d-glucose dehydrogenase from bacillus megaterium m 1286: purification, properties and structure.1) glucose dehydrogenase from bacillus megaterium has been purified to a specific activity of 550 u per mg protein. the homogeneity of the purified enzyme was demonstrated by gel electrophoresis and isoelectric focusing. 2) the amino acid composition has been determined. 3) the molecular weight of the native enzyme was found to be 116000 by gel permeation chromatography, in good agreement with the values of 120000 and 118000, which were ascertained electrophoretically according to the method of ...19752530
involvement of a single hydroxylase species in the hydroxylation of palmitate at the omega-1, omega-2 and omega-3 positions by a preparation from bacillus megaterium.a soluble enzyme preparation from bacillus megaterium, requiring nadph and o2 for activity and containing ferredoxin-replaceable and cytochrome p-450-type components, was previously shown to catalyze the conversion of palmitic acid to an isomeric mixture of omega-1, omega-2 and omega-3 hydroxypalmitate. it has now been shown that the ratio of these three positional isomers in the enzymatic product remains unchanged in spite of partial diminution of total hydroxylase activity by heat treatment, p ...19767299
physiological state of bacillus megaterium in the chemostat under alkaline conditions. 19769737
growth of bacillus megaterium in the chemostat under alkaline conditions. 19769738
effects of added germination agents on loss of optical density in electron-irradiated spores.spores of bacillus megaterium atcc 14581, subjected to partial-cell iradiation, were exposed to either lysozyme, h2o2, or glucose in an attempt to reduce or eliminate the nonmonotonic behavior in curves of percentage of germination versus energy, obtained when such spores were resuspended in phosphate buffer alone. except at the lower doses. h2o2 effectively eliminated this anomalous dip in these curves, whereas lysozyme amplified it greatly. glucose was generally ineffective. coinciding with th ...19769902
membrane-bound dd-carboxypeptidase and transpeptidase activities from bacillus megaterium km at ph 7. general properties, substrate specificity and inhibition by beta-lactam antibiotics.1. the membranes from bacillus megaterium km contained a dd-carboxypeptidase with optimum activity under the following conditions: ph 7; ionic strength, 1.3 m; temperature, 40 degrees c and below 20 degrees c. it did not require any divalent cation, but was inactivated by cu2+ and hg2+. it was stimulated by 2-mercaptoethanol and low concentrations of p-chloromercuribenzoate. 2. the membrane preparation also catalyzed a simple transpeptidation reaction using as carboxyl acceptors d-alanine or gly ...197610161
levels of oxidized and reduced pyridine nucleotides in dormant spores and during growth, sporulation, and spore germination of bacillus megaterium.dormant spores of bacillus megaterium contained no detectable reduced nicotinamide adenine dinucleotide (nadh) or reduced nicotinamide adenine dinucleotide phosphate (nadph) despite significant levels of the oxidized forms of these nucleotides (nad and nadp). during the first minutes of spore germination there was rapid accumulation of nadh and nadph. however, this accumulation followed the fall in optical density that is characteristic of the initiation of spore germination. accumulation of nad ...197714113
studies on the mechanism of interaction of glutaraldehyde with microorganisms (author's transl).the antimicrobial action of glutaraldehyde on candida lipolytica increases with reagent concentration, ph and duration of contact. the simultaneous study of the size distribution of the particles shows an agglutination of the cells. this result is confirmed by direct observation of the cells by electron microscopy. cell agglutination also occurs with other microorganisms (saccharomyces carlsbergensis, bacillus megaterium, escherichia coli) and increases their settling rate. the formation of such ...197614579
physiologo-biochemical properties of a chemostatic culture of bacillus megaterium at different ph of the chemostat culture of bacillus megaterium were studied during its growth in the state of stress caused by unfavourable acid and alkaline values of ph. the effect of ph is not specific since changes occur in both energy exchange and constructive metabolism. the common action of hydrogen and hydroxyl ions is a decrease in the economic coefficient as greater amounts of the energy substrate are being used, the repression of oxidative enzymes, a decrease in the pool of atp, and an in ...197716198
activities of anaplerotic enzymes and acetyl coenzyme a carboxylase in biotin-deficient bacillus megaterium. 197717649
characteristics of a cytochrome p-450-dependent fatty acid omega-2 hydroxylase from bacillus megaterium.the fatty acid (omega-2) hydroxylase from bacillus megaterium atcc 14581 was examined with respect to some general enzymatic properties attributed to an intact complex isolated in a partially purified state. hydroxylase specific activity was found to increase with increasing protein concentration in a manner consistent with a reversible association of the components in the complex. there was a substantial kinetic lag phase for palmitate hydroxylation which was abolished by a substrate preincubat ...197718202
dihydrodipicolinate reductases from bacillus cereus and bacillus megaterium.dyhydrodipicolinate reductases were purified 100-fold from crude extracts of b. cereus and b. megaterium and their properties were compared with those of the reductase from b. subtilis. the molecular weights of the reductases of b. cereus and b. megaterium were fount to be 155,000 and 150,000, respectively. these reductases were shown to be free of flavin, unlike the b. subtilis enzyme, which contains flavin. both nadph and nadh acted as coenzymes for these two reductases. nadph being three or f ...197719431
chemical composition of cells of a chemostatic culture of bacillus megaterium during exposure to an alkaline ph value.the culture of bacillus megaterium was grown under chemostat conditions at a rate of 0.2, 0.4 and 0.7 hr-1, the growth being limited by a low content of citrate, and at alkaline ph values. critical ph values were obtained for each growth rate: 9.6, 9.2 and 7.6, respectively. the content of protein decreased at alkaline ph, while the synthesis of total lipids, poly-beta-hydroxybutric acid and phospholipids was stimulated. some changes were found in the ratio between polyphosphates of high molecul ...197719686
a unique method for studying the initiation of bacillus megaterium spore germination. 197723775
enolase from spores and cells of bacillus megaterium: two-step purification of the enzyme and some of its properties.a simple two-step procedure for purification of enolase from germinated spores or vegetative cells of bacillus megaterium is described. the procedure resulted in a 1,200-fold purification with production of homogeneous enzyme in approximately 75% yield; the enzymes from spores and cells seemed identical. the molecular weight of the native enzyme was 335,000, with a subunit molecular weight of 42,000. the enzyme required mg2+ and was inhibited by ethylenediaminetetraacetic acid and fluoride ions. ...197825885
metal content in bacillus megaterium cells under different cultivation conditions.the content of k, na, mg, fe, al, ba, sr and cu was studied in the cells of a chemostal culture of bacillus megaterium whose growth was limited with citrate, the growth rates being d=mu=0.2, 0.4 and 0.7 hr(-1). the growth was affected also by acid and alkaline values of ph. the content of metals in the cells could change tenfold, depending on their physiological state. the content of ca, mg and fe increased with the growth rate while that of k remained constant. the content of metals changed eve ...197826013
an evaluation of respiration chain-associated functions during initiation of germination of bacillus megaterium bacillus megaterium qm b1551, spore germination could be initiated by glucose in the absence of detectable oxygen consumption, atp synthesis or a ph decrease in the external media, suggesting that none of those reactions were mandatory. in addition, initiation of germination was insensitive to a variety of inhibitors of energy production or protonmotive force uncouplers. therefore the respiratory chain-associated functions are not prerequisites for initiation of germination but these function ...197827232
relationship between sporulation and mutations impairing glutamine synthetase in bacillus megaterium. 197528123
inactivation of glutamate dehydrogenase and glutamate synthase from bacillus megaterium by phenylglyoxal, butane-2,3-dione and pyridoxal 5'-phosphate.reaction of phenylglyoxal with glutamate dehydrogenase (ec, but not with glutamate synthase (ec, from bacillus megaterium resulted in complete loss of enzyme activity. nadph alone or together with 2-oxoglutarate provided substantial protection from inactivation by phenylglyoxal. some 2mol of 14cphenylglyoxal was incorporated/mol of subunit of glutamate dehydrogenase. addition of 1mm-nadph decreased incorporation by 0.7mol. the ki for phenylglyoxal was 6.7mm and ks for competit ...197828736
membrane bioenergetic parameters in uncoupler-resistant mutants of bacillus megaterium.mutants of bacillus megaterium displaying malate-driven atp synthesis resistant to uncouplers of oxidative posphorylation are further characterized. both the ph gradient and electrical potential generated across the membrane by malate respiration are equally sensitive to uncouplers in the wild type and uncoupler-resistant mutants. the mutants possess 0 to 10% of the wild type atpase activity which is not activated by pretreatment with heat or trypsin. despite this inability to measure atpase act ...197829041
effect of the redox potential on the growth of aerobic microorganisms.the effect of redox potential was studied on the growth of the following aerobic microorganisms: candida utilis, bacillus megaterium, pseudomonas fluorescens. the action of oxidizing agents (k3fe(cn)6, kio3, k2cr207 and kmno4) and reducing agents (ascorbic acid, sodium thioglycolate, k4fe (cn)6 and na2s2o3) on the growth rate was investigated. k3fe(cn)6, ascorbic acid, sodium thioglycolate and k4fe(cn)6 were found to be suitable for buffering the eh of the medium. the potential could be shifted ...197830023
purification and properties of phosphoglycerate phosphomutase from spores and cells of bacillus megaterium.phosphoglycerate phosphomutase has been purified to homogeneity from vegetative cells and germinated spores of bacillus megaterium, and the spore and cell enzymes appear identical. the enzyme is a monomer of molecular weight 61,000. the compound 2,3-diphosphoglyceric acid is not required for activity, but the enzyme has an absolute and specific requirement for mn2+. the enzyme is inhibited by ethylenediaminetetraacetate and sulfhydryl reagents, has a ph optimum of about 8.0, and has km values fo ...197933959
pterin deaminase from bacillus megaterium. purification and properties.a pterin deaminase catalyzing the hydrolytic deamination of various pteridines was found in the bacterium, bacillus megaterium, and partially purified from bacterial extract. the specific activity was raised 90-fold over that of the crude extract. the ph optimum is around 7.3, and the km value for 6-carboxypterin is 1.3 mm. the molecular weight of the enzyme was estimated by gel filtration to be about 110,000. the enzyme deaminated pterin, 6-carboxypterin, biopterin, 6-methylpterin, 7-methylpter ...197934599
biochemistry of l-proline-triggered germination of bacillus megaterium spores.the mechanism by which l-proline triggers germination in bacillus megaterium qm b1551 spores was investigated. first, brief exposure of spores to l-proline, followed by dilution, was sufficient to trigger germination. once germination was triggered, the spores continued initiation of germination and did not require high concentrations of l-proline. triggering of germination was ph and temperature dependent. second, enzymes for l-proline catabolism were absent in spores, and several non-metaboliz ...197935526
effect of dicumarol on the growth of some soil microorganisms.the effect of dicumarol on growth of selected soil bacteria: azotobacter chroococcum, arthrobacter globiformis, a. citreus and bacillus megaterium was studied. the following minimum concentrations were inhibitory in vitro: arthrobacter citreus--20 mug/ml., bacillus megaterium--40 mug/ml., azotobacter chroococcum--40 mug/ml. arthrobacter globiformis--70 mug/ml. cells of all microorganisms studied grown in the presence of dicumarol developed aberrant morphological forms.197659533
immunochemical study of the peptidoglycan of gram-negative bacteria.the specificity of antibodies directed against the peptidoglycan of gram-negative bacteria was studied. the peptidoglycans of proteus vulgaris, escherichia coli, moraxella glucidolytica, neisseria perflava, give identical precipitin reactions. by means of inhibition studies with various peptidoglycan subunits and synthetic peptides, it was shown that the antibodies are essentially directed against the peptide moiety of the peptidoglycan: l-ala-d-glu (l)-mesoa2pm-(l)-d-ala, that the peptide react ...197660233
purification and characterization of the penicillin-binding protein that is the lethal target of penicillin in bacillus megaterium and bacillus licheniformis. protein exchange and complex stability.the penicillin-binding protein that is thought to be the lethal target of penicillin in bacillus megaterium (protein 1) has been purified to greater than 95% homogeneity. the membrane-bound penicillin-binding proteins were solubilized with a non-ionic detergent and partially separated from each other by ion-exchange chromatography on deae-sepharose cl-6b. protein 1 was subsequently purified by covalent affinity chromatography on ampicillin-affinose. bacillus licheniformis contains an equivalent ...197897081
study of calcium dipicolinate release during bacterial spore germination by using a new, sensitive assay for dipicolinate.the release of calcium and dipicolinic acid from spores of bacillus megaterium km during l-alanine-induced triggering of germination has been studied using a new, simple, and rapid assay for dipicolinic acid capable of detecting a concentration of 0.5 micron. the release of both calcium and dipicolinate started within seconds of exposure of the spores to l-alanine, thus preceding other measurable changes associated with germination. from the earliest times, the two substances were released in eq ...197897264
polyethylene glycol-induced fusion of heat-inactivated and living protoplasts of bacillus megaterium.protoplasts of bacillus megaterium, incubated at 50 degrees c for 120 min, lost the ability to revert to bacillary form. such heat-inactivated protoplasts, however, produced recombinants when fused by polyethylene glycol treatment with normal protoplasts. although this differential inactivation effect is not yet fully reproducible, reciprocal inactivations of the parental protoplasts in genetic crosses have clearly shown that for protoplast fusion (i) either of the parents may serve as the viabl ...197897276
divalent cation mobility throughout exponential growth and sporulation of bacillus megaterium.each of the five elements considered was taken up by bacillus megaterium during exponential growth. initial mg and mn uptake was rapid and ended by mid-log. for ca, fe, and zn, uptake continued throughout exponential growth. elements were released from the cells immediately following initial uptake. for mn, egression continued to t2, with release of 36% of total accumulated. secondary uptake followed immediately and continued through stage v. magnesium egression continued to t1 with release of 3 ...197897497
effects of some hypoxic cell radiosensitizers on the decay of potentially lethal oxygen-dependent damage in fully hydrated spores.using a stopped-flow mixing and pulsed irradiation apparatus, a study has been made of the decay, to a harmless form, of radiation-induced species that would otherwise be lethal to spores on contact with oxygen. aqueous suspensions of bacillus megaterium spores were irradiated with electrons for approximately 1 s; at various times after irradiation oxygen in solution was added. as the interval between anoxic irradiation and introduction of oxygen increased, the fraction of spores surviving incre ...197898174
purification and properties of glutamate synthase and glutamate dehydrogenase from bacillus megaterium.bacillus megaterium n.c.t.c. no. 10342 exhibits glutamate synthetase (ec and glutamate dehydrogenase (ec activities. concentrations of glutamate synthase were high when the bacteria were grown on 3mm-nh4cl and low when they were grown on 100mm-nh4cl, whereas glutamate dehydrogenase concentrations were higher when the bacteria were grown on 100mm-nh4cl than on 3mm-nh4cl. glutamate synthase and glutamate dehydrogenase were purified to homogeneity from b. megaterium grown in 10mm ...197899144
lipid and protein composition of membranes of bacillus megaterium variants in the temperature range 5 to 70 degrees c.membranes were prepared from four temperature range variants of bacillus megaterium: one obligate thermophile, one facultative thermophile, one mesophile, and one facultative psychrophile, covering the temperature interval between 5 and 70 degrees c. the following changes in membrane composition were apparent with increasing growth temperatures: (i) the relative amount of iso fatty acids increased and that of anteiso acids decreased, the ratio of iso acids to anteiso acids being 0.34 at 5 degree ...197899426
isolation and characterization of bacillus megaterium mutants containing decreased levels of spore protease.a proteolytic activity present in spores of bacillus megaterium has previously been implicated in the initiation of hydrolysis of the a, b, and c proteins which are degraded during spore germination. four mutants of b. megaterium containing 20 to 30% of the normal level of spore proteolytic activity have been isolated. partial purification of the protease from wild-type spores by a reviewed procedure resulted in the resolution of spore protease activity on the a, b, and c proteins into two peaks ...197899436
relationships between intracellular proteolytic activity and protein turnover in bacillus megaterium.when incubated in a sporulation medium, the sporogenous strains of bacillus megaterium degrade proteins at a rate of 4-10% x h-1. the maximal rate of protein turnover is reached after 3-4 hrs at the time of development of forespores and then decreases again. the rate of protein turnover in the asporogenous strain decreases steadily under similar conditions from 3-8% x h-1 at the beginning of incubation to 1% x h-1 after 5-6 hrs in the sporulation medium. the rate of degradation of proteins in vi ...197799935
functional half-life of the exocellular protease mrna of bacillus megaterium.functional half-life of the exocellular protease mrna was determined in in exponentially growing and stationary cells of the asporogenic strain of bacillus megaterium km and in the sporogenic strain of b. megaterium 27 during sporulation. no reserve of the protease mrna could be detected in the cells and the half-lives were determined to be 6--7 min in the exponential and stationary cells of b. megaterium km and 7.5--8.5 min in b. megaterium 27. the mean half-life of mrna for cell proteins was d ...1978100398
[growth characteristics of a lysogenic culture of bacillus megaterium under periodic and continuous cultivation]. 1978100668
microbial catabolism of vanillate: decarboxylation to guaiacol.a novel catabolic transformation of vanillic acid (4-hydroxy-3-methoxybenzoic acid) by microorganisms is reported. several strains of bacillus megaterium and a strain of streptomyces are shown to convert vanillate to guaiacol (o-methoxyphenol) and co2 by nonoxidative decarboxylation. use of a modified most-probable-number procedure shows that numerous soils contain countable numbers (10(1) to 10(2) organisms per g of dry soil) of aerobic sporeformers able to convert vanillate to guaiacol. conver ...1978101140
metabolism and the triggering of germination of bacillus megaterium. concentrations of amino acids, organic acids, adenine nucleotides and nicotinamide nucleotides during germination.a considerable amount of evidence suggests that metabolism of germinants or metabolism stimulated by them is involved in triggering bacterial-spore germination. on the assumption that such a metabolic trigger might lead to relatively small biochemical changes in the first few minutes of germination, sensitive analytical techniques were used to detect any changes in spore components during the l-alanine-triggered germination of bacillus megaterium km spores. these experiments showed that no chang ...1978101212
metabolism and the triggering of germination of bacillus megaterium. use of l-[3h]alanine and tritiated water to detect metabolism.l-[2,3-3h]alanine was used to probe for metabolism of alanine during triggering of germination of spores of bacillus megaterium km. no detectable incorporation of label into any compound, including water, was found, indicating that any metabolism involving the alanine germinant must be at a very low rate and also that alanine racemase is absent from spores of this strain. spores were germinated in 3h2o to find if any of the many metabolic reactions causing irreversible incorporation of 3h into r ...1978101213
molecular structure of the teichuronic acid of bacillus megaterium. 1978101233
adjuvant effect of a peptidoglycan attached covalently to a synthetic antigen provoking anti-phage antibodies.the synthetic antigen denoted p2-a--l, comprising the fragment p2 of the coat protein of ms-2 coliphage attached to multichain poly-dl-alanine, served for the immunization of guinea-pigs. immunization was carried out either in phosphate buffered saline (pbs) or in freund's incomplete adjuvant (fia) in the presence or absence of a small molecular weight peptidoglycan prepared from bacillus megaterium, which was checked for its adjuvant effect. the various antisera were assessed by their capacity ...1978101440
purification and characterization of additional low-molecular-weight basic proteins degraded during germination of bacillus megaterium spores.dormant spores bacillus megaterium contained a group of low-molecular-weight (5,000 to 11,000) basic (pi greater than 9.4) proteins (termed d, e, f, and g proteins) which could be extracted from disrupted spores with strong acids. these proteins were distinct from the previously described a, b, and c proteins which are degraded during spore germination. however, the d, e, f, and g proteins were also rapidly degraded during spore germination, accounting for 10 to 15% of the protein degraded. prot ...1978101515
[the effect of hgcl2 on the germination of bacillus megaterium qm b1551 spores (author's transl)]. 1978101692
effect of beta-lactam antibiotics on in vitro peptidoglycan cross-linking by a particulate fraction from escherichia coli k-12 and bacillus megaterium km.the binding constants of several beta-lactam antibiotics towards penicillin-binding components in escherichia coli k-12 (spratt, eur. j. biochem. 72:341-352, 1977) and the antibiotic concentrations required to inhibit the peptidoglycan transpeptidase of e. coli 50% were compared. penicillin-binding component 1b may have been the transpeptidase working in vitro. the structure-activity relationships of beta-lactam antibiotics and the mechanisms of action in e. coli and bacillus megaterium are disc ...1978102248
calcium accumulation during sporulation of bacillus megaterium km.accumulation of ca2+ in bacilli occurs during stages iv to vi of sporulation. ca2+ uptake into the sporangium was investigated in bacillus megaterium km in protoplasts prepared in stage iii of sporulation and cultured to continue sporulation. these protoplasts and whole cells exhibit essentially identical ca2+ uptake, which is compared with that of forespores isolated in stage v of sporulation. ca2+, uptake into both sporangial protoplasts and isolated forespores occurs by ca2+-specific carrier- ...1978103543
chromosome age and segregation during sporulation of bacillus megaterium.the effect of chromosome age on segregation during sporulation was investigated. vegetative cells of bacillus megaterium were labeled with [me-3h]thymine and then were grown at 30 degrees c in nonradioactive medium for various times before being allowed to sporulate. the ratio of the amount of label in sporal dna to that in sporangial dna, obtained after minor correction for the sporulation frequency, remained essentially constant as the postlabeling growth period was increased from one to seven ...1978103607
induced chain formation in bacillus megaterium by suramin (bayer 205).b. megaterium nctc 5637 has been shown to grow into long chains in 1% glucose and 1% suramin (w/v). this effect was not noticeable in lower concentration of the inhibitor. the effect diminished in suramin free medium, indicating the change to be phenotypic.1978104476
effects of alpha- and beta-d-glucose on germination of spores of bacillus megaterium qm b1551.the effects of d-glucose anomers on the germination of dormant spores of bacillus megaterium qm b1551 were studied, alpha-d-glucose (1 mm) slightly initiated the germination of the dormant spores during 10 min incubation at 37 degrees c, while about 60% of the dormant spores became germinated with beta-d-glucose (1 mm) in the same conditions. from the above observations and the finding that only a trace amount of alpha- or beta-d-glucose may bind with the dormant spores, it is speculated that th ...1979104990
formation of 9,10-epoxypalmitate and 9,10-dihydroxypalmitate from palmitoleic acid by a soluble system from bacillus megaterium. 1978105735
peptidoglycan turnover during growth of a bacillus megaterium dap- lys- mutant.the aim of this study was to ascertain whether or not the absence of cell wall growth zones, deduced from the analysis of autoradiographs of dl-[3h]mesodiaminopimelic acid pulse-labeled cells of a dap- lys- mutant of bacillus megaterium, was due to a high peptidoglycan turnover. turnover was determined in very precise experimental conditions because two kinds of turnover occurred: a low, acid-soluble turnover and a high, acid-insoluble one. the latter was detected during a chase in the culture m ...1979106045
[xerophytic microorganisms multiplying under conditions close to martian ones].the xerophytic cultures bacillus megaterium and mycococcus ruber can grow in the imitated martian conditions. therefore, they are resistant to the extreme factors of the environment: the composition of atmosphere, low pressure, the maximum hygroscopicity of a substrate, periodic freezing-thawing.1979106224
the primary structure of the teichuronic acid of bacillus megaterium. 1979107162
accumulation of iron by yersiniae.escherichia coli, bacillus megaterium, and three species of yersiniae grew rapidly without significant production of soluble siderophores in a defined iron-sufficient medium (20 microm fe3+). in iron-deficient medium (0.1 to 0.3 microm fe3+) all organisms showed reduced growth, and there was extensive production of siderophores by e. coli and b. megaterium. release of soluble siderophores by yersinia pestis, y. pseudotuberculosis, or y. enterocolitica in this medium was not detected. citrate (1 ...1979108243
glucose-triggered germination of bacillus megaterium spores.triggering of germination in bacillus megaterium qm b1551 spores with d-glucose was studied. first, the interaction of glucose with spores for less than 1 min resulted in triggering almost 90% of the spores after the glucose was removed by dilution. therefore only a brief time is needed for glucose to trigger germination, and then the continuous presence of glucose is not necessary. detectable uptake of glucose began 2 to 3 min after absorbance loss started, and a non-metabolizable glucose analo ...1979108260
dissociation and reconstitution of membranes synthesizing the peptidoglycan of bacillus megaterium. a protein factor for the polymerization step.cholate-solubilized bacillus megaterium membranes can be reconstituted by dialysis in the presence of magnesium ion to regain approximately 12% of the original peptidoglycan synthetic activity. bio-gel a-5m filtration of the solubilized components shows that all of the compounds necessary for peptidoglycan synthesis can be dissociated into material with a molecular weight of less than approximately 68,000. using this reconstitution system, an assay has been developed for a new protein factor, pg ...1979108266
localization of phospholipids in the membrane of bacillus megaterium. 1979109297
the direction of transpeptidation during cell wall peptidoglycan biosynthesis in bacillus megaterium. 1979109317
purification and characterization of a carboxypeptidase-transpeptidase of bacillus megaterium acting on the tetrapeptide moiety of the peptidoglycan.the enzyme carboxypeptidase-iiw of bacillus megaterium incorporates free diaminopimelate into purified bacterial walls. this enzyme can be solubilized from toluene-treated cells by licl extraction and has now been purified 106-fold to one major band on polyacrylamide gel electrophoresis. the enzyme has an apparent molecular weight of approximately 60,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by sephadex g-100 gel filtration. carboxypeptidase-iiw requires divalent catio ...1979109439
ultrastructure of the capsule of bacillus megaterium atcc 19213.the ultrastructure of the firmly adherent capsule produced by bacillus megaterium cultured on fructose mineral salts medium was examined using thin sectioning, freeze-etching, and critical point drying by transmission and scanning electron microscopy. the capsule material was shown to be fibrillar, with most fibrils containing bulbous protrusions. two types of fibres were resolved. these were termed primary and cross-linking fibres. primary fibres originated at the cell wall and had a diameter o ...1978109742
conditions for induction of bacteriophage from lysogenic bacillus megaterium with aflatoxin b1.the present study was conducted to determine whether or not aflatoxin b1 was an effective inducing agent for lysogenic bacteria and to characterize some of the parameters involved in induction. a lysogenic strain of bacillus megaterium (nrrl-b-3695) and an indicator strain of this species (nrrl-b-3694) were used. cultures of the lysogenic strain were incubated for various periods of time in the presence of aflatoxin b1. plaque-forming units as well as colony-forming units were then determined. r ...1979110264
energy-dependence of calcium accumulation during sporulation of bacillus megaterium km.ca2+ accumulation and endogenous respiration of sporulating bacillus megaterium are inhibited to the same extent by electron-transport of inhibitors and the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone, suggesting that ca2+ is accumulated by an active transport process. forespores isolated in stage v of sporulation demonstrated ca2+-specific carrier-mediated ca2+ uptake, consistent with downhill transfer [hogarth & ellar (1978) biochem. j. 176, 197-203]. in the present studies fo ...1979110319
characterization of an inhibitor of the intracellular protease from bacillus subtilis.a specific inhibitor of intracellular serylprotease from bacillus subtilis has been isolated from both growing and sporulating cells. like other protease inhibitors isolated from eukaryotic cells, the inhibitor from b. subtilis is a thermostable protein. a purification method is described. the molecular weight estimated by biogel filtration and sds gel electrophoresis is about 15,500. both proteolytic and esterolytic activities of intracellular protease are equally sensitive to inhibition. with ...1979110359
localization of low-molecular-weight basic proteins in bacillus megaterium spores by cross-linking with ultraviolet light.two low-molecular-weight basic proteins, termed a and b proteins, comprise about 15% of the protein of dormant spores of bacillus megaterium. irradiation of intact dormant spores with ultraviolet light results in covalent cross-linking of the a and b proteins to other spore macromolecules. the cross-linked a and b proteins are precipitated by ethanol and can be solubilized by treatment with deoxyribonuclease (75%) or ribonuclease (25%). irradiation of complexes formed in vitro between deoxyribon ...1979110790
identification of an nadh-linked disulfide reductase from bacillus megaterium specific for disulfides containing pantethine 4',4''-diphosphate moieties.bacillus megaterium contains an nadh-linked disulfide reductase that is specific for disulfides containing pantethine 4',4''-diphosphate moieties. this reductase is at its highest level in cells late in sporulation and in dormant spores, and could be involved in the formation and cleavage of coenzyme a-protein disulfides which take place late in sporulation and early in spore germination, respectively.1979110800
[cleavage of lactulose by bacillus megaterium]. 1978111429
transduction in bacillus megaterium. 1979111676
methylation of basic proteins in ribosomes from wild-type and thiostrepton-resistant strains of bacillus megaterium and their electrophoretic analysis.ribosomes, radioactively labelled in vivo with both [1-14c]methionine and [methyl-3h]methionine, have been isolated from both wild-type and thiostrepton-resistant strains of bacillus megaterium and their constituent proteins separated by two-dimensional gel electrophoresis. ribosomes from the wild-type strain possess one basic protein that is extensively methylated. in contrast no such protein can be detected in ribosomes from the thiostrepton-resistant strain.1979111927
binding of thiostrepton to a complex of 23-s rrna with ribosomal protein l11.thiostrepton binds with high affinity and with a 1 : 1 stoichiometry to a complex formed between escherichia coli 23-s ribosomal rna and ribosomal protein l11 of e. coli or the homologous protein bm-l11 of bacillus megaterium. in the presence of t1 ribonuclease, protein bm-l11 and thiostrepton protect from degradation a fragment of e. coli 23-s rna estimated to be about 50 nucleotides in length.1979111931
physical mapping of a plasmid from bacillus megaterium by restriction endonuclease cleavage. 1979112602
l-proline site for triggering bacillus megaterium spore germination. 1979114177
effects of apramycin, a novel aminoglycoside antibiotic on bacterial protein synthesis.1. the novel aminoglycoside antibiotic apramycin is shown to be a potent inhibitor of protein synthesis in bacteria both in vivo and in vitro. 2. in cell-free systems from escherichia coli programmed with poly(u), apramycin induces translation errors, as assayed by incorporation of leucine, isoleucine and serine, although this effect occurs only to a limited extent. 3. apramycin inhibits the translocation step of protein synthesis both in vivo, in protoplasts of bacillus megaterium, and in vitro ...1979115690
covalent structure of protein a. a low molecular weight protein degraded during germination of bacillus megaterium spores.the complete covalent structure of protein a, a protein degraded during bacterial spore germination, has been determined. the intact protein was cleaved with a highly specific spore protease into two peptides, residues 1 to 21 and 22 to 61. the larger peptide was further cleaved into two fragments with either cyanogen bromide or by trypsin cleavage following arginine modification with cyclohexanedione. the peptides derived from cyanogen bromide fragmentation encompassed residues 22 to 53 and 54 ...1979115874
requirement for ribosomal protein bm-l11 in stringent control of rna synthesis in bacillus megaterium.a spontaneously occurring thiostrepton-resistant mutant of bacillus megaterium has been shown to yield ribosomes lacking protein bm-l11, a protein immunologically related to escherichia coli ribosomal protein l11. here we have demonstrated that the mutant strain has acquired the relaxed phenotype and is unable to synthesise guanosine tetraphosphate and pentaphosphate in vivo. ribosomes from the mutant strain are unable to support the synthesis of these two compounds in vitro, but this deficiency ...1979118006
dielectric properties of native and decoated spores of bacillus megaterium.a general model for use in interpreting dielectric data obtained with bacterial endospores is developed and applied to past results for bacillus cereus spores and new results for bacillus megaterium spores. the latter were also subjected to a decoating treatment to yield dormant cells with damaged outer membranes that could be germinated with lysozyme. for both spore types, core ions appeared to be completely immobilized, and decoating of b. megaterium spores did not affect this extreme state of ...1979118161
energetics of rapid transmembrane movement and of compositional asymmetry of phosphatidylethanolamine in membranes of bacillus megaterium.the energy requirements for the rapid transmembrane movement of phosphatidylethanolamine in membranes of bacillus megaterium km have been investigated by means of pulse label experiments. the transmembrane movement continues at a high rate in cells blocked in the production of metabolic energy by treatment with a combination of inhibitors. the movement is shown to be completely independent of the synthesis of lipid and of protein and, more generally, independent of sources of metabolic energy. t ...1979118463
ribosomes in thiostrepton-resistant mutants of bacillus megaterium lacking a single 50 s subunit protein. 1979119865
on the biological role of ribosomal protein bm-l11 of bacillus megaterium, homologous with escherichia coli ribosomal protein l11. 1979119869
influence of potassium chloride upon the binding and antibacterial activity of chlorhesidine diacetate and (ethoxy)5 octyl phenol (triton x45) towards bacillus megaterium km-.chlorhexidine (0.5-0.65 microm) and triton x45 (30-40 microm) added to exponential phase bacillus megaterium km- cultures was growth inhibitory. the presence of kcl (0.05-0.35 m) in the medium did not significantly affect growth rate in the absence of drug, yet reduced the growth inhibitory activity of the chlorhexidine and enhanced that of triton x45. these effects were maximal at kcl concentrations of 0.2 m and above, when complete protection towards chlorhexidine and lysis of the cultures in ...1979120929
structure of the surface of spores and morphological-physiological properties of individual strains of bacillus megaterium.the structure of the surface of spores of 10 strains of bacillus megaterium was investigated by the method of carbon replicas. they were separated into three groups according to the peculiarities of the structural organization of the surface of the spores. the combination of other morphological and physiological-biochemical characteristics of these strains was characteristic of the species bac. megaterium. strains with differences in the structure of the spore surface also showed similarities in ...1979121542
bacillus megaterium resistance to cloxacillin accompanied by a compensatory change in penicillin binding proteins. 1979121895
morphogenesis of the membrane-bound electron-transport system in sporulating bacillus megaterium km.the properties of electron transport systems present in soluble and particulate fractions of spores of bacillus megaterium km?have been compared with those of similar fractions prepared from exponential-phase vegetative cells of this organism. the timing and localization of modifications of the electron transport system occurring during sporulation have been investigated by using a system for separating forespores from mother cells at all stages during development [8]. spore membranes contained ...1975126154
biochemical evidence for the reversed polarity of the outer membrane of the bacterial forespore.measurement of certain membrane-bound enzymic activities was used to study the orientation of the outer membrane of the double-membraned forespore of bacillus megaterium km. 2. adenosine triphosphatase, nadh dehydrogenase and l-malate intact protoplasts, but were readily detected in intact stage ii or iv forespores, consistent with reversed polarity of the outer forespore membrane relative to the mother-cell plasma membrane. 3. measurement of nadh oxidase activity revealed that intact stage iii ...1975132169
the effect of atebrin on bacterial membrane adenosine triphosphatases in relation to the divalent cation used as substrate and/or activator.the action of atebrin on purified adenosine triphosphatase (atpase) from micrococcus lysodeikticus was studied as well as on the membrane-bound and soluble atpases from escherichia coli and bacillus megaterium. atebrin inhibited the ca(2+)-dependent activity of all these enzymes, and the inhibition was reversed by an excess of ca(2+) ions. kinetic studies carried out with the purified enzyme from m. lysodeikticus showed that the inhibition by atebrin was strongly cooperative, suggesting the comp ...1977138384
bacillus megaterium mutant deficient in membrane-bound adenosine triphosphatase adenosine triphosphatase (atpase) mutant of bacillus megaterium was isolated and characterized. this mutant (designated a37) was unable to grow on nonfermentable carbon sources and possessed less than 5% of the wild-type atpase activity. oxygen uptake by the mutant was comparable to that in the wild type. sporulation in the wild type occurred in both glucose- and nitrogen-limiting media; however, a37 sporulated only in the nitrogen-limiting medium. the inability of a37 to sporulate in glucose ...1977141449
membrane changes during germination of bacillus megaterium km spores.bacillus megaterium km dormant spore inner membrane atpase exhibits a ten-fold increase in specific activity during the first 10 min of germination in the absence of protein synthesis. during this time period both in the presence and absence of chloramphenicol extensive proteolysis of spore inner membrane takes place, which results in degradation of approximately half of the membrane protein. polyacrylamide gel electrophoresis reveals the extent and selectivity of this degradation of inner membr ...1979161797
characterization of a cytochrome p-450-dependent steroid hydroxylase system present in bacillus megaterium.cell-free extracts from sonically disrupted bacillus megaterium atcc 13368 hydroxylated a variety of 3-oxo-delta4-steroids in position 15beta in the presence of nadph and o2. ring a-reduced, aromatic and 3beta-hydroxy-delta5-steroids did not serve as substrates for the 15beta-hydroxylase system. using ion exchange chromatography on deae-cellulose and gel filtration on ultrogel aca-54 it was possible to resolve the hydroxylase system into three proteins: a strictly nadph-dependent fmn-containing ...1976177422
evidence against the involvement of adenosine 3',5'-cyclic monophosphate in glucose inhibition of beta-galactosidase induction in bacillus megaterium.when bacillus megaterium cells are grown on d-galactose as the sole carbon source, the cells actively synthesize beta-galactosidase (beta-d-galactoside galactohydrolase, ec however, d-galactose, when added to a glucose-grown culture, did not induce beta-galactosidase, apparently because of the glucose inhibition of the transport of galactose. on the other hand, when glucose was added to a galactose-grown culture, the transport of galactose continued at a reduced but significate rate, ...1976186165
levels of small molecules and enzymes in the mother cell compartment and the forespore of sporulating bacillus megaterium.we have determined the amounts of a number of small molecules and enzymes in the mother cell compartment and the developing forespore during sporulation of bacillus megaterium. significant amounts of adenosine 5'-triphosphate and reduced nicotinamide adenine dinucleotide were present in the forespore compartment before accumulation of dipicolinic acid (dpa), but these compounds disappeared as dpa was accumulated. 3-phosphoglyceric acid (3-pga) accumulated only within the developing forespore, be ...1977193830
production of large amounts of acetate during germination of bacillus megaterium spores in the absence of exogenous carbon sources.when bacillus megaterium spores germinate in the absence of an exogenous carbon source, the first minutes of germination are accompanied by production of large amounts (approximately 70 nmol/mg of dry spores) of acetate and much smaller amounts of pyruvate and lactate. the majority of these compounds are excreted into the medium. exogenous pyruvate and alanine are also converted to co2 and acetate by germinating spores, presumably by using the pyruvate dehydrogenase that is present in dormant sp ...1977199580
the purification and characterization of a dna nicking-closing enzyme from bacillus megaterium.although several eucaryote dna nicking--closing (n--c) enzymes have been characterized, only the escherichia coli enzyme has been extensively studied amongst procaryotes. the latter enzyme is distinctly different from the eucaryotic enzymes and we have therefore purified the n--c enzyme from bacillus megaterium to determine if procaryotes form a distinctive class of n--c enzymes. the purified b. megaterium n--c enzyme has a molecular weight of 120,000, only partly relaxes negative supercoils, do ...1978204401
synthesis and quantitative structure-activity relationships of antibacterial 1-(substituted benzhydryl)-4-(5-nitro-2-furfurylideneamino) piperazines.1-benzhydryl -4- (5-nitro-2-furfurylideneamino) piperazine and 11 substituted analogs were prepared and examined for in vitro antimicrobial activity. the compounds were active against bacillus cereus 7, bacillus megaterium 122, bacillus subtilis 104, clostridium perfringens 13, and the tetracycline-resistant clostridium perfringens 37. regression analyses on the antibacterial activity data based on the hansch approach, using pi, pi2, and sigma parameters, yielded several statistically significan ...1978207855
phosphoglycerate mutase in developing forespores of bacillus megaterium may be regulated by the intrasporal level of free manganous ion. 1978208551
levels of cyclic gmp in dormant, germinated, and outgrowing spores and growing and sporulating cells of bacillus megaterium.the level of cyclic gmp was less than one molecule per organism in dormant, germinated, and outgrowing spores of bacillus megaterium. a significant level (approximately 8 pmol/g, dry weight) of cyclic gmp was found in early to mid-log phase cells, but the level fell to below 0.2 pmol/g, dry weight, in late-log phase and only rose slightly to approximately 0.9 pmol/g, dry weight, in stationary phare. no significant amount of cyclic gmp was detected in the growth medium at any time.1978213418
influence of suspending media upon the susceptibility of pseudomonas aeruginosa nctc 6750 and its spheroplasts to polymyxin b.the lytic and bactericidal actions of polymyxin b on whole cells and spheroplasts of pseudomonas aeruginosa varied markedly with the suspending media, and there was little correlation between them. relative rates of lysis of these preparations and also of bacillus megaterium protoplasts suggested that polymyxin causes progressive damage to the cytoplasmic membrane, such that membrane permeability towards various ions increased as follows: k(+) > na(+) > no(3) (-) > cl(-), ca(2+), h(2)po(4) (-)/h ...1979218501
regulation of phosphoglycerate phosphomutase in developing forespores and dormant and germinated spores of bacillus megaterium by the level of free manganous ions.the large depot of phosphoglyceric acid (pga) which is accumulated within spores of bacillus megaterium is greater than 99% 3-phosphoglyceric acid (3-pga). the 3-pga depot is stable in forespores and dormant spores, but is utilized rapidly during spore germination. when spores were germinated in kbr plus naf, the pga depot was not utilized, but 13% of the 3-pga was converted to 2-pga. these data suggest phosphoglycerate phosphomutase as the enzyme which is regulated to allow 3-pga accumulation d ...1979225303
the role of manganese in growth and sporulation of bacillus subtilis.phosphoglycerate phosphomutase of bacillus subtilis, bacillus cereus and bacillus megaterium required mn2+ as cofactor, whereas the wheat germ and rabbit liver enzymes did not. in the absence of mn2+, b. subtilis did not sporulate in normal sporulation media but it did sporulate if the proper ratio of glucose or glycerol and malate was used. decoyinine, an inhibitor of guanosine monophosphate synthesis, induced sporulation in the presence of excess glucose and malate to the same extent with and ...1979225409
use of electron spin resonance to study bacillus megaterium spore membranes. 1979226087
isolation and characterization of cytochrome p-450meg.the cytochrome p-450-dependent steroid 15 beta-hydroxylase system from bacillus megaterium has been resolved into three components, 1) a nadph-specific, fmn-containing flavoprotein reductase, molecular weight 55-60 000; 2) an iron-sulfur protein, molecular weight 13,000 and 3) cytochrome p-450meg, molecular weight 52,000. the cytochrome component has been purified to homogeneity, as judged by sds-polyacrylamide gel electrophoresis and isoelectric focusing in polyacrylamide gel, and its amino aci ...1979229681
biosynthesis of phospholipids in bacillus megaterium.information on the biosynthesis of phospholipids in bacteria has been derived principally from the study of escherichia coli and other gram-negative organisms. we have now carried out a detailed study of the pathways of phospholipid biosynthesis in the gram-positive organism bacillus megarterium km in relation to investigations on the biogenesis of lipid asymmetry in membranes. radioactive precursors such as 32pi and [3h]palmitate initially label phosphatidylethanolamine much more than phosphati ...1979230180
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