good agreement with the Michaelis constant, Km obtained from the steady-state kinetics . The difference spectrum characteristic of a tryptophan residue was significantly decreased by the chemical modification of a trytophan residue with N-bromosuccinimide. J Antibiot (Tokyo), 1975 Aug, 28(8), 567 - 72 Antraformin, a new inhibitor of Bacillus subtilis transformation; Tanaka T; A convenient method was worked out which discriminates between inhibitory activity of compounds against Bacillus subtilis transformation and their antibacterial or growth inhibition activity . By this assay system, several drugs and antibiotics were tested and some of them were found to be more inhibitory to transformation than to bacterial growth . This method was further applied to look for specific inhibitors among culture broths of Streptomyces, and during this screening program, Streptomyces sp . 7725-CC1 was found to produce a specific inhibitor of B . subtilis transformation . The active substance was purified and named antraformin after its specific action . The molecular weight was suggested to be 279 (C14H21N3O2) for the compound by high resolution mass spectrometry. J Virol, 1975 Aug, 16(2), 315 - 21 N-Glycosidase activity in extracts of Bacillus subtilis and its inhibition after infection with bacteriophage PBS2; Friedberg EC et al.; We have detected in crude extracts of Bacillus subtilis an N-glycosidase activity which catalyzes the release of free uracil from DNA of the subtilis phage PBS2 labeled with {3H}uridine . This DNA contains deoxyuridine instead of thymidine . The enzyme is active in the presence of 1.0 mM EDTA and under these conditions Escherichia coli or T7 DNA labeled with {3H}thymidine is not degraded to labeled acid-soluble products . The activity resembles an N-glycosidase from E . coli which releases free uracil from DNA containing deaminated cytosine residues . Both enzymes in crude extracts are active in the presence of EDTA, do not require dialyzable co-factors, and have the same pH optimum . They differ in that the enzyme from E . coli is more sensitive to heat, sulfhydryl reagents, and salt . The enzyme from B . subtilis is inactive on DNA containing 5-bromouracil or hydroxymethyluracil . Extracts of PBS2-infected B . subtilis lose the N-glycosidase activity within 4 min after infection and contain a factor that inhibits the N-glycosidase activity within 4 min after infection and contain a factor that inhibits the N-glycosidase activity in extracts of uninfected cells in vitro. J Pharm Sci, 1975 Aug, 64(8), 1305 - 8 Chemical and pharmacological investigations of constituents of Eleutherine bulbosa (Miller) Urb . (Iridaceae); Bianchi C et al.; Eleutherin and eleutherol extracted from bulbs of Eleutherine bulbosa (Miller) Urb . (Iridaceae), collected in the Amazonian jungle and grown in Italy, were tested for biological properties . The extraction procedure and the results of antibacterial, cytotoxicity, and pharmacological assays are reported . Eleutherin has a weak and transient effect of decreasing the prothrombin time (in vivo in rats) and a weak antibacterial activity on Bacillus subtilis (in vitro). J Bacteriol, 1975 Aug, 123(2), 604 - 15 Pyrimidine biosynthetic pathway of Baccillus subtilis; Potvin BW et al.; Biochemical and genetic data were obtained from a series of 51 Pyr- strains of Bacillus subtilis . The observed enzymatic deficiencies allowed the mutants to be placed into 12 clases, some of which represent defects in more than one of the six known pyrimidine biosynthetic enzymes . Mapping analysis by transformation has shown that all the Pyr- mutations are located in a single small area of the B . subtilis genome . A correlation of the biochemical defects and the genetic data has been made . Those mutations conferring similar enzymatic deficiencies were found to be contiguous on the B . subtilis map . Regulatory aspects of the pyrimidine pathway have also been investigated and are compared to previously reported results from other organisms . Evidence is presented which bears upon the possible physical association of the first three enzymes and the association of at least some of the enzymes of this pathway with particulate elements of the cell . A model for the organization of the enzymes is presented with dihydroorotate dehydrogenase as the central enzyme in a proposed aggregate. J Biochem (Tokyo), 1975 Aug, 78(2), 381 - 90 Pyridine-2, 6-dicarboxylic acid (dipicolinic acid) formation in Bacillus subtilis . II Non-enzymatic and enzymatic formations of dipicolinic acid from alpha, epsilon-diketopimelic acid and ammonia; Kimura K et al.; Non-enzymatic formation of dipicolinic acid (DPA) from diketopimelic acid and ammonia was clearly demonstrated using a new method for DPA analysis . The reaction rates of DPA formation were almost the same under aerobic and anaerobic conditions . Nearly equimolecular quantities of DPA and tetrahydrodipicolinic acid were detected in spontaneous reaction mixture . The spontaneous reaction seemed to be due to dismutation of dihydrodipicolinic acid, resulting in DPA and tetrahydrodipicolinic acid . The apparent optimum pH of the spontaneous reaction was 8.2 and the maximal rate of DPA formation was observed with a 1 : 4 molar ratio of diketopimelic acid to ammonia . The rate of the spontaneous reaction was stimulated by ferrous sulfate, FMN, and riboflavin . Dihydrodipicolinate reductase catalyzes the reduction of dihydrodipicolinate, prepared from pyruvate and aspartic beta-semialdehyde, with NADPH as reductant . The reductase was isolated from Bacillus subtilis, and found to stimulate DPA formation from diketopimelic acid and ammonia . The enzymatic DPA formation was absolutely dependent on oxygen, and optimum pH was 6.4 . The catalytic action of the enzyme was similar to that of the oxidase . Possible mechanisms of DPA formation from diketopimelic acid and ammonia are proposed. Biochim Biophys Acta, 1975 Jul 27, 397(1), 5 - 8 Stereochemistry of the hydrogen transfer to NAD catalyzed by (S)alanine dehydrogenase from Bacillus subtilis; Alizade MA et al.; The stereochemistry of the hydrogen transfer to NAD catalyzed by (S)alanine dehydrogenase { (S)alanine: NAD oxidoreductase (EC 1.4.1.1) } from B . subtilis was investigated . The label at C-2 of (S) {2,3--3H} alanine was enzymatically transferred to NAD, and the {4--3H}NADH produced isolated and the stereochemistry at C-4 investigated . It was found that the label was exclusively located at the (R) position which indicates that (S)alanine dehydrogenase is an A-type enzyme . This result was confirmed in an alternate way by reducing enzymatically {4--3H}NAD with non labeled (S)alanine and (S)alanine dehydrogenase and investigating the stereochemistry of the }4--3H}NADH produced . As expected, the label was now exclusively located at the (S) position . This proves that (S)alanine dehydrogenase isolated from B . subtilis should be classified as an A-enzyme with regard to the stereochemistry of the hydrogen transfer to NAD. Mol Gen Genet, 1975 Jul 10, 138(4), 299 - 307 Ribosomal proteins of Bacillus subtilis vegetative and sporulating cells; Guha S; The ribosomal subunit proteins(30S and 50 S) from vegetative and sporulating cells of Bacillus subtilis 168M were analyzed by two dimensional acrylamide gel electrophoresis . Twenty two proteins were identified in the 30S subunits and 28 proteins are detectable in the 50S subunits . The number of proteins and their electrophoretic mobility seem to remain unaltered during the sporualtion process . The ribosomal proteins of a thermosensitive sporulation mutant (ts-4), isolated from stationary phase cultures, under permissive (for sporulation) and non-permissive conditions, did not show any qualitative difference in either of the subunits . The 21S precursor particles derived from log phase cell ribosomes show two different proteins, in addition to those present in the 30 S subunit . It is suggested that these two proteins either disappear or are modified during the maturation process. Mol Gen Genet, 1975 Jul 10, 138(4), 269 - 79 In vitro modification and restriction of phage phi-105c DNA with Bacillus subtilis N cell-free extract; Shibata T et al.; The enzymes involved in host-controlled modification and restriction by Bacillus subtilis strain N were detected in cell free extracts . In the presenct of Mg2+ the N-specific endonucleases cleaved unmodified DNA but did not attack phi-105C . N DNA carrying N-specific modification . The restriction endonuclease required neither SAM nor ATP for its activity . The N-specific modification enzyme was active only in the presence of SAM, indicating that modification in this syteem is a methylation of DNA. Biochim Biophys Acta, 1975 Jul 7, 395(3), 294 - 305 Studies on DNA repair in Bacillus subtilis . II . Partial purification and mode of action of an enzyme enhancing the priming activity of gamma-irradiated DNA; Noguti T et al.; A cellular factor which makes T7 DNA irradiated with gamma-rays a better primer for Micrococcus DNA polymerase was partially purified by DEAE and phosphocellulose column chromatography and named "primer activating enzyme" . Sucrose density gradient sedimentation analysis was carried out to examine actions of one major active fraction that appeared by phosphocellulose chromatography . It was shown that this factor introduced new nicks in T7 DNA in addition to those introduced directly by gamma-ray irradiation . This enzyme fraction also had an endonucleolytic activity towards DNA containing apurinic sites induced by heat treatment and had capacity to enhance the priming activity of heat- or methyl methansulfonate-treated DNA but affected very little that of ultraviolet-irradiated DNA . This enzyme had no effect on T7 DNA when it was not treated with the DNA-damaging agents . From these results we concluded that this enzyme may be analogous to the endonuclease II or apurinic site-specific enconuclease of Escherichia coli. Biochim Biophys Acta, 1975 Jul 7, 395(3), 284 - 93 Studies on DNA repair in Bacillus subtilis . I . A cellular factor acting on gamma-irradiated DNA and promoting its priming activity for DNA polymerase I; Noguti T et al.; Initiation of new DNA synthesis was observed in B . subtilis cells upon gamma-ray irradiation followed by toluene treatment and incubation in the presence of the four deoxynucleotide triphosphates and Mg2+ . This DNA synthesis took place in the absence of ATP and was refractory to 6-(p-hydroxyphenylazo)-uracil which is a specific inhibitor for the type III polymerase of Bacillus subtilis . This repair-type DNA synthesis was greatly reduced in mutant cells deficient in DNA polymerase I . Restoration of transforming activity of cellular DNA was found to occur in parellel with the above repair type DNA synthesis . A protein factor which enhances the priming activity of gamma-irradiated DNA for DNA polymerase I was detected in DNA-free extracts prepared from B . subtilis cells by means of lysis with a buffer containing lysozyme, Brij-58 and EDTA. Mikrobiologiia, 1975 Jul-Aug, 44(4), 727 - 31 {Characteristics of spore-forming bacteria of the genus Bacillus that break down caprolactam}; Rotmistrov MN et al.; Five strains of sporeforming bacteria were isolated from sewage of capron industry and their morphological, cultural, as well as physiological and biochemical properties were investigated . Four strains were identified as Bacillus subtilis and one as Bacillus pumilus . The cultures were able to grow in mineral medium with caprolactam as a source of carbon and nitrogen . The influence of growth conditions on the rate of caprolactam decomposition in synthetic medium was studied. J Pharm Sci, 1975 Jul, 64(7), 1240 - 2 Synthesis, spectral properties, and antibacterial activity of synthetic precursors of macrocyclic oxa- and thia-substituted benzolactones and benzoketones; Jones PR et al.; Terminally difunctional compounds were synthesized by alkylation of salicylic acid, thiosalicylic acid (o-mercaptobenzoic acid), or their derivatives . Whereas methyl salicylate and thiosalicylic acid were smoothly etherified, salicylic acid was alkylated at the carboxyl . Characteristic IR and NMR spectral patterns in the synthesized compounds can be attributed to intramolecular hydrogen bonding or substituent effects and are consistent with observations already reported for similar compounds . Three synthesized compounds exhibited low but reproducible inhibitory effects on Bacillus subtilis growth. J Pharm Sci, 1975 Jul, 64(7), 1224 - 9 Microbiological diffusion assay II: design and applications; Kavanagh F; Application of new equipment and new techniques was made to antibiotic diffusion assays . Accumulation of data and computation of potencies were made by an on-line computer . The system was tested by assaying cephalexin with the aid of Bacillus subtilis in an FDA single-dose design modified by reducing the number of standards to two . The influence of the thickness of the base layer and the form of the dose-response line were tested . Zone diameter was measured with a resolution of 0.01 mm . The potency of samples was measured with an error usually less than 3% . An error of 0.1 mm in measuring zone size would cause an error of 3% of potency . The usual calibration line was inadequate for extrapolation beyond a twofold range . A dose-response line derived from the Cooper equation was better for standard curves spanning more than a twofold range of concentrations . Precision was better on the plates with the 20-ml base layer . The two-dose method of assaying gave larger errors than the single-dose method . Large variances in measuring zone diameters are a reason for the low precision of diffusion assays and set an inherent limit on precision. J Antibiot (Tokyo), 1975 Jul, 28(7), 543 - 9 Lipiarmycin, a new antibiotic from Actinoplanes III . Mechanism of action; Sergio S et al.; In vivo, at low concentrations (less than or equal to 1 mug/ml), the antibiotic lipiarmycin specifically inhibits RNA synthesis in Bacillus subtilis . At a much higher concentration (100 mug/ml), syntheses of other macromolecules such as DNA and protein also appear to be suppressed . In vitro, the antibiotic caused 50% inhibition of DNA-dependent RNA-polymerase from B . subtilis at a concentration of 0.6 mug/ml and of that E . coli at 5 approximately 8 mug/ml . The activity of Escherichia coli DNA-polymerase I is inhibited 50% at 55 approximately 65 mug/ml . Lipiarmycin prevent ribonucleoside triphosphate polymerization only if added prior to the association between RNA-polymerase and DNA, and does not affect the elongation rate of RNA chains at concentrations up to 100 mug/ml . At that concentration, however, the antibiotic immediately blocks the polymerization of deoxyribonucleotide triphosphates catalyzed by DNA-polymerase I. Can J Microbiol, 1975 Jul, 21(7), 1129 - 32 Ultraviolet sensitivity and photoproducts in spore-like bodies of an excision-repair-deficient and dipicolinic-acid-less strain of Bacillus subtilis; Munakata N et al.; Bacillus subtilis strain UVS-42DPA is defective in both excision-repair capability and dipicolonic acid(DPA)accumulation . In sporulation medium, it forms spore-like bodies, which are sensitive to ultraviolet light (UV) as the vegetative cells and produce mostly cyclobutane dimers instead of "spore photoproduct" upon UV irradiation . The results suggest that the drastic change in the photochemical reactivity of DNA during sporulation might be induced and(or) maintained by the accumulation of DPA. J Bacteriol, 1975 Jul, 123(1), 83 - 95 Isolation, characterization, and mapping of Bacillus subtilis 168 germination mutants; Trowsdale J et al.; After mutagenesis with nitrosoguanidine, germination mutants of Bacillus subtilis 168 were selected by killing, with heat, spores that germinated at 42 C and collecting survivors at 30 C . The germination properties of nine mutants variously affected in amino acid biosynthesis and sugar utilization were studied in detail . They were divided into two groups: (i) Ger-ALA mutants, failed to germinate in 10 mM L-alanine but germinated in complex media (some of these mutants were temperature sensitive); (ii) Ger-PAB mutants, germinated poorly, even in complex media, suggesting that they were blocked in important germination functions . All the mutants failed to germinate in L-alpha-amino-n-butyrate or L-valine (including temperature-sensitive mutants only at the restrictive temperature) showing that there is a step necessary for germination affected by all three acids . The mutants had normal growth rates, indicating that the defective gene products were specific for germination functions . These defects were not identified . Eight of the mutants were mapped by transduction with phage PBS-1 . The recombinants were scored either by observations, by microscopy of phase darkening of the spores, or by a plate test involving the reduction of tetrazolium by heated colonies of spores . Five of the mutations, of at least three phenotypes, were between thr-5 and cysB3 away from all the sporulation markers that have been previously mapped . A linked ald (alanine dehydrogenase) locus was on the other side of thr-5 . The other Ger markers were located in at least two additional positions . Auxotrophic strains that were used for mapping germinated normally, but germination of the Ger mutants differed slightly in different genetic backgrounds. J Bacteriol, 1975 Jul, 123(1), 7 - 19 Control of cell length in Bacillus subtilis; Sargent MG; During inhibition of deoxyribonucleic acid synthesis in Bacillus subtilis 168 Thy-minus Tryp-minus, the rate of length extension is constant . A nutritional shift-up during thymine starvation causes an acceleration in the linear rate of length extension . During a nutritional shift-up in the presence of thymine, the rate of length extension gradually increases, reaching a new steady state at about 50 min before the new steady-state rate of cell division is reached . The steady-state rates of nuclear division and length extension are reached at approximately the same time . The ratio of average cell length to numbers of nuclei per cell in exponential cultures is constant over a fourfold range of growth rates . These observations are consistent with: (i) surface growth zones which operate at a constant rate of length extension under any one growth condition, but which operate at an absolute rate proportional to the growth rate of the culture, (ii) a doubling in number of growth zones at nuclear segregation, and (iii) a requirement for deoxyribonucleic acid replication for the doubling in a number of sites. J Bacteriol, 1975 Jul, 123(1), 366 - 71 Motility of Bacillus subtilis during growth and sporulation; Nishihara T et al.; The change of motility and the presence of flagella were followed throughout growth and sporulation in a standard sporulating strain and in 19 cacogenic sporulation mutants of Bacillus subtilis . For the standard strain, the fraction of motile cells decreased during the developmental period to less than 10% at T4 . Motility was lost well before the cells lose their flagella . Conditions reducing the decrease of motility also reduced sporulation: motile cells never contained spores . The decrease of motility was not coupled with a decrease in the cellular concentration of adenosine 5'-triphosphate or a decline in oxygen consumption, but an uncoupling agent immediately destroyed motility at any time . Apparently, motility decreased during development because it became increasingly uncoupled from the energy generating systems of the cell . The motility of sporulation mutants decreased after the end of growth at the same time as or earlier than the motility of the standard strain; the early decrease of motility in an aconitase mutant, but not that in an alpha-ketoglurate dehydrogenase mutant, could be avoided by addition of L-glutamate . Sporulation or related events such as extracellular antibiotic or protease production were not needed for the motility decline. J Bacteriol, 1975 Jul, 123(1), 346 - 53 Characterization of a rifampin-resistant, conditional asporogenous mutant of Bacillus subtilis; Murray CD et al.; A rifampin-resistant, conditionally asporgoenous mutant of Bacillus subtilis was isolated that sporulates poorly in Sterlini-Mandelstam sporulation medium, but that sporulates normally in modified Difco sporulation medium . Rifampin-resistant (Rif-r) and conditional asporogenous (Spo-c) phenotypes co-transformed at 100% frequency . Preliminary genetic studies indicated the Rif-r trait to lie between cysA14 and ery, a locus (rnp) common to Rif-r mutants . Ribonucleic acid polymerase from strains bearing this mutation was found to be rifampin resistant in vitro . The loss of ability to sporulate in Sterlini-Mandelstam medium was found to be corrected, to a large extent, by addition to the medium of arginine, methionine, valine, and isoleucine . Several other amino acids had small effects, whereas others had no effect at all . The restorative effect is approximately additive . Growth studies indicated that Rif-r strains grew more rapidly than the corresponding parent in minimal medium at temperatures higher than 37 C . Addition of certain amino acids to the medium resulted in identical growth rates at these temperatures . Extracellular protease and esterase activities of the Rif-r Spo-c mutant were normal . A slight difference was found in the heat sensitivity of partially purified ribonucleic acid polymerase preparations of this mutant compared to the wild type. J Virol, 1975 Jul, 16(1), 184 - 91 Isolation and characterization of prophage mutants of the defective Bacillus subtilis bacteriophage PBSX; Thurm P et al.; Bacillus subtilis mutants with lesions in PBSX prophage genes have been isolated . One of these appears to be a regulatory mutant and is defective for mitomycin C-induced derepression of PBSX; the others are defective for phage capsid formation . All of the PBSX structural proteins are synthesized during induction of the capsid defective mutants; however, several of these proteins exhibit abnormal serological reactivity with anti-PBSX antiserum . The two head proteins X4 and X7 are not immunoprecipitable in a mutant which fails to assemble phage head structures . In the tail mutant, proteins X5 and X6 are not immunoprecipitable, tails are not assembled, and a possible tail protein precursor remains uncleaved . The noninducible mutant does not synthesize any PBSX structural proteins after exposure to mitomycin C . The mutation is specific for PBSX since o105 and SPO2 lysogens of the mutant are inducible . All of the known PBSX-specific mutations were shown to be clustered between argC and metC on the host chromosome . In addition, the metC marker was shown to be present in multiple copies in cells induced for PBSX replication . This suggests that the derepressed prophage replicates while still integrated and that replication extends into the adjacent regions of the host chromosome. J Virol, 1975 Jul, 16(1), 179 - 83 Bacteriophage-specific protein synthesis during induction of the defective Bacillus subtilis bacteriophage PBSX; Thurm P et al.; Particles of PBSX, a defective, noninfectious phage which is inducible from strains of Bacillus subtilis 168, contain at least seven structural proteins resolvable by sodium dodecyl sulfate polyacrylamide gel electrophoresis . Five of these proteins are associated with the phage tail and two with the phage head . An eighth protein, which also may be coded for by the PBSX prophage, has been identified in cells derepressed for PBSX replication. Appl Microbiol, 1975 Jul, 30(1), 1 - 3 Effect of pH on sporulation of Bacillus stearothermophilus; Yazdany S et al.; An improved broth medium was developed for high growth yields of Bacillus subtilis var . niger NCIB 8649, Bacillus cereus NCIB 9373, and Bacillus stearothermophilus NCIB 8919 and ATCC 7953 . Sporulation was abundant (1.1 times 10-8 B . subtilis var . niger and 9.2 times 10-7 B . cereus per ml) at an initial pH of 7.0 . Sporulation of both strains of B . stearothermophilus took place (1.9 times 10-7 and 2.4 times 10-7/ml, respectively) in this medium when initial pH values of 7.7 to 8.7 were used. Z Immunitatsforsch Exp Klin Immunol, 1975 Jul, 149(2-4), 126 - 35 Estimates of the porosity of Bacillus licheniformis and Bacillus subtilis cell walls; Hughes RC et al.; The maximum porosity of Bacillus subtilis and Bacillus licheniformis cell walls was estimated by two independent and relatively simple methods . Peptidoglycan was isolated from Bacillus subtilis cell wall preparations and used as an insoluble support for exclusion chromatography of dextrans of known average molecular size . In an alternative approach the leakage of radioactively labelled proteins from Bacillus licheniformis cells incubated in butanol-saline mixtures was measured and their size estimated by exclusion chromatography on Sephadex G-100 . Under these conditions the permeability barrier of the cytoplasmic membrane was destroyed with preservation of the structural integrity of the outer cell wall . The apparent exclusion threshold of the cell wall of either organism as determined by these means corresponded to molecules with a diffusional radius of not more than 2.5 nm. J Bacteriol, 1975 Jul, 123(1), 123 - 7 Active transport of manganese in isolated membrane vesicles of Bacillus subtilis; Bhattacharyya P; Membrane vesicles isolated from cells of bacillus subtilis W23 accumulate manganese in the presence of an energy source . The artificial electron donor system ascorbate and phenazine methosulfate or reduced nicotinamide adenine dinucleotide and phenazine methosulfate can supply the energy for the uptake . D-Lactate in the presence or absence of phenazine methosulfate would not support manganese accumulation . Anaerobiosis, cyanide, m-chlorophenyl carbonylcyanide hydrozone, valinomycin, gramicidin, and p-hydroxy-mercuribenzoate inhibit the uptake . The inhibition by p-hydroxymercuribenzoate is prevented by excess dithiothreitol . Potassium fluoride or sodium arsenate has no effect on the uptake . The manganese transport system in the B . subtilis vesicles exhibits Michaelis-Menten kinetics with a Km of 13 muM and a Vmax of 1.7 nmol/min per mg (dry weight) of membranes . The uptake of manganese is specific and is not inhibited by 0.1 mM CaCL2 or Mgcl2. Biochim Biophys Acta, 1975 Jun 26, 393(2), 571 - 82 Hybrid alpha-amylases produced by the transformants of Bacillus subtilis . III . A possible mechanism of formation of hybird alpha-amylases; Yamane K et al.; Alpha-Amylases (NA64 and NA20) produced by the representative transformants Bacillus subtilis NA64 and NA20 were hybrid enzymes between the two parental alpha-amylases (NAT and MAR) produced by the DNA donor strain of Bacillus natto IAM 1212 and the DNA recipient strain of B . subtilis 6160, a derivative of B . subtilis 168 . In order to elucidate a possible mechanism of formation of the hybrid alpha-amylases, 14C-labeled alpha-amylase (SAC) produced by B . subtilis var . amylosarcchariticus, {3H}lysine- and {3H}arginine-labeled alpha-amylases (MAR, NA64, NA20, NAT and SAC), {3H}lysine-labeled alpha-amylase (SAC) and {3H}glucosamine-labeled alpha-amylase (NA64) were purified through ammonium sulfate precipitation, carboxy-methylcellulose and DEAE-Sephadex A-50 column chromatography and immunoprecipitation with rabbit antiserum against alpha-amylase (SAC) . Peptide compositions of the tryptic digests from the labeled alpha-amylases were analyzed by double-label AG 50W-X2 column chromatography . On the other hand, amino- and carboxy-terminal amino acid residues of unlabeled alpha-amylases (MAR, NA64, NA20 and NAT) were analyzed . Based on these results, the possibility of DNA recombination events in the alpha-amylase structure gene and on the previous results, we attempted to estimate possible peptide arrangements for the four alpha-amylases (MAR, NA64, NA20 and NAT) and possible recombination regions to form the hybrid enzymes introduced by the DNA-mediated transformation of B . subtilis 6160. J Biol Chem, 1975 Jun 25, 250(12), 4530 - 41 RNA polymerase from phage SP01-infected and uninfected Bacillus subtilis; Duffy JJ et al.; A purification procedure for RNA polymerase from uninfected and phage SP01-infected Bacillus subtilis is presented . The RNA polymerase purified from B . subtilis 10 min after infection with wild type phage SP01 is resolved into two major fractions (B, C) and one minor fraction (A) by calf thymus DNA-cellulose chromatography . Fraction C is indistinguishable from RNA polymerase from uninfected cells with respect to transcription specificity (both before and after phosphocellulose chromatography) . Fraction B yields, on subsequent phosphocellulose chromatography, an enzyme (B-P) whose properties distinguish it from the host RNA polymerase . Enzyme B-P preferentially transcribes SP01 DNA and selectively forms rapidly initiating complexes with SP01 DNA but not with heterologous DNA . The SP01 RNA synthesized by Enzyme B-P includes, as previously reported, a large proportion of asymmetrical middle viral RNA . Host RNA polymerase holoenzyme synthesizes asymmetrical early viral RNA, while host core polymerase synthesizes symmetrical RNA that is complementary to early, middle, and late in vivo viral RNA and contains a preponderance of antimessenger . The subunit composition of Enzyme B-P is identical to host core polymerase with respect to the beta,beta', and alpha subunits and two additional components of mr equals 9,500 and 11,000 that we observe in all preparations of RNA polymerase . In addition, Enzyme B-P has two subunits of mr equals 13,000 and 28,000, which are synthesized after phage infection . On heterologous template, Enzyme B-P and host core polymerase have comparable activities . On these templates, addition of host initiation factor, sigma, restores full activity to Enzyme B-P as well as to host core polymerase . Sigma also modifies the activity of Enzyme B-P on SP01 DNA, restoring some asymmetrical early RNA transcription while retaining some asymmetrical middle RNA transcription. J Biol Chem, 1975 Jun 25, 250(12), 4462 - 9 Anthranilate synthase from Bacillus subtilis . The role of a reduced subunit X in aggregate formation and amidotransferase activity; Holmes WM et al.; With respect to its sulfhydryl groups, subunit X can exist in at least two forms, oxidized (Xox) and reduced (Xre) . The importance of the Xre form for the formation of an EX complex and for amidotransferase activity has been examined . Subunit Xre is rapidly inactivated by p-chloromercuribenzoate and bromopyruvate, whereas subunit Xox, which is not catalytically functional in amidotransferase activity, is not affected . The glutamine analogue 6-diazo-5-oxo-L-norleucine (DON) has no effect on Xre alone but rapidly inactivates the EXre complex . DON-inactivated subunit X cannot be reactivated by 2-mercaptoethanol but can be readily displaced from subunit E by free subunit Xre . The integrity of the EXre complex is maintained following gel filtration on Sephadex G-100 in the presence of glutamine and 2-mercaptoethanol, thus the binding of glutamine to the complex does not require the binding of other substrates . Subunit Xox, however, does not aggregate with subunit E since no EXox complex is found following gel filtration on Sephadex G-100 in the presence of glutamine and in the absence of 2-mercaptoethanol . Thus, a reduced sulfhydryl group(s) is not only essential for amidotransferase activity but also for the formation of the aggregate as well . The following model is proposed to explain these results . Free subunit Xre does not bind DON or glutamine to the catalytically functional sulfhydryl group . Upon aggregation with subunit E, however, the glutamine or DON binds to the glutamine catalytic site on subunit Xre and amidotransfer or alkylation occurs . An EX complex which has been alkylated by DON can be readily dissociated and it is suggested that following catalysis the EX complex may also dissociate. Tijdschr Diergeneeskd, 1975 Jun 15, 100(12), 662 - 8 {False-positive results obtained on examining slaughtered animals for the presence of antibiotic residues (author's transl)}; Nouws JF; As part of the examination of emergency-slaughtered animals for the presence of antibiotic residues, studies were done to see whether false-positive results would be obtained when the Sarcina lutea kidney test and Bacillus subtilis BGA test were performed . When the S . lutea kidney test was positive in cattle, calves and swine, penicillin was invariably found to be present in those animals, the histories of which showed that they had not been given antibiotics . A syringe and an injected fluid containing penicillin residues are regarded as possible causes of these positive results . When the S . lutea kidney test was performed in horses which had been in a state of stress prior to slaughter, false-positive results were occasionally observed . When the German B . subtilis BGA test was used, a large number of false-positive results were recorded, among others in equine kidneys . This shows that the use of this test in examining the kidney is a highly controversial matter . A number of drugs used in veterinary medicine may have an inhibitory effect on the growth of S . lutea and B . subtilis BGA test organisms in vitro . When these drugs were used therapeutically (in vivo), the result of the S . lutea kidney test was not positive in any of the cases studied . On the other hand, false-positive results were obtained when the B . subtilis BGA renal medullary test was performed in those animals in which lidocaine and calcium borogluconate had also been injected. J Gen Microbiol, 1975 Jun, 88(2), 289 - 94 Properties of some norvaline-resistant mutants of Bacillus subtilis; Holtzclaw WD et al.; DL-Norvaline inhibits growth of wild-type Bacillus subtilis . A number of mutants resistant to growth inhibition by this analogue were isolated and studied . Cross-feeding experiments and paper chromatography of culture supernatants indicated that the mutants excreted leucine and possibly valine and glutamate . Enzymic analysis indicated that the mutants were derepressed for acetohydroxy-acid synthetase and alpha-isopropylmalate synthetase; however, no derepression of threonine deaminase, dihydroxyacid dehydrase or transaminase B was observed. J Bacteriol, 1975 Jun, 122(3), 987 - 93 Transformation of Bacillus subtilis: transforming ability of deoxyribonucleic acid in lysates of L-forms or protoplasts; Bettinger GE et al.; The transformation of Bacillus subtilis by homologous deoxyribonucleic acid (DNA) made available by gently lysing a stable L-form or protoplast suspension was 3 to 10-fold more efficient than DNA isolated by conventional procedures . This increased transformation was not influenced by digestion with pronase, trypsin, or ribonuclease . Preincubation of isolated DNA with L-form lysates did not increase the transformation efficiency above that achieved with untreated, isolated DNA . In addition to displaying a higher efficiency of transformation, the DNA found in these gently prepared lysates was also able to co-transform heretofore unlinked markers at frequencies in excess of those found by congression . Comparison of the frequency of multiple marker transformations to single marker events as a function of DNA dilution conclusively proves that these markers originated from the same continuous strand of DNA. J Bacteriol, 1975 Jun, 122(3), 886 - 98 Evidence for the Translocation of a Chromosome Sement in Bacillus subtilis Strains Carrying the trpE26 Mutation; Trowsdale J et al.; The replication order of markers was studied in Bacillus subtilis strains bearing the trpE26 mutation by the use of the density transfer technique . An important difference in this order was observed in comparison with that of strain 168 T- . All markers tested of a chromosome segment extending from trpD to ilvA replicated early, after purB6 and before thr-5 . Two markers flanking this region, trpE8 and citK7, replicated late as usual . The results suggested that this segment was shifted in trpE26 strains to a region closer to the origin of replication . PBS-1-mediated transduction crosses corroborated this hypothesis and revealed the position of the translocated segment . (i) Linkage was demonstrated for markers in the segment (hisH2, tryA1, met B3, ilvA2) to thr-5 and ald; (ii) aroB2 and citK7 were found to be linked; and (iii) linkage of cysB3 to thr-5 was lost in trpE26 strains . These findings made it possible to account for the characteristics of the trpE26 mutation and to propose a model explaining the fact that all trpE26+ transformants or transductants are merodiploid . The model calls for fusion of two genetic elements: two independent chromosomes, or two arms of a replicating structure . The resulting chromosome bears a long tandem duplication . Most of the features of this system of merodiploid formation can be interpreted by use of this model: the segregation pattern of the diploids, the stabilization of the unstable clones, and the length of the duplicated region . A relatively stable diploid strain was also studied by the density transfer technique . The data show that it remained diploid for the region corresponding to the translocated segment and are in agreement with the structure predicted by the model. J Bacteriol, 1975 Jun, 122(3), 880 - 5 Facilitated transport of calcium by cells and subcellular membranes of Bacillus subtilis and Escherichia coli; Silver S et al.; The level of calcium in growing cells is lower than that in the growth medium . Non-energy-dependent uptake of 45-Ca by log-phase cells of Bacillus subtilis occurs under two conditions: at 0 C or in the presence of m-chlorophenyl carbonylcyanide hydrazone . Similar uptake, but quantitatively less, occurs with Escherichia coli cells under the same conditions . Membrane vesicles prepared from B . subtilis or E . coli accumulate 45-Ca by a process that does not depend on added energy sources and is not inhibited by the respiratory poison cyanide . The properties of calcium transport in all cases is consistent with carrier-mediated, facilitated transport with specificity Ca-2+ greater than Sr-2+ greater than Mn-2+ greater than Mg-2+ . Upon transfer of cells from 0 C to 20 C, pre-accumulated 45-Ca is released . Heat-killed cells do not accumulate 45-Ca and calcium is released by cells upon addition of toluene (under conditions that do not cause visible lysis) . These results suggest that the facilitated uptake of calcium may be utilizing a transport system that normally is responsible for the energy-dependent excretion of calcium from the cells. J Bacteriol, 1975 Jun, 122(3), 1387 - 90 Rifampin resistance mutation of Bacillus subtilis altering the electrophoretic mobility of the beta subunit of ribonucleic acid polymerase; Linn T et al.; The rifampin-resistance mutation of LS3,an asporogenous mutant of Bacillus subtilis 3610, leads to altered mobility of the beta subunit of ribonucleic acid polymerase in sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis . This finding argues that the rifampin-resistance mutation is located in the structural gene coding for the beta polypeptide. J Bacteriol, 1975 Jun, 122(3), 1109 - 16 Genetic mapping of sporulation operons in Bacillus subtilis using a thermosensitive sporulation mutant; Young M; A thermosensitive sporulation mutant was used to determine the order of sporulation operonsin the urs region of the Bacillus subtilis chromosome . Data from three-factor transformation crosses and three- and four-factor transduction crosses established the order metC-SPO-96(SpoII)-spo-85(SpoV)-spo-279(SpoII)-furA-ura-cysC-spo-NG1.67(SpoIII) . Previously, furA was thought to lie to the right of ura and cysC to the left (Dubnau, 1970; Young and Wilson, 1972). J Virol, 1975 Jun, 15(6), 1308 - 16 Bacteriophage SPO1-induced macromolecular synthesis in minicells of Bacillus subtilis; Reeve JN et al.; SPO1 bacteriophage injects its DNA into minicells produced by Bacillus subtilis CU403 divIVB1 . The injected DNA is partially degraded to small trichloracetic acid-precipitable material and trichloroacetic acid-soluble material . The injected DNA is not replicated; however, it serves as a template for RNA and protein synthesis . The RNA produced specifically hybridizes to SPO1 DNA, and the amount of RNA hybridized can be reduced by competition with RNA isolated at all stages of the phage cycle from infected nucleate cells of the B . subtilis CU403 divIVB1 . An unrelated phage, SPP1, also induces phage-specific RNA in infected minicells . Translation occurs in SPO1-infected minicells resulting in at least eight proteins which have been separated by gel electrophoresis, and two of these proteins have mobilities similar to proteins found only in infected B . subtilis CU403 divIVB1 nucleate cells . A large proportion of the polypeptide material synthesized in infected minicells is very small and heterogeneous in size. Bull Environ Contam Toxicol, 1975 Jun, 13(6), 689 - 91 Bacillus subtilis; a sensitive bioassay for patulin; Reiss J; The inhibition of the germination of Bacillus subtilis spores by patulin provides a sensitive and simple technique for the microbiological detection of this mycotoxin . The paper disc assay procedure is performed in standardized plastic plates containing agar and spores . Significant inhibition was obtained with as little as 1 microgram patulin. Proc Natl Acad Sci U S A, 1975 Jun, 72(6), 2366 - 70 Conversion of Bacillus subtilis RNA polymerase activity in vitro by a protein induced by phage SP01; Duffy JJ et al.; A protein fraction from B . subtilis infected with phage SP01 (fraction LGG) stimulates the activity of RNA polymerase (EC 2.7.7.6; nucleosidetriphosphate:RNA nucleotidyltransferase) core from uninfected bacteria . Fraction LGG contains a protein (P-28, molecular weight 28,000) that is labeled after phage infection and binds tightly to RNA polymerase core at a relatively high ionic strength . B . subtilis RNA polymerase core with bound P-28 has the transcription specificity of the previously purified, phage-modified B-P RNA polymerase; the latter contains two subunits, v-28 and v-13 (molecular weights 28,000 and 13,000, respectively) that are synthesized after phage infection . Both enzymes transcribe SP01 DNA preferentially and direct the asymmetric synthesis of viral middle RNA . P-28, like v-28, binds more tightly to B . subtilis RNA polymerase core than the B . subtilis initiation factor, sigma, at higher ionic strength . We propose that P-28 and v-28 are the same protein . P-28 and, by implication, v-28 suffice to endow the bacterial RNA polymerase core with a novel transcription specificity. Proc Natl Acad Sci U S A, 1975 Jun, 72(6), 2207 - 11 Electrophoretic separation of Bacillus subtilis genes; Harris-Warrick RM et al.; The cleavage of Bacillus subtilis DNA by EcoR1 restriction endonuclease produced segments which retain various degrees of genetic transforming activity . The active segments analyzed thus far, range in size from 23 to 3 kilobases and can be partially separated by agarose gel electrophoresis . Various markers can thus be enriched from 30- to 60-fold. Eur J Biochem, 1975 Jun, 54(2), 475 - 82 Synthesis of coenzymically active soluble and insoluble macromolecularized NAD+ derivatives; Zappelli P et al.; Alkylation at N-1 of the NAD+ adenine ring with 3,4-epoxybutanoic acid, followed by chemical reduction to the alkali-stable NADH form and alkaline Dimroth rearrangement, gave the NADH derivative alkylated at the exocyclic adenine amino group . Enzymic reoxidation of the latter derivative gave nicotinamide-6-(2-hydroxy-3-carboxypropylamino)purine dinucleotide, a functionalized NAD+ analogue carrying an omega-carboxyalkyl side-chain at the exocyclic adenine amino group . Carbodiimide coupling of the latter derivative to high-molecular-weight water-soluble (polyethyleneimine, polylysine) and insoluble (aminohexyl-Sepharose) polymers gave the corresponding macromolecularized NAD+ analogues . These derivatives have been shown to be enzymically reducible . The polyethyleneimine and polylysine analogues showed a substantial degree of efficiency relative to free NAD+ with rabbit muscle lactate dehydrogenase (60 and 25% respectively) but a lower one with yeast alcohol dehydrogenase and Bacillus subtilis alanine dehydrogenase (2-7%) . The polyethyleneimine derivative entrapped in cellulose triacetate fibres together with the lactate dehydrogenase was operationally stable during repetitive use. J Antibiot (Tokyo), 1975 Jun, 28(6), 448 - 52 On the effect of N-methyl-bis (3-mesyloxypropyl) amine hydroxychloride on Bacillus subtilis cells; Shimi IR et al.; N-Methyl-bis (3-mesyloxypropyl)amine hydrochloride is now in use as an antitumer drug . In view of its activity against some bacteria the present work was conducted to study its mode of action of Bacillus subtilis . The compound was found to induce irreversible damage to bacterial DNA whereas its effect on RNA was temporary and depending on maintenance of effective concentrations of the compound. Antibiotiki, 1975 Jun, 20(6), 499 - 502 {Determination of the biological activity of heliomycin by a method of diffusion in agar}; Kochetkova GV et al.; A biological method for determination of geliomycin activity using agar diffusion is described . A nutrient medium containing Hottinger broth up to 35 mg per cent of amine nitrogen, 1.5 percent of agar-agar, tap water, pH 7.8 to 8.0 was used . Bacillus subtilis ATCC 6633 served as the test-culture . Geliomycin was dissolved in 0.1 N sodium hydroxide solution. Prikl Biokhim Mikrobiol, 1975 May-Jun, 11(3), 397 - 405 {Inosine synthesis by Bacillus subtilis mutants and their development in synthetic media}; Balitskaya RM et al.; The inosine synthesis by Bacillus subtilis mutants selected by the Institute of Genetics 265 and 21 and their development on the mineral medium were studied . The mutants behaved differently towards the sources and concentrations of carbon, nitrogen, phosphorus and magnesium as well as towards additions of tyrosine, histidine, adenine and uracyl . Optimal concentrations of the components of the mineral medium for the inosine synthesis by the Bac . subtilis mutants were determined . Under favourable conditions the mutants synthesized 8 to 10 mg/ml inosine for 120 hours. Prikl Biokhim Mikrobiol, 1975 May-Jun, 11(3), 391 - 6 {Effect of carbon and nitrogen sources and complex B vitamins on the synthesis of alkaline protease by different strains of Bacillus mesentericus and Bacillus subtilis}; Emtseva TV; The effect of different sources of carbon, nitrogen, amino acids and vitamins on the synthesis of alkaline proteases by the stock and mutant strains of Bacillus mesentericus and by the natural strain of Bacillus subtilis-12 has been investigated . The maximum synthesis of alkaline protease has been obtained in the media containing starch or its hydrolysates--dextrine and maltose as the carbon source . Ammonium phosphate and casein as the nitrogen source prove to be optimal for Bac . mesentericus and Bac . subtilis, respectively . Complex B vitamins added to the nutrient medium accelerate the enzyme synthesis 2.5-4-fold. Prikl Biokhim Mikrobiol, 1975 May-Jun, 11(3), 362 - 6 {Influence of clay minerals on the oxidative activity of the caprolactam destructors Bacillus subtilis 6 and 21}; Rotmistrov MN et al.; By the Warburg manometric method the respiratory activity of the caprolactam destructors--Bacillus subtilis strains 6 and 21 was measured . The bacteria grown on the synthetic nutrient medium with caprolactam as the sole source of carbon and nitrogen oxidized that substrate more intensively than the cells grown on meat-peptone agar . The activity of caprolactam oxidation by the cultures showed strain differences . Clay minerals--montmorillonit, palygorskit, bentonit and vermiculit--stimulated the glucose and caprolactam oxidation by the above bacteria . The highest stimulating effect was produced by montmorillonit. J Gen Microbiol, 1975 May, 88(1), 115 - 22 Thymineless death in Bacillus subtilis; Buick RW et al.; A study has been made of the intracellular changes occurring during thymineless death in Bacillus subtilis 2337 . The kinetics of death were paralleled by the rate of breakdown of DNA . During thymineless death single-strand breaks accumulated within DNA, breakdown of approximately 13% of the DNA to acid-soluble material occurred, deoxyribonuclease levels rose sharply, and yet double-strand breaks did not occur in the DNA . On restoration of thymine, however, double-strand breaks accumulated, though this could be prevented by the specific inhibition of DNA preplication on restoration of thymine . The results probably indicate that during thymineless death, single-strand gaps accumulate within the DNA of cells . On restoration of thymine both DNA replicaion and repair of gaps are simultaneously initiated, and when a replication fork reaches a gap before it is repaired, double-strand breakage of the DNA occurs . The possible relevance of these events to the lethality of the cells is discussed. J Virol, 1975 May, 15(5), 1286 - 8 DNA strand specificity of transcripts produced in vivo and in vitro by RNA polymerase from SP82-infected Bacillus subtilis; Lawrie JM; Phage-specific RNA synthesized early in the infection of Bacillus subtilis with SP82 hybridizes to both heavy (H) and light (L) strands of SP82 DNA nearly equally . Phage RNA synthesized during the middle of the infection hybridizes preferentially to the H strand . The ratio of H/L strand binding of RNAs synthesized in vitro by RNA polymerases isolated from uninfected and infected cells resembles the ratios of early and middle phage RNA classes, respectively . This supports the conclusion that a modified RNA polymerase is required for the transcription of middle RNA classes. J Virol, 1975 May, 15(5), 1073 - 80 Changes in DNase activities in Bacillus subtilis infected with bacteriophage PBS 1; Tomita F; DNase activities in Bacillus subtilis were fractionated by chromatography on hydroxylapatite . One of the fractions hydrolyzed uracil-containing phage DNA but had no activity on host DNA . This activity on native phage DNA disappeared soon after phage infection, whereas DNase activities on bacterial DNA remained at the same level during phage development . An inhibitor of protein nature was induced by phage infection and this inhibitor was shown to be responsible for the disappearance of the DNase activity on phage DNA . Bacterial DNA in infected cells might be fragmented but is not degraded to acid-soluble oligonucleotides. J Bacteriol, 1975 May, 122(2), 610 - 22 Fate of heterologous deoxyribonucleic acid in Bacillus subtilis; Piechowska M et al.; CsCl density gradient fractionation of cell lysates was employed to follow the fate of Escherichia coli, phage T6, and non-glucosylated phage T6 deoxyribonucleic acid (DNA) after uptake by competent cells of Bacillus subtilis 168 thy minus trp minus . Shortly after uptake, most of the radioactive Escherichia coli or non-glucosylated T6 DNA was found in the denatured form; the remainder of the label was associated with recipient DNA . Incubation of the cells after DNA uptake led to the disappearance of denatured donor DNA and to an increase in the amount of donor label associated with recipient DNA . These findings are analogous to those previously reported with homologous DNA . By contrast, T6 DNA, which is poorly taken up, appeared in the native form shortly after uptake and was degraded on subsequent incubation . The nature of the heterologous DNA fragments associated with recipient DNA was investigated with Escherichia coli 2-H and 3-H-labeled DNA . Association of radioactivity with recipient DNA decreased to one-fourth in the presence of excess thymidine; residual radioactivity could not be separated from recipient DNA by shearing (sonic oscillation) and/or denaturation, but was reduced by one-half in the presence of a DNA replication inhibitor . Residual radioactivity associated with donor DNA under these conditions was about 5% of that originally taken up . Excess thymidine, but not the DNA replication inhibitor, also decreased association of homologous DNA label with recipient DNA; but, even in the presence of both of these, the decrease amounted to only 60% . It is concluded that most, or all, of the Escherichia coli DNA label taken up is associated with recipient DNA in the form of mononucleotides via DNA replication. J Bacteriol, 1975 May, 122(2), 526 - 31 Alteration of tyrosine isoaccepting transfer ribonucleic acid species in wild-type and asporogenous strains of Bacillus subtilis; McMillian RA et al.; The relative amounts of two isoacceping species of tyrosine transfer ribonucleic acid, tRNATyrI and tRNATyrII, determined from reversed phase 5 profiles of tyrosyl-tRNA, prepared from Bacillus subtilis strain W168, were growth phase and medium dependent . The growth phase-dependent alterations in the relative amounts of tRNATyr species were also demonstrated in 11 asporogenous strains of B . subtilis . The proportion of tRNA-Tyr species and the extent of the alteration in their relative amounts during the transition from the exponential to the stationary phase of growth of these strains was not directly correlated with the formation of spores by strain W168 grown in various media or the stage at which the asporogenous strains are blocked in the process of sporulation. J Biochem (Tokyo), 1975 May, 77(5), 957 - 63 The pH jump study of enzyme proteins . I . Liquefying alpha-amylase from Bacillus subtilis; Hiromi K et al.; Rapid conformational changes due to pH jump were studied kinetically at 25 degrees mainly by the stopped-flow method using liquefying alpha-amylase from Bacillus subtilis {EC 3.2.1-.1, liquefying} . First, the conformational change due to a pH jump produced by mixing with alkali was monitored as a function of time at 245 nm through the ionization of phenolic hydroxyl groups of tyrosine residues which were originally buried and finally become exposed due to the pH jump . Three distinct phases of conformational change were clearly recognized by this method by varying the final pH values . Each phase involved the exposure of an essentially definite number of tyrosine residues, whose rate constant was crucially dependent on pH . Second, these phases of conformational change were subjected to examination in terms of the optical rotation change at 411 nm and the reversibility upon reverse pH jump with respect to conformational reconstitution, as observed through the protonation ofphenolic hydroxyl groups of ionized tyrosine residues and the enzyme activity . The first phase, which occurs above pH 12.5, involves no change in the optical rotation and is reversible as observed by the above two monitoring methods . In contrast, the other two phases, which are observed above pH 12.7, are accompanied by an optical rotation change and no appreciable reversibility was detected by these methods. Can J Microbiol, 1975 May, 21(5), 639 - 47 Relationship between the DNA content and mesosome number in cells of Bacillus; Johnston GC; In cells of Bacillus there is evidence that deoxyribonucleic acid forms an association with some membranous structure within the cell, possibly mesosomes . Cells of varieties of Bacillus cereus and Bacillus subtilis were examined to see if any quantitative relationship existed between the numbers of mesosomes and DNA content . No direct relationship could be domonstrated . However, cells of Bacillus cereus var . alesti A(-) maintained a characteristic and constant DNA content and number of mesosomes regardless of growth conditions . During sporulation, a variant of A(-), termed A(-)3, SEQUESTERS ITS DNA at both ends of the cell, leaving a small amount of DNA but no mesosomes in the center compartment . Since this center compartment is capableof growth and division upon replacement in fresh medium (rejuventation) it was examinedfor mesosome content as DNA synthesis and division were initiated . In most cells, acentral mesosome was formed at the site of cell septum formation; however, the presenceof a mesosome was not an absolute prerequisite for cell division . We propose that atthe onset of cell growth, mesosomes primarily function in the process of cell septum formation . As growth and division proceed, mesosomes are produced in characteristicnumbers and may act as the site of DNA synthesis and (or) segregation. J Biol Chem, 1975 Apr 25, 250(8), 2813 - 22 Purification and characterization of an inhibitor of phospholipase A1 in Bacillus subtilis; Krag SS et al.; The protoplasts of a mutant of Bacillus subtilis 168 (B . subtilis CMK33) are osmotically fragile when compared to protoplasts of the parent organism and contain an active, membrane-associated phospholipase A1 . A protein found in the parent organism specifically inhibits the phospholipase A1 (Kent, C., and Lennarz, W.J . (1972) Proc . Nat . Acad . Sci . U.S.A . 69, 2793-2797) . The inhibitor exists in both a soluble and particulate form . The soluble inhibitor is not found in the cytoplasm, but rather in a "periplasmic" fraction released from the cell during incubation with lysozyme . The soluble inhibitor has been purified to homogeneity by diethylaminoethyl-cellulose and hydroxylapatite chromatography . Its molecular weight is 28,000 to 32,000 as determined by gel filtration chromatography and 36,000 to 37,000 as determined by sodium dodecyl sulfate-urea gel electrophoresis . The inhibitor appears to inactivate the membrane bound phospholipase A1 by an enzymatic process that is dependent on time and protein concentration . Binding of the inhbitor to the membrane-associated phospholipase cannot be detected . When purified inhibitor is added to cells of B . subtilis CMK33 during treatment with lysozyme, the osmotic stability of the resultant protoplasts is similar to that of protoplasts of the wild type of organism. Biochim Biophys Acta, 1975 Apr 2, 383(4), 379 - 87 Depression by NAD of x-ray-induced repair-type DNA synthesis in toluene-treated Bacillus subtilis; Billen D et al.; NAD prevents a DNA repair-type synthesis that is dependent on polymerase I in toluene-treated, X-irradiated Bacillus subtilis . In unirradiated preparations, NAD had little effect on an ATP-dependent, semiconservative synthesis but partially inhibited a repair-type synthesis . In a mutant lacking polymerase I (polA1-), the presence of NAD did not affect dTTP utilization in DNA synthesis . Nicotinamide mononucleotide (NMN) partially reverses the NAD inhibition of repair-type DNA synthesis . NADP and FAD were ineffective as substitutes for NAD . Since NAD is the cofactor for polynucleotide ligase in Bacillus subtilis and NMN is known to discharge AMP from the active AMP ligase complex, it is proposed that activation of DNA ligase reduces dTMP incorporation by reducing sites for, or limiting DNA polymerase I action. Biochem J, 1975 Apr, 147(1), 187 - 9 The teichuronic acid of cell walls of Bacillus subtilis W23 grown in a chemostat under phosphate limitation; Wright J et al.; Cell walls of Bacillus subtilis W23 contain teichuronic acid when grown in a chemostat under phosphate limitation at a low dilution rate, but teichoic acid at a higher dilution rate . The teichuronic acid was purified and shown to be a polymer of glucuronic acid and N-acetylgalactosamine. J Antibiot (Tokyo), 1975 Apr, 28(4), 266 - 73 Metabolic products of microorganisms 142 . A new antibiotic derinamycin, inhibitor of DNA and RNA synthesis; Uchida K et al.; Derinamycin was isolated from the myecelium of Streptomyces venezuelae Tu 1102 and its molecular formula was tentatively assigned as C51H93NO23 . The antibiotic inhibits the growth of fungi, gram-positive bacteria and certain gram-negative bacteria but is less acitve against yeasts . A study of derinamycin action on the macromolecular synthesis of intact Bacillus subtilis revealed that the antibiotic suppressed DNA and RNA syntheses but that protein synthesis was less affected . Derinamycin exerted no selective inhibition between DNA and RNA syntheses in the double-isotope experiment used to assess the relative effects of the antibiotic. Br J Radiol, 1975 Apr, 48(568), 306 - 11 Absence of mutation following ultrasonic treatment of Bacillus subtilis cells and transforming deoxyribonucleic acid; Combes RD; Possible mutagenic effects of ultrasound at medical dosages have been assessed using genetic systems of Bacillus subtilis . The induction of mutations, after treatment of cells and of extracted transforming DNA with ultrasound has been tested . High-frequency (2 MHz diagnostic regime and higher intensities) ultrasound was unable to increase significantly the spontaneous frequency of back-mutation of an auxotrophic strain . Moreover, high-frequency treatments (1.5 MHz diagnostic and therapeutic regimes) were incapable of producing detectable levels of mutagenic lesions after in vitro irradiation of transforming DNA . Slight decreases in transforming activity of the treated DNA were apparent while the degree of linkage between two contiguous markers was unaffected . It is concluded that the ultrasound treatments employed under the conditions pertaining do not result in production of detectable mutagenic effects in cells or in vitro treated DNA . Before extrapolating such results to the human hazard situation, it is suggested that tests using genetic systems of higher organisms should be carried out. Proc Natl Acad Sci U S A, 1975 Apr, 72(4), 1589 - 93 Highly asymmetric transcription by RNA polymerase containing phage-SP01-induced polypeptides and a new host protein; Pero J et al.; An RNA polymerase (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) has been purified from phage-SP01-infected Bacillus subtilis that copies RNA almost exclusively from the heavy strand of native SP01 DNA, the DNA strand from which "middle" and "late" classes of RNA are copied in vivo . Hybridization-competition established that this RNA polymerase termed enzyme A, preferentially synthesizes middle RNA in vitro . Enzyme A contains beta',beta, alpha, and two newly identified host polypeptides, variation of (21,500 daltons) and omega (11,000 daltons) . All of these polypeptides are associated with highly purified RNA polymerase from uninfected bacteria . In addition, enzyme A contains phage-induced subunits of 26,000, 24,000, and 13,500 daltons . Enzyme A lacks sigma polypeptide, and strand-selective transcription by this enzyme is resistant to anti-sigma antibody . A reconstitution experiment strongly suggests that the host variation of protein is required in addition to a phage-induced subunit(s) (or an unidentified phage-induced modification) for strand-selective transcription of SP01 middle genes in vitro. J Bacteriol, 1975 Apr, 122(1), 25 - 33 Different nuclease activities in competent and noncompetent Bacillus subtilis; Joenje H et al.; Competent and noncompetent cells of Bacillus subtilis were separated on the basis of their different buoyant densities . The two types of cells were compared with respect to their interactions with exogenous deoxyribonucleic acid(DNA) . After exposure of DNA to the cells, the unadsorbed fraction of DNA molecules was examined . Both types of cells decreased the biological activity of this DNA, the inactiviation exerted by noncompetent cells being more severe than that exerted by competent cells . Sedimentation analysis of the inactivated DNA revealed that fragments of DNA are produced, owing mainly to the introduction of double-strand scissions . In addition to this fragmentation, the competent bacteria extensively digested the DNA exonucleolytically . This type of breakdown was specifically related to the competent state rather than to the state of low density . The exonucleolytic activity is, in all probability, associated with the cell envelope, because most of the activity is released into the medium when the cells are converted to protoplasts . At 37 C the competence-specific exonucleolytic breakdown started 2 to 3 min after the binding of DNA to the cells . In unfractionated cultures, breakdown may proceed until 70% of the total amount of DNA added has been made acid soluble . Nontransforming Escherichia coli DNA was also subject to exonucleolytic degradation; it seems unlikely,therefore, that this type of breakdown occurs as a consequence of recombination . Since ethylenediaminetetraacetate blocked both transformation by native DNA and the exonucleolytic breakdown of bound DNA, we suggest that the breakdown of DNA by competent cells fulfills an essential function in genetic transformation of B . subtilis. J Bacteriol, 1975 Apr, 122(1), 224 - 34 Regulation of the dicarboxylic acid part of the citric acid cycle in Bacillus subtilis; Ohne M; The regulation of alpha-ketogluterate dehydrogenase, succinate dehydrogenase, fumarase, malate dehydrogenase, and malic enzyme has been studied in Bacillus subitilis . The levels of these enzymes increase rapidly during late exponential phase in a complex medium and are maximal 1 to 2 h after the onset of sporulation . Regulation of enzyme synthesis has been studied in the wild type and different citric acid cycle mutants by adding various metabolites to the growth medium . Alpha-ketoglutarate dehydrogenase is induced by glutamate or alpha-ketoglutarate; succinate dehydrogenase is repressed by malate; and fumarase and malic enzyme are induced by fumarate and malate, respectively . The addition of glucose leads to repression of the citric acid cycle enzymes whereas the level of malic enzyme is unaffected . Studies on the control of enzyme activities in vitro have shown that alpha-ketoglutarate dehydrogenase and succinate dehydrogenase are inhibited by oxalacetate . Enzyme activities are also influenced by the energy level, expressed as the energy charge of the adenylate pool . Isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, and malic enzyme are inhibited at high energy charge values, whereas malate dehydrogenase is inhibited at low energy charge . A survey of the regulation of the citric acid cycle in B.subtilis, based on the present work and previously reported results, is presented and discussed. J Bacteriol, 1975 Apr, 122(1), 152 - 8 Distribution of teichoic acid in the cell wall of Bacillus subtilis; Doyle RJ et al.; Hydrolysis of the cell wall of Bacillus subtilis 168 by autolysins or lysozyme resulted in the exposure of glucosylated teichoic acid molecules as evidenced by increased precipitation of {14C} concanavalin A . The number of concanavalin A-reactive sites increased significantly after only limited enzymatic digestion of the walls . Quantitative analyses of {14C} concanavalin A-treated wall or wall hydrolysate complexes indicate that approximately one-half of the teichoic acid molecules are surface-exposed, whereas the remainder are probably embedded within the peptidoglycan matrix . Treatment of the cell walls with sodium dodecyl sulfate or Triton X-100 did not result in new concanavalin A-reactive sites . Partial autolysis diminished the ability of the cell walls to adsorb bacteriophage phi25 . Fluorescein-labeled concanavalin A bound intensely over the entire surface of growing B . subtilis 168 cells, suggesting that teichoic acid molecules are located on the total solvent-exposed surface area of the bacteria. Can J Microbiol, 1975 Apr, 21(4), 527 - 36 Regulation of C4-dicarboxylic acid transport in Bacillus subtilis; Ghei OK et al.; Various mutant strains of Bacillus subtilis were used to study the induction and regulation of the transport of tricarboxylic acid cycle C4-dicarboxylates . L-Malate was the only physiological inducer and bromosuccinate was a gratuitous inducer of dicarboxylic acid transport in a succinic dehydrogenase deficient mutant . Several mutants were isolated with alterations in the ability to transport dicarboxylic acids . One of these (WK6) was defective in the ability to take up succinate when grown on glutamate minimal medium, whereas another (WK1) was inducible to high levels by extremely low levels of L-malate . Alpha-Ketoglutarate dehydrogenase mutants were unable to take up dicarboxylates because of repression of transport by glutamate and (or) alpha-ketoglutarate . A mutation which resulted in increased levels of alpha-ketoglutarate dehydrogenase partially overcame this inhibition . Glutamate did not prevent the induction of dicarboxylic acid transport by L-malate in succinic dehydrogenase mutants but was markedly inhibitory in alpha-ketoglutarate dehydrogenase mutants. J Virol, 1975 Apr, 15(4), 820 - 7 Viral mutation affecting bacteriophage phi 1 development in Bacillus subtilis 168; Rettenmier CW et al.; Bacillus subtilis bacteriophage phi 1m, a host-range variant, was isolated after mutagenesis of virulent bacteriophage phi 1 . Unlike its wild-type antecedent, phi 1m could not form plaques on lawns of B subtilis 168 at 37 C, although it adsorbed to, penetrated, and killed this bacterium . Experiments conducted in liquid medium at 37 C showed that B . subtilis 168 cells allowed reduced levels of phi 1m development at low multiplicities of infection, whereas high multiplicity infections of this strain by the phage were abortive . Certain mutants, derived originally from B . subtilis 168, were observed to be permissive for phi 1m at 37 C; moreover, their permissive phenotype could be duplicated by growing wild-type B . subtilis 168 cells at temperatures above 47 C . Studies on phi 1m and host nucleic acid synthesis under nonpermissive conditions demonstrated that transciption and DNA synthesis proceeded up to 20 min after infection, after which time there was a cessation of all nucleic acid production . These observations are discussed with respect to other abortive bacteriophage infections in B . subtilis. J Biol Chem, 1975 Mar 10, 250(5), 1676 - 82 Bacillus subtilis N-acetylmuramic acid L-alanine amidase; Herbold DR et al.; The N-acetymuramic acid L-alanine amidase from Bacillus subtilis (ATCC 6051) has been purified to homogeneity . It is a monomeric protein of molecular weight 50,000 . The enzyme has a high affinity for homologous cell walls and once attached to a cell wall will hydrolyze the wall completely before initiating the hydrolysis of a new cell wall . The affinity of the enzyme for cell walls devoid of teichoic acid or for cell walls of Bacillus megaterium is much lower than that for B . subtilis cell walls . A second homogenous protein has been isolated from B . subtilis which specifically combines with the amidase in a 1:1 molar ratio and stimulates enzyme activity . This modifier protein has no intrinsic cell wall lytic activity . The binding of enzyme and modifier protein has a dissociation constant of 8.5 times 10-9 M in 0.1 M LiCl, pH 8.0, but the two proteins can be completely dissociated in 3 M LiCl at pH 8.0. Proc Natl Acad Sci U S A, 1975 Mar, 72(3), 14 - 8 Versatile properties of a nonsaturable, homogeneous transport system in Bacilus subtilis: genetic, kinetic, and affinity labeling studies; Glover GI et al.; The mutliphasic kinetics that characterize the transport of many amino acids into Bacillus subtilis suggests a priori at least two mechanisms: (i) a tilis suggests a prior at least two independent transport processes, or (ii) a single, homogeneous system that might involve a negative cooperative mechanism . The highly specific transport system for L-tyrosine and L-phenylalanine in B . subtilis was studied as a case in point . The possible presence of a mixed system of independent transport systems was negated by the rentention of multiphasic kinetics of transport in two types of permease mutants . Furthermore, evaluation of kinetic data obtained during transport under various uptake conditions of pH and temperature, or in the presence of metabolic inhibitors, did not reveal the heterogeneity expected of mechanism (i) . These data, taken together with characteristics of substrate specificity and affinity labeling, provide substantial support for a negative cooperative mechanism for L-tyrosine and L-phenylalanine transport. Carbohydr Res, 1975 Mar, 40(1), 41 - 52 Extraction and purification of lipoteichoic acids from Gram-positive bacteria; Coley J et al.; Hot and cold, 80% aqueous phenol extraction procedures together with an aqueous extraction technique have been evaluated for the isolation of lipoteichoic acids from the cytoplasmic membrane of Gram-positive bacteria . Lipoteichoic acids of Staphlococcus aureus H, Micrococcus 2102, Baccillus subtilis 168, and Bacillus subtilis W-23 were examined as each of them emphasises a different problem of contamination . The purity of the lipoteichoic acids with respect to cell-wall material, nucleic acid, and protein is discussed together with the criteria of purity which enables critical structural analysis of lipoteichoic acids to be carried out. J Bacteriol, 1975 Mar, 121(3), 950 - 8 New transfer ribonucleic acid species during sporulation of Bacillus subtilis; Jeng YH et al.; The transfer ribonucleic acid (tRNA) populations from log-phase cells, sporulating cells (stage III), and dormant spores were compared by tRNA-deoxyribonucleic acid hybridization techniques . New tRNA species not found in log-phase cells were observed in stage III cells . Some of the tRNA made during sporulation were also present in dormant spores . Although the role and function of these new tRNA species cannot be ascribed directly to the sporulation process, their presence indicates that new tRNA genes can be transcribed during sporulation and suggests that translational control may be exerted during sporulation by tRNA. J Bacteriol, 1975 Mar, 121(3), 835 - 47 Bidirectional chromosome replication in Bacillus subtilis 168; Harford N; Density transfer analysis of deoxyribonucleic acid from Bacillus subtilis 168 thy spores germinating in 5-bromouracil medium shows the order of replication of genetic markers to be: purA16, cysA14, sacA, ctrA, (narB, arol), dal, (hisA1, purB6), (tre-12, thr-5), (argA, aroG, argC4), (metC, leu-8, pheA), (ura-1, aroD), lys-1, (trpC, metB, ilvA, citB, citK, gltA) . The precise order of transfer of markers within parentheses could not be determined in these experiments . Taken together with new PBS1 transduction data presented here and in the accompanying paper of J . Lepesant-Kejzlarova, J.-A . Lepesant, J . Walle, A . Billaut, and R . Dedonder (1975), the results can be resolved in terms of a symmetric, fully bidirectional mode of chromosome replication with a replication origin close to the purA16 marker and a terminus in the region of the gltA, citK loci, diametrically opposed to the origin . A new genetic map of the B . subtilis 168 chromosome is presented. J Bacteriol, 1975 Mar, 121(3), 823 - 34 Revision of the linkage map of Bacillus subtilis 168: indications for circularity of the chromosome; Lepesant-Kejzlarova J et al.; A revision of the linkage map of the Bacillus subtilis 168 chromosome has been undertaken with the use of the generalized transducing phage PBS1 . The mapping of four new markers (narB1, mtlB1, aroI906, and tre-12) has allowed a determination of the relative orientation of the purB-dal segment and its linkage with the lin markers . The chromosomal segment comprised between the sacQ36 and gtaA12 markers has been linked with the narA1, ctrA1, and sacA321 markers . The recA1 marker has been mapped relative to the thyA and citB17 markers . Indications of linkage have been found between the tre-12 and catA markers and the aroG932 and sacQ36 markers . According to these results, a circular genetic map of the chromosome of B . subtilis 168 is presented . Taken together, the transduction data and the order of marker replication determined by Harford in the accompanying paper support strongly the hypothesis of a symmetrical and fully bidirectional mode of replication for the B . subtilis 168 chromosome. J Bacteriol, 1975 Mar, 121(3), 771 - 6 Interactions between exogenous deoxyribonucleic acid and membrane vesicles isolated from competent and noncompetent Bacillus subtilis; Joenje H et al.; Competent cultures of Bacillus subtilis 168 have been fractionated into a high-competent and a low-competent fraction by a large-scale separation technique . Membrane vesicles isolated from both cell fractions are equally active in the transport of L-glutamate . Both membrane vesicle preparations seem to have similar endo- and exonuclease activities . Also, both preparations are capable of binding deoxyribonucleic acid . However, especially at low deoxyribonucleic acid concentrations (1 mug or less per ml), vesicles obtained from competent cells bind significantly more deoxyribonucleic acid (up to sixfold) than vesicles from noncompetent cells. J Bacteriol, 1975 Mar, 121(3), 1189 - 99 Nuclear and cell division in Bacillus subtilis: dormant nucleoids in stationary-phase cells and their activation; Van Iterson W et al.; The morphology of nucleoids and mesosomes of Bacillus subtilis in stationary-and lag-phase cultures was studied by making three-dimensional cell reconstructions in plastic of electron micrographs of serial sections . In cells from stationary cultures, the dormant nucleoids are frequently, but not always, spherical and the mesosomes are small and compact . It is suggested that the spherical nucleoids represent the resting stage in which replication and segregation have been completed . In cells from lag-phase cultures, the compact mesosomes develop into an elaborate system of tubes and wider sacs which become wrapped around the elongating nucleoids and invade the nucleoplasm in preparation for division. J Bacteriol, 1975 Mar, 121(3), 1173 - 9 Degradation of phospholipid and release of diglyceride-rich membrane vesicles during protoplast formation in certain gram-positive bacteria; Kusaka I; Membrane phospholipid was found to be hydrolyzed presumably by an intracellular phospholipase C, and diglyceride-rich membrane vesicles were released from the cells during protoplast formation in Bacillus cereus Bacillus subtilis, Micrococcus lysodeikticus, and Staphylococcus aureus . The released membranes consisted mainly of small vesicles of 50 to 100 nm in diameter . They have a lower density than that of protoplast membranes in all the bacteria tested in the present study. J Bacteriol, 1975 Mar, 121(3), 1166 - 72 Cell division suppression in the Bacillus subtilis div IC-A1 minicell-producing mutant; Mendelson NH; Growth and division patterns of Bacillus subtilis wild-type (div IV-A1+) and minicell-producing mutant (div IV-A1) clones were studied after spore germination during microcolony development in chambers that facilitate continuous observation with a phase contrast microscope . Data obtained from 13 div IV-A1+ clones were used to derive the equation DE equals {(mum minus 17.6)/8.8}, which expresses the relationship of cell divisions present in clones of various lengths . This equation was used to determine the number of divisions expected in div IV-A1 clones if the mutant clones were able to divide as often as wild-type clones . The observed number of divisions present in mutant clones was found to be only 25.27% of the number expected on the basis of this equation . Although individual div IV-A1 clones varied in the percentage of division equivalents expressed, there appeared to be no correlation between the overall clone growth rate and the number of divisions expressed . Culturing div IV-A1+ and div IV-A1 clones together in the same growth chamber revealed that there were no diffusible interactions influencing the division phenotypes of either mutant or wild-type cells . At later stages of growth, mixed microcolonies containing cells of both genotypes were formed . A length analysis of individual cells in these populations indicated that the relative division suppression of mutant compared with wild-type cells characteristic of the initial stages of clone development was maintained . It is likely, therefore, that the excessive length of minicell-producing cells (div IV-A1) is a reflection primarily of division suppression in the mutant and not simply of mislocation of division along cell length. J Bacteriol, 1975 Mar, 121(3), 917 - 22 Degradative acetolactate synthase of Bacillus subtilis: purification and properties; Holtzclaw WD et al.; A degradative acetolactate synthase (acetolactate pyruvate-lyase {carboxylating}, EC 4.1.3.18) from Bacillus subtilis has been partially purified and characterized . The synthesis of the enzyme was induced by growth of cells in minimal medium plus isobutyrate or acetate . The enzyme was partially purified by ammonium sulfate fractionation, gel filtration, and hydroxyapatite chromatography . The pH optimum of the purified enzyme was 7.0 in phosphate buffer . When assayed in phosphate buffer (pH 7.0), activity was stimulated by acetate and inhibited by sulfate . When assayed in acetate buffer (pH 5.8), activity was inhibited both by sulfate and phosphate . Michaelis-Menten kinetics was observed when the enzyme was assayed in phosphate buffer (pH 6.0 or 7.0), and inhibition by sulfate was competitive and activation by acetate was noncompetitive . When assayed in acetate buffer (pH 5.8), nonlinear Lineweaver-Burk plots were obtained; inhibition by phosphate appeared to be competitive and that by sulfate was of the mixed type . The approximate molecular weight of the purified enzyme was 250,000 as determined by gel filtration. J Bacteriol, 1975 Mar, 121(3), 807 - 13 Relation between reduced nicotinamide adenine dinucleotide oxidation and amino acid transport in membrane vesicles from Bacillus subtilis; Bisschop A et al.; The rate of reduced nicotinamide adenine dinucleotide (NADH) oxidation by membrane vesicles from Bacillus subtilis W23 increases three- to fourfold during logarithmic growth, reaching maximal levels in early stationary phase . Initial rates of L-proline, L-alanine, and L-glutamate transport energized by NADH closely parallel the increase in NADH oxidation . In vesicles prepared at different stages of growth, a constant number of NADH molecules varying from 150 to 260 have to be oxidized to transport one molecule of amino acid . Membrane vesicles from B . subtilis aroD (strain RB163), a mutant defective in menaquinone synthesis, do not transport amino acids in the presence of NADH . Ascorbate plus phenazine methosulfate, however, energizes amino acid transport equally well as in vesicles of B . subtilis W23 . NADH oxidation and NADH-driven amino acid transport can be restored instantaneously by the addition of menadione (vitamin K3). J Bacteriol, 1975 Mar, 121(3), 970 - 4 Regulation of dihydrodipicolinate synthase and aspartate kinase in Bacillus subtilis; Vold B et al.; The regulation of dihydrodipicolinate synthase (EC 4.2.1.52) and aspartate kinase (EC 2.7.2.4) was studied in Bacillus subtilis 168 . Starvation for lysine gave depression of one aspartate kinase isoenzyme but not of dihydrodipicolinate synthase . Strains resistant to growth inhibition by the lysine analogue thiosine exhibited constitutively derepressed synthesis of one aspartate kinase isoenzyme but had normal levels of dihydrodipicolinate synthase . The data provide strong evidence that lysine is not the signal for derepression of dihydrodipicolinate synthase . Nevertheless, dihydrodipicolinate synthase specific activity increased during sporulation, and it is suggested that this increase may result, in part, from resistance to proteolysis of that enzyme. J Antibiot (Tokyo), 1975 Mar, 28(3), 229 - 36 Mechanism of DNA degradation induced by neocarzinostatin in Bacillus subtilis; Ohtsuki K et al.; When logarithmically growing Bacillus subtilis cells were exposed to the antitumor protein, neocarzinostatin (NCS), at a concentration of 50 mug/ml, cellular DNA was gradually degraded into an acid-soluble form (up to 60 percent of total DNA) . The degradation appeared to initiate at the growing regions of DNA and to proceed sequentially from the nascent regions to preexistent DNA . Concomitantly with, or perhaps as a consequence of, the degradation of growing regions, DNA detached from the cell membrane and started to show single-strand nicks within 30 minutes after exposure of the cells to NCS, whereas double strand scission in the DNA became detectable in about 90 minutes . Such endonucleolytic breaks in DNA eventually gave rise to the formation of double-stranded DNA fragments of a single-size class (30-S) as determined by sedimentation in either neutral or alkaline sucrose gradients . In contrast to previous results with Sarcina lutea, the NCS-induced DNA degradation was stimulated by chloramphenicol in B . subtilis and the DNA fragment were not the final breakdown products, but were further degraded into acid-soluble materials. Clin Allergy, 1975 Mar, 5(1), 25 - 31 Observations on biphasic bronchial reactions due to inhalation of enzymes of Bacillus subtilis; Dijkman JH; The effect of disodium cromoglycate, thiazinamium and prednisone on immediate and late type allergic bronchial reactions was studied in four patients with biphasic bronchial obstruction due to inhalation of Bacillus subtilis enzymes . The immediate reaction (type 1 allergy) was not influenced by any of these drugs . Disodium cromoglycate seemed to ameliorate the late reaction, which was very much depressed by prednisone . Thiazinamium, an antihistamine drug, did not alter the immediate nor the late phase of the reaction . The data suggest that the pathogenetical mechanism involved in Bacillus subtilis allergy may differ from that initiating bronchial reactions to pollen or house dust. Eur J Biochem, 1975 Feb 21, 51(2), 587 - 91 Synthesis in vitro of phi29-specific early proteins directed by phage DNA; Carrascosa JL et al.; The RNA and proteins synthesized in an Escherichia coli cell-free system of protein synthesis directed by Bacillus subtilis phage phi29 DNA were studied . Hybridization-competition experiments showed that most of the RNA species synthesized in vitro are early RNAs . Many of the early proteins induced after phage infection were also synthesized in the E . coli cell-free system . None of the late proteins, structural or non-structural, were synthesized in the system in vitro. Eur J Biochem, 1975 Feb 21, 51(2), 503 - 10 Existence of two alternative pathways for fructose and sorbitol metabolism in Bacillus subtilis Marburg; Delobbe A et al.; Strains of Bacillus subtilis mutated for fructose phosphotransferase system (fruA), fructose-1-phosphate kinase (fruB), fructokinase (frucC) have been tested for their catabolism of sorbitol and fructose . It is shown that the previously known pathways of sorbitol and fructose degradation in B . subtilis, e.g.: (see article) may metabolize intracellular fructose produced either by sorbitol oxidation or by fructose-1-phosphate dephosphorylation . The intracellular fructore degradation via fructose-1-phosphate kinase has been found to require the fructose phosphotransferase system which ensures a vectorial transport of fructose. Eur J Biochem, 1975 Feb 21, 51(2), 415 - 27 Regulatory properties of purified 3-phosphoglycerate dehydrogenase from Bacillus subtilis; Saski R et al.; 3-Phosphoglycerate dehydrogenase (3-phosphoglycerate:NAD oxidoreductase, EC . 1.1.1.95) was purified from Bacillus subtilis by conventional methods . The final preparation was homogeneous by electrophoretic analysis and had a sedimentation constant of 6.3 S . On the basis of gel filtration data the enzyme had a molecular weight of about 166000 . The plot of velocity versus phosphoglycerate concentration was biphasic while similar plots for hydroxypyruvate phosphate and NADH were the conventional hyperbolic type . The enzyme was specifically inhibited by serine . The inhibition was time dependent, requiring several minutes incubation before a constant level of inhibition was achieved . Serine inhibition was of the "mixed type" with respect to 3-phosphoglycerate and Hill plots of these data had slopes that approached 2 . Desensitization of the enzyme to serine inhibition was achieved by incubation in the absence of dithiothreitol . The desensitized enzyme was different from the native enzyme in fluoresence properties, sedimentation characteristics and in the absence of the biphasic phosphoglycerate saturation curve . Evidence was obtained for the participation of sulphydryl groups in the changes in protein structure responsible for serine inhibition as well as the dehydrogenase activity of the enzyme. J Antibiot (Tokyo), 1975 Feb, 28(2), 126 - 8 Isolation of a new peptide antibiotic TL-119 . Studies on antibiotics from the genus Bacillus . IV; Shoji J et al.; A new antibiotic TL-119 active against gram-positive bacteria was isolated from a strain resembling Bacillus subtilis . The antibiotic is a neutral substance, soluble in a mixture of chloroform and methanol, and is a peptide with an empirical formula of C42H57N7O9, containing threonine (1), alanine (1), valine (1), leucine (1) and phenylalanine (2). J Bacteriol, 1975 Feb, 121(2), 726 - 34 Organization of teichoic acid in the cell wall of Bacillus subtilis; Birdsell DC et al.; The phytohemagglutinin, concanavalin A (Con A), interacts specifically and reversibly with the polyglucosyl glycerol phosphate teichoic acid of Bacillus subtilis 168 cell walls . Advantage has been taken of this interaction to examine the organization of the surface teichoic acid at the ultrastructural level . Con A-treated whole cells and cell walls contain an irregular, fluffy layer 25 to 60 nm thick which is absent in untreated or alpha-methyl glucoside-treated preparations . This discontinuous layer is present only on the outer profile of Con-A-treated cell walls . The surface teichoic acid is proposed to be oriented perpendicular to the long axis of the cell . Fixation and embedment for electron microscopy result in condensation of this layer which then contributes to the stainable portion of the wall . Con A treatment binds adjacent teichoic acid molecules in their native configuration producing the irregular, fluffy layer visualized. J Bacteriol, 1975 Feb, 121(2), 688 - 94 Genes affecting the productivity of alpha-amylase in Bacillus subtilis Marburg; Sekiguchi J et al.; Genetic control of alpha-amylase (alpha-1,4-glucan glucanohydrolase, EC 3.2.1.1.) production by Bacillus subtilis 168 was studied from the standpoint that alpha-amylase production by bacteria is dependent on a long-lived messenger ribonucleic acid and obeys the following equation: E = kappa integral of X-DT where x = cell mass at time t, E = alpha amylase produced, t = culture time, and kappa = productivity constant . So a productivity constand (kappa) is obtained from the slope of the straight line plot of alpha-amylase formed versus the total mass of cells accumulated over that time during the culture process . The following results were obtained . (i) Two sequential mutants, derived from the 168(kappa = 20) strain and having improved alpha-amylase productivity (168 leads to 196), were analyzed for their serine and metal protease production . Strain 128 (kappa = 40) produced half the amount of both proteases, but strain 196 (kappa = 60 similar to 80) produced 20 times that in the original strain . (ii) Amy+ transformants, using the 196 strain as the other three had higher productivity (kappa = 37 similar to 46) . These transformants (J71, J47, groups . Seventy-one of 74 Amy+ transformants had a kappa value of 21.0 plus or minus 2.1 and the other three had higher productivity (kappa = 37 similar to 46) . These transformants (J71,J47, and J10) produced levels of serine and metal proteases 20 times higher than the other transformants . (iii) Strains 196, J71, J47, and J10 were found to be nonmotile and resistant to phage PBS1, whereas other strains, including strains 168, 128, 3 revertants of strain J71 and 2 revertants of strain 196, were all motile and sensitive to the phage . (iv) Strains 196 and J71 were nonflagellated under electron microscopic observation but strain 168, 128 and a revertant of J71 were flagellated . From the above experimental results, the existence of a quality controlling gene (amyB) was deduced, which is loosely linked to the structural gene and controls productivities of alpha-amylase and proteases, and flagellation . The probable existence of another regulatory gene, amyC, is also discussed. J Bacteriol, 1975 Feb, 121(2), 450 - 4 Properties of Bacillus subtilis mutants temperature sensitive in germination; Galizzi A et al.; A new mutant of Bacillus subtilis defective in the outgrowth phase of spore germination has been isolated . When incubated at 46 C, the spores of the mutant gave rise to abnormally large swollen cells . Genetic crosses show that the mutant is different from the three previously described . The genetic analysis indicates two regions of the B . subtilis chromosome involved in the control of the spore outgrowth. J Bacteriol, 1975 Feb, 121(2), 411 - 5 Criteria for categorizing early biochemical events occurring during sporulation of Bacillus subtilis; Dancer BN et al.; Two criteria are suggested for assessing the relevance of biochemical events occurring early in sporulation . The first is thymidine starvation, a condition known to inhibit sporulation . This also inhibits the production of metalloprotease, serine protease, and ribonuclease; alpha-amylase production, however, is unaffected . The second is the effect of a regulator mutation which increases the production of the proteases . In the mutant, ribonuclease is produced in correspondingly large quantities whereas alpha-amylase production is unaffected . We conclude that, whereas the serine protease is part of the main sequence of events leading to formation of the spore, the metalloprotease is a side effect, i.e., connected with the main sequence but not part of it . Ribonuclease could, on present evidence, be either in the main sequence or a side effect associated with it . Amylase, however, seems to be separately regulated and neither directly nor indirectly connected with the sporulation sequence. J Bacteriol, 1975 Feb, 121(2), 406 - 10 Production and possible function of serine protease during sporulation of Bacillus subtilis; Dancer BN et al.; The production of extracellular protease during sporulation in Bacillus subtilis 168 was investigated . Two proteases are produced, an alkaline serine protease and a neutral metalloprotease . In vivo inhibition of the serine protease with phenylmethylsulfonylfluoride indicated that the metalloprotease was degraded by the serine protease during sporulation . The experiments with phenylmethylsulfonylfluoride also show that the serine protease is necessary for the sequential process of sporulation and that it is required continuously for the first 2 to 3 h of the 8-h process. Can J Microbiol, 1975 Feb, 21(2), 205 - 12 Structural features determining the antibiotic potencies of natural and synthetic hop bitter resins, their precursors and derivatives; Schmalreck AF et al.; Twenty-six hop bitter resins, some hitherto not investigated, were tested for antimicrobial activities . Gram-positive bacteria were much more sensitive than Gram-negative ones . The inhibitory effect against Bacillus subtilis 168 was measured by several methods and the general rule could be established that the antibiotic properties are mainly dependent on the hydrophobic parts of the molecules . Thus the acyl-lupuphenones (2-acyl-3,5-4,4',6-tri(3-methyl-2-butenyl)-cyclohexane-triones (1, 3, 5) having three prenyl and one acyl side chain are the most active substances . Their minimum inhibitory concentration (MIC) increases from the capro (0.5 muM) to the aceto derivative (11 muM) . Any substitution with hydrophilic functions or loss of hydrophobic groups causes reductions in biological activity . This is most evident with the corresponding acyl-phloroglucine precursors (2-acyl-1,3,5-trihydroxybenzenes) which lack the three prenyl side chains (MIC, 110 to 5050 muM respectively) . Conversion of the central six-membered ring structure into a five-membered one results in additional losses of antimicrobial activity . These findings support the proposal that the lipophilic region of the cell membrane represents the target site for the hop bitter resins. Biochem Genet, 1975 Feb, 13(1-2), 125 - 43 Carbamyl phosphate synthesis in Bacillus subtilis; Potvin B et al.; In vitro and "in situ" assays have been developed to test the carbamyl phosphate synthetase (CPSase) activity of a series of pyrimidine-requiring mutants of Bacillus subtilis . The enzyme has been shown to be highly unstable, and was successfully extracted only in the presence of 10% glycerol and 1 mM dithiothreitol (Cleland's reagent) . It loses activity rapidly when sonicated or when treated with lysozyme . Genetic studies, using mutants, indicate that B . subtilis may possess two CPSases . This possibility and its physiological consequences were probed enzymatically . CPSase activity has been shown to undergo inhibition by both uridine triphosphate and dihydroorotate; activation has been demonstrated in response to phosphoribosyl pyrophosphate (PRPP) and (to a lesser extent) ornithine. J Biochem (Tokyo), 1975 Feb, 77(2), 415 - 20 A new flavin enzyme catalyzing the reduction of dihydrodipicolinate in sporulating Bacillus subtilis . II . Kinetics and regulatory function; Kimura K et al.; Dihydrodipicolinate reductase in Bacillus subtilis PCI 219 had FMN as a prosthetic group, and the hydrogen transfer pathway is considered to be NADPH yields FMN yields dihydrodipicolinate . Linewaver-Burk plots of the reciprocal of the activity against the reciprocal of the concentration of either of the two substrates, dihydrodipocolinate and NADPH, are consistent with a reaction mechanism involving interconversion of two free forms of the enzyme by the two substrates . The Km values obtained from the secondary plots are 0.77 mM for dihydrodipicolinate and 72 muM for NADPH . Inhibition by dipicolinate is competitive with NADPH and noncompetitive with dihydrodipicolinate, and shows positive cooperativity . The possible metabolic role of the reductase in sporulating Bacillus subtilis is discussed in connection with regulation of the biosyntheses of dipicolinate and diaminopimelate. J Biochem (Tokyo), 1975 Feb, 77(2), 405 - 13 A new flavin enzyme catalyzing the reduction of dihydrodipicolinate in sporulating Bacillus subtilis I . Purification and properties; Kimura K; A dihydrodipicolinate reductase containing flavin was purified from sporulating Bacillus subtilis PCI 219 . The purified enzyme appeared homogeneous by dise gel electrophoresis . Its molecular weight was estimated as 74,000 by gel filtration on Sephadex G-200, and as 18,500 by electrophoresis on sodium dodecylsulfate polyacrylamid gel . These results suggest that the enzyme is composed of four subunits . The prosthetic group was identified as FMN, and one mole of the enzyme contained two moles of FMN . Both NADPH and NADH acted as coenzyme, though NADH was less effective . The enzyme also exhibited diaphorase activity . The pH optimum was 6.1 . The enzyme was inhibited by dipicolinate but not by lysine or alpha, epsilon-diaminopimelate. Mutat Res, 1975 Feb, 27(2), 157 - 69 Effects of DNA-polymerase-defective and recombination-deficient mutations on the ultraviolet sensitivity of Bacillus subtilis spores; Munakata N et al.; The DNA of UV-irradiated Bacillus subtilis spores, which contains 5-thyminyl-5,6-dihydrothymine (TDHT) as the major thymine photoproduct, is known to be repaired during germination by two complementary mechanisms: (I) the well-known excision repair, and (2) a special process, "spore repair", which destroys TDHT in situ without rendering it acid-soluble . In the absence of both mechanisms TDHT is not removed, and spores are highly UV-sensitive . When either of two mutations (pol-59 and pol-151) giving defective DNA polymerase, or one (rec-A1) giving a recombination deficiency are introduced into strains defective in one of these known TDHT removal processes, the chemically measured elimination of TDHT from spore DNA is unaltered, but spore UV-sensitivity is increased . The pol mutations produce their greatest sensitivity increase in spores of strains already deficient for the in situ destruction of TDHT, while the rec mutation gives its maximum sensitivity increase to spores of strains lacking excision . These facts argue that the pol mutations interfere mostly with excision repair (presumably its later resynthesis step), shile the rec mutation impairs "spore repair" in some step occurring subsequent to the TDHT destruction in situ . With either of these impairments of the later repair steps, DNA of UV-irradiated and germinated spores is considerably degraded, unless germination is carried out in the presence of chloramphenicol. J Virol, 1975 Feb, 15(2), 363 - 71 Inhibition of bacteriophage PBS2 replication in Bacillus subtilis by phleomycin; Post L et al.; Phleomycin is an effective inhibitor of the replication of Bacillus subtilis bacteriophage PBS2, whose DNA contains uracil instead of thymine . Phleomycin does not affect the induction of the known phage enzymes involved in deoxyribonucleotide metabolism . But phage DNA synthesis is severely inhibited by phleomycin, and late virion protein synthesis is eliminated . These effects appear to result from a phleomycin-induced degradation of the parental phage DNA . Similar inhibitory and degradative effects on DNA are seen in phleomyinc-treated, uninfected cells . This system is unaffected by the related antibiotic, bleomycin. J Biol Chem, 1975 Jan 25, 250(2), 522 - 6 Inhibition of Bacillus subtilis deoxyribonucleic acid polymerase III by phenylhydrazinopyrimidines . Demonstration of a drug-induced deoxyribonucleic acid-enzyme complex; Clements JE et al.; The interaction of 6-(phenylhydrazino)-pyrimidines and Bacillus subtilis DNA polymerase III was examined in experiments exploiting agarose gel filtration of mixtures of drug, DNA, and purified enzyme . 6-(p-Hdroxyphenylhydrazino)-uracil and 6-(p-hydroxyphenylhydrazino)-isocytosine were used as model inhibitors; both drugs induced the formation of a distinct polymerase-DNA complex . Comples formation required the inhibitory, hydrazino forms of the drugs and a form of DNA suitable as a primer-template for DNA polymerase III . dGTP and dATP, which respectively, competitively antagonize the inhibitory effects of the uracil and isocytosine derivatives, antagonized in an equally specific manner the respective capacities of these compounds to induce complex formation . Experiments exploiting both wild type and drug-resistant, mutant polymerases indicated that drug concentrations required for the half-maximal induction of complex formation were nearly identical with the apparent inhibitor constants (Ki) determined independently by kinetic analysis of enzyme inhibition . These results and those of experiments exploiting defined homopolymer-oligomer combinations as template-primers support a model of inhibitor action in which arylhydrazinopyrimidine forms a reversible, ternary complex with the enzyme and an appropriate timplate pyrimidine residue in an area adjacent to the 3-hydroxyl primer terminus. Biochim Biophys Acta, 1975 Jan 20, 378(2), 171 - 85 Retardation time measurementson replicating bacillus subtilis chromosomes: effect of EDTA concentration; Muller WA et al.; We have found that high concentrations of EDTA (greater than 0.024 M) are necessary to produce large, constant numbers of intact replicating Bacillus subtilis chromosomes in lysates of log phase cells . The retardation time of replicating chromosomes in log phase cell lysates is about double that for chromosomes in stationary phase cell lysates, thus making measurement of retardation time a sensitive way to detect and study replicating chromosomes . A theory is developed to predict retardation times for many possible models of DNA replication . The retardation time data on log phase cells is sufficient to eliminate many replication models, but many possibilities remain. Biochim Biophys Acta, 1975 Jan 6, 378(1), 35 - 43 Deoxyribonucleic acid synthesis induced with ultraviolet light in Brij 58-treated Bacillus subtilis spores germinated in the presence of chloramphenicol; Fujita Y et al.; The direct measurement of ultraviolet light-stimulated DNA synthesis in the permeable Bacillus subtilis cells was performed . Bacillus subtilis spores germinated in the presence of chloramphenicol were treated with Brij 58 and irradiated with ultraviolet light, and (3H)dTTP was incorporated into these cells by the DNA polymerase assay system . Characteristics of the incorporation were distinct from those into spores germinated in the absence of chloramphenicol and treated with Brij 58, in the respect that the former incorporation did not require ATP and only partially depended on the presence of all four deoxyribonucleoside triphosphates . The incorporation of (3H)dTTP into DNA was confirmed by CsCl density gradient centrifugation . A DNA polymerase I-deficient strain, JBl 49(59) had no (3H)sTTP incorporating activity induced by ultraviolet light irradiation when the germinated spores were treated with Brij 58 . Analysis of alkaline sucrose gradient centrifugation revealed that fragmented DNA caused by ultraviolet light irradiation was rejoined to the size of DNA of non-irradiated cells by incubating irradiated cells in the DNA polymerase assay mixture containing NAD+ . The results also suggested that a machinery of DNA repair probably pre-existed in the spore. Genetika, 1975, 11(8), 77 - 80 {UV-mutagenesis in Bacillus subtilis . IV . Analysis of revertants to methionine prototrophy}; Fillippov VD et al.; UV light induces in Bacillus subtilis met5 ade6 two classes of revertants to prototrophy to methionine which can be easily distinguished by their phenotype: double (Met+Ade+) and solitary (Met+) revertants . Crosses of revertants with the wild type, carried out in transformational experiments, showed that original (direct) mutation met5 is presented in chromosome of double revertants . Consequently they are extragenic suppressor revertants . In the chromosome of solitary revertants Met+ an extragenic suppressor was not detected; reversions Met+ seem to be of an intragenic nature . It is possible to use reversions to prototrophy to methionine as a model to study UV-mutagenesis in suppressor and non-suppressor genes. Genetika, 1975, 11(12), 146 - 9 {Clonal analysis of the progeny of UV-irradiated Bacillus subtilis (uvr+ and uvr-) cells}; Lotareva OV et al.; The revertants to adenine prototrophy or mutants to auxotrophy can be easily identified on synthetic media which are partly enriched by caseine hydrolysate and yeast extract . It is shown with the use of these media that 1.5% colonies formed by Bacillus subtilis cells of the original type (ade6 met5) have mutant clones which are initiated by spontaneous revertants to adenine prototrophy . These revertants arise in the time of division of cells in macrocolonies . After plating diluted suspension of irradiated cells those colonies which contain mutant clones formed by spontaneous revertants can be erroneously taken for mixed colonies formed by induced revertants . About 40% mutants to auxotrophy induced by high dose of UV-light in in uvr+ cells form pure mutant colonies . The same mutants, induced by uvr cells by five time less UV-dose, usually form mixed colonies. Antonie Van Leeuwenhoek, 1975, 41(4), 513 - 9 Mating reaction in Saccharomyces cerevisiae . VII . Effect of proteolytic enzymes on sexual agglutinability and isolation of crude sex-specific substances responsible for sexual agglutination; Shimoda C et al.; The effect of proteolytic enzymes on sexual agglutinability of haploid cells of the yeast Saccharomyces cerevisiae was examined . Sexual agglutinability of cells of both a and alpha types was lost on treatment with alkaline protease and two kinds of neutral proteases of Bacillus subtilis, pronase and alpha-chymotrypsin . Agglutinability of alpha type cells was lost after treatment with acid protease of Rhizopus chinensis and trypsin, but that of a type cells was not . These results indicate that the sex-specific substance responsible for the sexual agglutination (agglutination factor) in a type cells differ from that in alpha type cells . Agglutination factors were solubilized from cell-wall fractions of both mating types by Glusalase treatment . These crude factors specifically inhibited the agglutinability of cells of the opposite mating type with little effect on the agglutinability of cells of the same mating type. Genetika, 1975, 11(8), 129 - 38 {Operon of riboflavin synthesis in Bacillus subtilis . IX . Preparation and properties of lumiflavin- or lumichrome-resistant mutants}; Bresler SE et al.; Lumiflavin and lumichrome both inhibit the growth of different strains of Bacillus subtilis including riboflavin-constitutive mutant . Lumiflavin has no regulating effect, but lumichrome is an effector and it participates in the repression of riboflavin precursors synthesis and the synthesis