| [renaturation of bacterial metalloproteinases]. | after denaturation by phenol or acid ethanol bacterial metalloproteinases secreted by bacillus thermoproteolyticus and bacillus megaterium cells can be renatured by dissolution in 99.7% formic acid with subsequent dilution of the solution and its neutralization with an appropriate amount of an alkali. renaturation is optimal at ph 9.0 in the presence of 30% glycerol as stabilizer, ca2+ and zn2+ ions needed for the formation of a native structure and reconstitution of the enzyme catalytic center. ... | 1993 | 8364114 |
| distribution and identification of proteolytic bacillus spp. in paddy field soil under rice cultivation. | proteolytic bacteria in paddy field soils under rice cultivation were characterized and enumerated using azocoll agar plates. bacillus spp. were the proteolytic bacteria that were most frequently present, comprising 59% of the isolates. they were always the numerically dominant proteolytic bacteria isolated from three kinds of fertilizer treatments (yearly application of rice-straw compost and chemical fertilizer, yearly application of chemical fertilizer, and no fertilizer application) and at t ... | 1993 | 8364803 |
| isolation of three antibacterial peptides from pig intestine: gastric inhibitory polypeptide (7-42), diazepam-binding inhibitor (32-86) and a novel factor, peptide 3910. | two antibacterial peptides, cecropin p1 and pr-39 (39-residue proline/arginine-rich peptide), from the upper part of pig small intestine have previously been isolated and characterized. we have now continued our search for antibacterial peptides in different side fractions generated during the isolation of intestinal hormones. starting from one such fraction and monitoring activity against bacillus megaterium, we isolated three homogeneous peptides by three consecutive chromatographic steps. ami ... | 1993 | 8375398 |
| identification of charge-transfer transitions in the optical spectrum of low-spin ferric cytochrome p-450 bacillus megaterium. | the optical, low temperature magnetic circular dichroism (mcd) and epr spectra of low-spin fe(iii) cytochrome p-450 bm-3 from bacillus megaterium, and its imidazole adduct have been measured. the mcd spectra locate new transitions over 600-700 nm and 800-1300 nm. the latter are assigned to porphyrin (a1u)-fe(iii) (dyz) charge-transfer (ct) transitions. in the case of the imidazole adduct the energy of this transition fits well to the theoretical model of gadsby and thomson [gadsby, p. m. a. & th ... | 1993 | 8386633 |
| netropsin inhibits the increase of intracellular ca(2+)-dependent serine proteinase activity in sporulating bacillus megaterium. | netropsin suppressed the increase of intracellular proteolytic activity when added to b. megaterium incubated in a sporulation medium. the inhibited enzyme was a ca(2+)-dependent serine proteinase. sporulation and protein turnover in later sporulation phases were inhibited as well. different concentrations of netropsin affected various aspects of protein catabolism differently. | 1993 | 8388845 |
| cloning and nucleotide sequence of a plasmid-carried gene coding for a minor small, acid-soluble protein from bacillus megaterium spores. | the gene (termed sspg) coding for a small, acid-soluble protein (sasp) from spores of bacillus megaterium qmb1551, termed sasp-g, has been cloned, and its nucleotide sequence has been determined. sasp-g is a 42-residue protein containing 2 tryptophan and 11 lysine residues, including a hexalysine sequence, and is not homologous to any previously described sasp. the sspg gene appears to be an additional member of the sigma g regulon. no gene homologous to sspg is present in b. cereus t or b. subt ... | 1993 | 8407806 |
| cell wall assembly in staphylococcus aureus: proposed absence of secondary crosslinking reactions. | the distribution of muropeptides formed by muramidase digestion of peptidoglycan from staphylococcus aureus h was determined by gel-filtration hplc. the observed crosslinking pattern supports the conclusion that incorporation of peptidoglycan in s. aureus proceeds by a similar mechanism to that proposed earlier for bacillus megaterium. in this mechanism single glycan-peptide strands are incorporated into the sacculus by crosslinking reactions that take place only between the monomer muropeptide ... | 1993 | 8409927 |
| complementation of an enterococcus hirae (streptococcus faecalis) mutant in the alpha subunit of the h(+)-atpase by cloned genes from the same and different species. | we isolated an enterococcus hirae (formerly streptococcus faecalis) mutant, designated ms117, in which 'g' at position 301 of the alpha-subunit gene of the f1f0 type of h(+)-atpase was deleted. ms117 had low h(+)-atpase activity, was deficient in the regulatory system of cytoplasmic ph, and was unable to grow at ph 6.0. when the alpha-subunit gene of e. hirae h(+)-atpase was ligated with the shuttle vector phy300plk at the downstream region of the tet gene of the vector, it was expressed without ... | 1993 | 8412656 |
| mutations that significantly change the stability, flexibility and quaternary structure of the l-lactate dehydrogenase from bacillus megaterium. | in order to investigate the physical basis of protein stability, two mutant l-lactate dehydrogenases (ldh) and the wild-type enzyme from bacillus megaterium were analyzed for differences in quaternary structure, global protein conformation, thermal stability, stability against guanidine hydrochloride, and polypeptide chain flexibility. one mutant enzyme, ([t29a, s39a]ldh), differing at two positions in the alpha-b helix, exhibited a 20 degrees c increase in thermostability. hydrogen/deuterium ex ... | 1993 | 8425537 |
| inhibition by barbiturates of the binding of bm3r1 repressor to its operator site on the barbiturate-inducible cytochrome p450bm-3 gene of bacillus megaterium. | in our previous publication (shaw, g.-c., and fulco, a. j. (1992) j. biol. chem. 267, 5515-5526), we reported that bm3r1, a protein encoded in an open reading frame just upstream from the cytochrome p450bm-3 gene, is a repressor critically involved in the barbiturate-inducible expression of this gene in bacillus megaterium. we now describe the purification of the bm3r1 protein from an overproducing escherichia coli strain harboring a bm3r1 gene-carrying plasmid and report the effect of barbitura ... | 1993 | 8428974 |
| cloning, sequencing and expression of the gene encoding glucose dehydrogenase from the thermophilic archaeon thermoplasma acidophilum. | the gene encoding glucose dehydrogenase has been identified by southern analysis of doubly restricted genomic thermoplasma acidophilum dna, using two redundant 17-residue oligonucleotide probes reverse translated from protein n-terminal sequence data. a 1670-bp bamh1-ecor1 restriction fragment was ligated into puc19 and puc18 (constructs ptagdh1 and ptagdh2, respectively) and cloned in escherichia coli. the sequence of the whole fragment was determined, and a 1059-bp open reading frame identifie ... | 1993 | 8436115 |
| molecular cloning and nucleotide sequence of the gene encoding a calcium-dependent exoproteinase from bacillus megaterium atcc 14581. | the gene nprm encoding the calcium-dependent extracellular proteinase from bacillus megaterium atcc 14581 was cloned in the vector pbr322 and expressed in escherichia coli hb101. the dna sequence of the cloned 3.7 kb fragment revealed only one open reading frame consisting of 1686 bp with a coding capacity of 562 amino acid residues. a predicted shine-dalgarno (sd) sequence was observed 9 bp upstream from the presumptive translation start site (atg). a possible promoter sequence (tagacg for the ... | 1993 | 8450307 |
| critical residues involved in fmn binding and catalytic activity in cytochrome p450bm-3. | cytochrome p450bm-3 from bacillus megaterium is a soluble, catalytically self-sufficient fatty acid mono-oxygenase that, in structural organization and amino acid sequence, resembles the class ii (microsomal) p450 systems. its single polypeptide chain contains both a p450 heme domain and an nadph:p450 reductase domain, each of which bears significant homology with its microsomal counterparts. we report here the critical nature of three amino acids in the reductase domain of this enzyme with resp ... | 1993 | 8463285 |
| proteolytic processing of the protease which initiates degradation of small, acid-soluble proteins during germination of bacillus subtilis spores. | degradation of small, acid-soluble spore proteins during germination of bacillus subtilis spores is initiated by a sequence-specific protease called gpr. western blot (immunoblot) analysis of either bacillus megaterium or b. subtilis gpr expressed in b. subtilis showed that gpr is synthesized at about the third hour of sporulation in a precursor form and is processed to an approximately 2- to 5-kda-smaller species 2 to 3 h later, at or slightly before the time of accumulation of dipicolinic acid ... | 1993 | 8478323 |
| the cortical peptidoglycan from spores of bacillus megaterium and bacillus subtilis is not highly cross-linked. | determination by amino acid analyses of the percentage of diaminopimelic acid in the spore cortex of bacillus megaterium and bacillus subtilis which is involved in interpeptide cross-links gave values of 31 to 37%. this finding supports the idea that the cortex volume could undergo significant changes in response to changes in ph or ionic strength and could thus play an active role in reducing the water content of the spore protoplast during sporulation. | 1993 | 8478339 |
| [the use of the neustonic forms of bacilli for purifying and decontaminating reservoirs]. | it is shown possible to select the bacterial strains which are neuston ones, i.e., concentrating on the water-atmosphere interface. the preparation based on the neuston form of bacillus megaterium is more efficient for purification of water polluted with oil hydrocarbons than the preparation based on the planktonic form of the same culture. preparation based on the neuston form of the aerobic spore-forming bacteria is effective for biological decontamination of sewage treated using conventional ... | 1993 | 8497205 |
| purification, characterization, and lytic activity against naegleria fowleri of two amoebicins produced by bacillus licheniformis a12. | bacillus licheniformis a12 produces two amoebolytic substances (amoebicins a12-a and a12-b) in liquid media during sporulation. both substances have been purified and characterized. they are heat- and protease-resistant peptides containing aspartic acid, glutamic acid, serine, proline, and tyrosine in a molar ratio of 5:2:2:2:2. no fatty acids or carbohydrates have been detected. their molecular weight is 1,430. purified amoebicins a12-a and a12-b exhibit amoebolytic action against naegleria fow ... | 1993 | 8517742 |
| close evolutionary relatedness among functionally distantly related members of the (alpha/beta)8-barrel glycosyl hydrolases suggested by the similarity of their fifth conserved sequence region. | a short conserved sequence equivalent to the fifth conserved sequence region of alpha-amylases (173_lpdld, aspergillus oryzae alpha-amylase) comprising the calcium-ligand aspartate, asp-175, was identified in the amino acid sequences of several members of the family of (alpha/beta)8-barrel glycosyl hydrolases. despite the fact that the aspartate is not invariantly conserved, the stretch can be easily recognised in all sequences to be positioned 26-28 amino acid residues in front of the well-know ... | 1995 | 8543020 |
| the structure of bacterial cell cycle and age structure of bacterial populations. | study of synchronous and asynchronous cultures of bacillus megaterium, bacillus thuringiensis and bacillus licheniformis has shown that the duration of chromosomal dna replication (period c) is proportional to the generation time, and time between two cycles of the dna replication (known as period i). the duration of period c is nearly constant and makes up from 0.5 to 1.0 hour at the variations of the generation time from 1.5 to 2.75 hours. the duration of period b (the time between the termina ... | 1995 | 8548068 |
| bacteriolytic activities of the free-living soil amoebae, acanthamoeba castellanii, acanthamoeba polyphaga and hartmannella vermiformis. | bacteriolytic activities of axenically grown free-living soil amoebae acanthamoeba castellanii, acanthamoeba polyphaga and hartmannella vermiformis towards various gram-positive and gram-negative bacteria were determined. a spectrophotometric assay revealed that the specific bacteriolytic activities of both acanthamoeba species were higher as those of the three hartmannella strains. bacillus megaterium, bacillus subtilis, chromatium vinosum, micrococcus luteus and pseudomonas fluorescens were mo ... | 1995 | 8572682 |
| adventures with bacillus megaterium--fusion of bacterial protoplasts. | this essay describes the author's studies with bacteria in post-war hungary; the difficulties encountered, with funding, collaboration and publication; and how the szeged institute consolidated itself as the one outstanding scientific institute in eastern europe, with the author at the helm. | 1996 | 8593168 |
| occurrence of polyhydroxyalkanoic acid granule-associated proteins related to the alcaligenes eutrophus h16 ga24 protein in other bacteria. | fifty different polyhydroxyalkanoic acid (pha)-accumulating bacterial strains were investigated for the occurrence of phasin proteins bound to pha granules and related to the ga24 protein of alcaligenes eutrophus h16, by isolating pha granules and western blot analysis of granule-associated proteins employing antibodies raised against the ga24 protein. it could be demonstrated that th pha granules of many poly(3-hydroxybutyrate)-accumulating bacteria exhibited a similar protein pattern, and a pr ... | 1996 | 8598273 |
| contributions of xylr ccpa and cre to diauxic growth of bacillus megaterium and to xylose isomerase expression in the presence of glucose and xylose. | bacillus megaterium shows diauxic growth in minimal medium containing glucose and xylose. we have examined the influence of three elements that regulate xyl operon expression on diauxic growth and expression of a xyla-lacz fusion. xyla is 13-fold repressed during growth on glucose. induction occurs at the onset of the lag phase after glucose is consumed. inactivation of xylr yields a two-fold increase in expression of xyla on glucose. deletion of the catabolite responsive element (cre) has a mor ... | 1996 | 8602140 |
| biochemical and antibacterial analysis of human wound and blister fluid. | fluid from a post-operative wound, six leg ulcers and a large blister were collected and analysed by biochemical, microbiological and immunological techniques. the results were compared with those from sera. all samples were lyophilized and extracted twice with 60% aqueous acetonitrile containing 1% trifluoroacetic acid. the pooled supernatants were lyophilized, redissolved, and the fluid extracts were characterized by six techniques (the blister exudate only with three): reverse-phase hplc, edm ... | 1996 | 8620898 |
| induction of cytochrome p-450 bm-3 (cyp 102) by non-steroidal anti-inflammatory drugs in bacillus megaterium. | bacillus megaterium contains a cytochrome p-450 fatty acid mono-oxygenase which is inducible with barbiturate drugs. we have demonstrated that this enzyme system is inducible with peroxisome proliferators. in mammals, peroxisome proliferators also induce mono-oxygenases in the cyp4a gene family. in this paper we demonstrate that the non-steroidal anti-inflammatory drugs ibuprofen, ketoprofen and indomethacin are potent inducers of fatty acid mono-oxygenase activity as well as of p-450bm-3 protei ... | 1996 | 8645218 |
| the c-terminal bisphosphorylated proenkephalin-a-(209-237)-peptide from adrenal medullary chromaffin granules possesses antibacterial activity. | the chromaffin granules have been shown to be an excellent model to study the processing of proenkephalin-a and chromogranins. recently, we reported a study dealing with the processing of chromogranin b/secretogranin i and the occurrence of the c-terminal chromogranin b-derived peptide 614-626 which was shown to have antibacterial activity [strub, j.m., garcia-sablone, p., looning, k., taupenot, l., hubert, p., van dorsselaer, a., aunis, d. & metz-boutigue, m.h. (1995) eur. j. biochem. 229, 356- ... | 1996 | 8654396 |
| characterization of the major citrate synthase of bacillus subtilis. | the major citrate synthase of bacillus subtilis (cs-ii) was purified to near homogeneity and shown to correspond to the product of the citz gene. accumulation of cs-ii during exponential growth and stationary phases paralleled expression of the citz gene. the physical and kinetic properties of cs-ii were similar to those of citrate synthase enzymes from bacillus megaterium and from eukaryotic cells but differed from those of citrate synthases from many gram-negative bacteria. | 1996 | 8655569 |
| immunological crossreactivity to the catabolite control protein ccpa bacillus megaterium is found in many gram-positive bacteria. | the catabolite control protein ccpa from bacillus megaterium was overproduced as a fusion protein to a 6xhis affinity tag and purified to homogeneity. polyclonal antibodies of high affinity and specificity were raised against the purified protein. the serum did not crossreact with purified lac repressor despite the fact that ccpa and laci belong to the same protein family. using this antiserum we identified proteins that share antigenic determinants with ccpa in many gram-positive bacteria, incl ... | 1996 | 8674978 |
| [interspecific protoplast fusion between bacillus thuringensis bt-3701 and bacillus megaterium bm-107]. | the results of the interspecific protoplast fusion between b. thuringensis sub. kurstaki bt-3701 which has pesticide ability, and b. megaterium var. phosphaticum bm-107 which has decomposing phosphate activity, were reported. high frequency of protoplast formation and regeneration was obtained with 4h activated bm-107 treated by 100 micrograms/ml lysozyme, and with 2h activated bt-3701 treated by 3% glycin and mild temperature. using 40% peg and 5% nascent ca2+ to treat the parential protoplast ... | 1995 | 8701582 |
| expression of periplasmic alpha-amylase of xanthomonas campestris k-11151 in escherichia coli and its action on maltose. | a gene encoding the periplasmic alpha-amylase of xanthomonas campestris k-11151 was cloned into escherichia coli using puc19 as a vector. an orf of 1578 bp was deduced to be the amylase structural gene. the primary structure of the enzyme had little identity with other alpha-amylases, except with the enzyme from bacillus megaterium. the enzyme was expressed in e. coli from the lac promoter of puc19 and was found to be transported to the periplasmic space. the expressed enzyme showed the same the ... | 1996 | 8704990 |
| probing electron transfer in flavocytochrome p-450 bm3 and its component domains. | rapid events in the processes of electron transfer and substrate binding to cytochrome p-450 bm3 from bacillus megaterium and its constituent haem-containing and flavin-containing domains have been investigated using stopped-flow spectrophotometry. the formation of a blue semiquinone flavin form occurs during the nadph-dependent reduction of the flavin domain and a species with a similar absorption maximum is also seen during reduction of the holoenzyme by nadph. epr spectroscopy confirms the fo ... | 1996 | 8706747 |
| purification and characterization of 6xhis-tagged bm3r1 repressor of bacillus megaterium. | the bm3r1 gene-encoded repressor controls the expression of cytochrome p450bm-3 gene as well as its own expression in bacillus megaterium. we have developed an efficient system for overexpression and purification of bm3r1 protein by nickel ion affinity chromatography. adding six histidine residues at either n-terminus or c-terminus of bm3r1 repressor caused no loss of its operator dna binding ability. we have investigated the interaction between bm3r1(his)6 and its operator dna by equilibrium an ... | 1995 | 8747550 |
| cytochromes p450 of the sophorose lipid-producing yeast candida apicola: heterogeneity and polymerase chain reaction-mediated cloning of two genes. | candida apicola belongs to a group of yeasts producing high amounts of surface-active extracellular glycolipids consisting of sophorose and long-chain-omega- and (omega-1)-hydroxy fatty acids. the involvement of cytochrome p450 in the synthesis of sophorose lipid by the hydroxylation of long-chain fatty acids was suggested from a simultaneous increase of cellular p450 content. hydroxylation studies indicated the existence of multiple p450 forms capable of hydroxylating not only long-chain fatty ... | 1996 | 8771711 |
| sporicidal action of peracetic acid and protective effects of transition metal ions. | although peracetic acid (paa) is used widely for cold sterilization and disinfection, its mechanisms of sporicidal action are poorly understood. paa at high concentrations (5-10%) can cause major loss of optical absorbance and microscopically-visible damage to bacterial spores. spores killed by lower levels of paa (0.02-0.05%) showed no visible damage and remained refractile. treatment of spores of bacillus megaterium atcc 19213 with paa at concentrations close to the lethal level sensitized the ... | 1995 | 8821509 |
| the aggregation-mediated conjugation system of bacillus thuringiensis subsp. israelensis: host range and kinetics of transfer. | the aggregation-mediated conjugation system in bacillus thuringiensis subsp. israelensis encoded on the plasmid pxo16 is characterized by the formation of aggregates when agr+ and agr- cells are socialized in exponential growth. using the aggregation phenotypes, we have identified potential recipients of the aggregation-plasmid pxo16 among bacillus cereus, bacillus subtilis, bacillus megaterium, bacillus sphaericus, and 24 subspecies of b. thuringiensis. we found 14 agr- strains, i.e., potential ... | 1996 | 8824168 |
| cytochrome p450foxy, a catalytically self-sufficient fatty acid hydroxylase of the fungus fusarium oxysporum. | we have purified membrane-bound fatty acid (omega-1-omega-3) hydroxylase of the fungus fusarium oxysporum mt-811 and found that the activity depends on a single polypeptide with an apparent m(r) value of 118,000. the purified hydroxylase exhibited spectral characteristics of cytochrome p450 (p450), and could catalyze the hydroxylation without the aid of any other proteinaceous components, such as nadph-p450 reductase. these properties of the fungal hydroxylase are the same as those of bacterial ... | 1996 | 8830036 |
| structure and function of the bacillus spoiie protein and its localization to sites of sporulation septum assembly. | functioning of the spoiie locus of bacillus subtilis is required for formation of a normal polar septum during sporulation and for activation of the transcription factor sigma f, which directs early forespore-specific gene expression. we have determined the dna sequence of the wild type and several mutant alleles of the spoiie gene of b. subtilis and sequenced a substantial portion of its presumptive homologue in bacillus megaterium. we show that the spoiie locus encodes a single large protein w ... | 1996 | 8830262 |
| induction and carbon catabolite repression in the biosynthesis of beta-amylase by bacillus megaterium b6. | biosynthesis of extracellular beta-amylase in bacillus megaterium b6 was induced by starch, although maltodextrin was found to be the actual inducer. amongst the carbon sources tested, glucose was found to be the most potent repressor and when added exogenously to a starch-induced culture it brought about an immediate fall in enzyme synthesis through carbon catabolite repression. the repression was not overcome after the fall of glucose concentration in the culture medium below a critical level. ... | 1996 | 8850517 |
| cloning and sequencing of the spore germination gene of bacillus megaterium atcc 12872: similarities to the nah-antiporter gene of enterococcus hirae. | the germination mutant tm-31 of bacillus megaterium atcc 12872, was isolated by transposon tn917 insertional mutagenesis. glucose, l-proline, l-leucine and kno3 germinated tm-31 poorly. the dna in the region of the tn917 insertion was cloned, and its nucleotide sequence determined. one major open reading frame was present on the cloned dna. the hydrophobic protein encoded is presumably membrane-associated. a homology search revealed that the gene encoded in the region of the tn917 insertion is h ... | 1996 | 8867604 |
| asporogenic bacillus megaterium mutant 27-36 degrades intrinsically short-lived proteins but fails to convert most of other proteins to a short-lived fraction. | asporogenic mutant blocked in the 0-ii sporulation stage degraded pulse-labelled proteins in the sporulation medium at the same rate as the parental strain for the first two hours. the degraded fraction was mostly composed of intrinsically short-lived proteins which were degraded even after enriching the medium with amino acids and growth resumption. proteins accessible to degradation because of nutritional shift down formed a lesser proportion of this fraction. the acceleration of protein turno ... | 1996 | 8876972 |
| catabolite repression mediated by the catabolite control protein ccpa in staphylococcus xylosus. | the gene ccpa encoding the catabolite control protein ccpa of staphylococcus xylosus has been cloned and characterized. the ccpa protein belongs to the lacl/gair family of bacterial regulators and is comprised of 329 amino acids, with a molecular mass of 36.3 kda. it shows 56% identity with the ccpa proteins of bacillus subtills and bacillus megaterium. inactivation of the ccpa gene in the genome of s. xylosus relieved the activities of three enzymes, alpha-glucosidase, beta-glucuronidase, and b ... | 1996 | 8878037 |
| bacterial contaminants in liquid packaging boards: assessment of potential for food spoilage. | liquid packaging boards and blanks were examined for microbial contaminants. a total of 218 strains were identified and representatives of the most frequent species were characterized for their potential for food spoilage. contaminants found were aerobic spore-forming bacteria, mostly bacillus megaterium, b. licheniformis, b. cereus group, b. pumilus, paenibacillus macerans, p. polymyxa, p. pabuli and b. flexus. production of amylolytic, proteolytic, lipolytic and phospholipolytic enzymes was co ... | 1996 | 8896355 |
| activation of the intracellular ca(2+)-dependent serine protease isp1 of bacillus megaterium by purification or by high ca2+ concentrations. | the total proteolytic activity of isp1 determined after its partial purification by size exclusion hplc increased 3.0, 7.3 and 27.3 times in sporulating, growing and netropsin-treated cells, respectively, as compared with the corresponding original activity in crude cytoplasmic preparations at the same cacl2 concentration of 3 mm. a similar rise in proteolytic activity occurred on increasing the ca2+ concentration in the crude cytoplasm from 3 to 30 mm. this activation in the cytoplasm of netrop ... | 1996 | 8955884 |
| localization of a conformational energy-coupling determinant near the c terminus of the beta subunit of the f1f0-atpase. | escherichia coli mutants in the beta subunit of the f1f0-atpase can be complemented with the beta subunit from the obligate aerobe bacillus megaterium. it has been shown that cells carrying such hybrid atpases have an unusual energy-coupling phenotype. although they are able to grow on minimal succinate medium, and therefore carry a functional atp synthase, they are defective in the ability to grow anaerobically, indicating some defect in atp-driven proton pumping (scarpetta, m., hawthorne, c. a ... | 1996 | 8969200 |
| a xylose-inducible bacillus subtilis integration vector and its application. | the construction of a xylose-inducible expression vector is described. this vector allows the integration of any gene, coding for its authentic protein, at the amye locus of bacillus subtilis (bs). the controlable expression cassette consists of the repressor-encoding gene and the promoter of the bacillus megaterium-derived operon for xylose utilization, sandwiched between the 5'- and 3'-ends of amye. this thereby allows insertion of in vitro constructed transcriptional fusions at the amye locus ... | 1996 | 8973310 |
| a 53-base-pair inverted repeat negatively regulates expression of the adjacent and divergently oriented cytochrome p450(bm-1) gene and its regulatory gene, bm1p1, in bacillus megaterium. | to study the role of the cis-acting element(s) in controlling the expression of the cytochrome p450(bm-1) gene and its upstream regulatory gene, bm1p1, in bacillus megaterium, various deletion derivatives were constructed. a 53-bp inverted repeat located midway between the p450(bm-1) gene and bm1p1 gene was found in vivo to negatively regulate the expression of both genes, the regulation of which may occur at the transcriptional level. the promoter of the p450(bm-1), gene was also identified and ... | 1997 | 8982010 |
| nk-lysin, structure and function of a novel effector molecule of porcine t and nk cells. | nk-lysin (nkl), a 78-residue antimicrobial peptide, was isolated from pig small intestine. standard methods identified the peptide as basic, with six half-cystine residues in three intrachain disulphide bonds. the sequence showed 33% identity with a part of a putative gene product (nkg5) from activated t and nk cells, nk-lysin showed antibacterial activity against escherichia coli and bacillus megaterium and marked lytic activity against yac-1, a nk sensitive tumour cell line, while sheep red bl ... | 1996 | 8988855 |
| pr-39, a proline-rich peptide antibiotic from pig, and fall-39, a tentative human counterpart. | the peptide antibiotic pr-39 was originally isolated from the upper part of pig intestine. it has antibacterial activity against gram negative bacteria at concentrations comparable with tetracycline. studies of the mechanism of action showed that pr-39 inhibits both dna and protein synthesis. recently, pr-39 was found in wound fluid and was shown to have inductive activity on matrix components as part of the wound repair process. we have now sequenced the complete gene and possible mediators of ... | 1996 | 8988856 |
| phenobarbital-dependent protein binding to barbie box-like sequences in the coding region of cytochrome p450bm-3 gene from bacillus megaterium. | phenobarbital-dependent protein binding was shown to occur to dna fragments from the coding region of the cytochrome p450bm-3 gene from bacillus megaterium. incubation of the dna fragments from the coding region of the gene with total cell extract from bacillus megaterium revealed two dna regions with protein-binding capacity: +237/+318 and +319/+425 considering 'o' as the start of cytochrome p450bm-3 translation. dnasei footprint analysis of the fragment +319/+425 with the total cell extract sh ... | 1996 | 8996092 |
| survival of bacillus megaterium strains in water. | cultures of bacillus megaterium strains, producers or not of poly-beta-hydroxy-butyrate (phb+ and phb-) were submitted to several shift-downs: nutritional (one hundred fold dilution in saline water s or artificial fresh water ada) or nutritional and osmotic (one hundred fold dilution in water or w). in all conditions tested, the wild type strain survived, duplicated five times and sporulated. however, the phb- mutant strain showed a drastic loss of viability in water (< 0.1%) not observed when t ... | 1996 | 9017852 |
| monoclonal antibodies for use in detection of bacillus and clostridium spores. | five monoclonal antibodies against bacterial spores of bacillus cereus t and clostridium sporogenes pa3679 were developed. two antibodies (b48 and b183) were selected for their reactivity with b. cereus t spores, two (c33 and c225) were selected for their reactivity with c. sporogenes spores, and one (d89) was selected for its reactivity with both b. cereus and c sporogenes spores. the isotypes of the antibodies were determined to be immunoglobulin g2a (igg2a) (b48), igg1 (b183), and igm (c33, c ... | 1997 | 9023926 |
| [poly-(3-hydroxybutyrate) granules in bacillus megaterium. isolation and analysis of associated proteins]. | bacillus megaterium accumulates poly-(3-hydroxybutyrate) (phb) as a reserve material in intracellular granules. in this work we described a method for the preparation of phb granules from b. megaterium pv447 and the analysis by polyacrylamide gel electrophoresis of the associated proteins. by comparison with another species a function is proposed for some of these proteins. | 1996 | 9026821 |
| a single mutation in cytochrome p450 bm3 changes substrate orientation in a catalytic intermediate and the regiospecificity of hydroxylation. | phenylalanine 87 of bacillus megaterium cytochrome p450 bm3, a residue close to the heme in the substrate binding pocket, has been replaced by alanine by site-directed mutagenesis. the substitution had no effect on the rate of hydroxylation of laurate and increased the affinity for laurate of both the intact enzyme and its heme domain by 2.6-6-fold in the ferric state. nmr paramagnetic relaxation measurements showed that in the initial ferric enzyme-substrate complex, where the substrate binds r ... | 1997 | 9048540 |
| cbix: a novel metal-binding protein involved in sirohaem biosynthesis in bacillus megaterium. | | 1997 | 9056975 |
| regulation of expression, genetic organization and substrate specificity of xylose uptake in bacillus megaterium. | xylose uptake in bacillus megaterium depends on expression of a putative h+/xylose symporter encoded by xylt, the last gene in the xyl operon. insertional inactivation of xylt leads to an apparent uptake deficiency determined with whole cells and severely slower growth on xylose as sole carbon source. expression of xylt is xylose inducible and subject to carbon catabolite repression mediated by ccpa and cre. northern analysis of the xyl mrna reveals that a potential stem-loop structure located i ... | 1997 | 9076741 |
| cooperative and non-cooperative dna binding modes of catabolite control protein ccpa from bacillus megaterium result from sensing two different signals. | carbon catabolite repression (ccr) of several operons in bacillus subtilis and bacillus megaterium is mediated by the cis-acting cre sequence and trans-acting catabolite control protein (ccpa). we describe purification of ccpa from b. megaterium and its interaction with regulatory sequences from the xyl operon. specific interaction of ccpa with cre as scored by dnase i footprints at concentrations similar to the in vivo situation requires the presence of effectors. we have found two molecular ef ... | 1997 | 9102460 |
| adsorption of bacillus subtilis endo-beta-1,4-glucanase to cellulosic materials. | the intact and the truncated endo-beta-1,4-glucanase expressed in bacillus megaterium by a b. subtilis gene were purified and its adsorption characteristics to cellulosic materials were investigated. the intact enzyme was purified by affinity towards insoluble cellulose and mono-q chromatography. the truncated enzyme was purified by gel filtration and chromatofocusing. the molecular activities of these enzymes towards the soluble substrates were identical. optimum ph and temperature of these two ... | 1997 | 9111928 |
| steady-state and picosecond-time-resolved fluorescence studies on the recombinant heme domain of bacillus megaterium cytochrome p-450. | the conformational changes associated with the interaction of sodium laurate with the recombinant heme domain for cytochrome p-450bm3 have been investigated by steady-state and picosecond-time-resolved fluorescence spectroscopy. the steady-state quenching experiments show that while all the five tryptophan residues are accessible to acrylamide in the free enzyme as well as the enzyme x substrate complex, the number of tryptophan residues accessible to ionic quenchers decreases on interaction of ... | 1997 | 9119001 |
| transbilayer movement of fluorescent phospholipids in bacillus megaterium membrane vesicles. | we investigated the transbilayer movement or flip-flop of phospholipids in vesicles derived from the cytoplasmic membrane of bacillus megaterium. since common assay techniques were found to be inapplicable to the bacillus system, we exploited and elaborated a newly described method in which fluorescent phospholipids (1-myristoyl-2-c6-nbd phospholipids) are used as tracers to monitor flip-flop. these lipids were introduced into bacillus vesicles from synthetic donor vesicles containing a fluoresc ... | 1997 | 9125519 |
| barbiturate-induced expression of neuronal nitric oxide synthase in the rat cerebellum. | neuronal nitric oxide synthase (nnos) is known to share some structural and functional similarities with the cytochrome p450 mixed function oxidase system. unlike p450, it does not require a second enzyme, reductase, to transfer electrons. this characteristic is similar to p450(bm-3) of bacillus megaterium. p450(bm-3) and certain mammalian subfamilies of p450, such as p4502b, are known to be induced by phenobarbital (pb), and these p450s share a consensus sequence called the barbie box. because ... | 1997 | 9134969 |
| reconstitution of the fatty acid hydroxylase activity of cytochrome p450bm-3 utilizing its functional domains. | cytochrome p450bm-3, a catalytically self-sufficient fatty acid monooxygenase from bacillus megaterium, is a multidomain protein containing heme, fad, and fmn. previous attempts to reconstitute the fatty acid monooxygenase activity of intact p450bm-3 utilizing equimolar concentrations of the separate heme (bmp) and reductase (bmr) domains, have been unsuccessful because two-electron reduced fmn, which rapidly accumulates, is incapable of electron transfer to the heme iron. the present study of t ... | 1997 | 9143326 |
| the stereospecificity of hydrogen transfer to nad(p)+ catalyzed by lactol dehydrogenases. | the stereochemistry of hydrogen transfer to nad(p)+ has been determined for five lactol dehydrogenases. it was found that d-glucose dehydrogenases from bacillus megaterium and cryptococcus uniguttulatus and l-rhamnose dehydrogenase from aureobasidium pullulans are pro-s (b) specific, while d-glucose dehydrogenase from thermoplasma acidophilum and d-xylose dehydrogenase from procine liver are pro-r (a) specific. the latter two enzymes are the first examples of a-specific dehydrogenases oxidizing ... | 1997 | 9168914 |
| iron and salicylate induction of cytochrome p450bm-1 in bacillus megaterium. | the effects of iron and salicylate on the expression of cytochrome p450s in bacillus megaterium were investigated in this report. immunoblot analysis showed that the addition of 4 mm ferric iron or 10 mm salicylate to the culture medium resulted in a significant increase in the p450bm-1 level, while the same condition had little effect on p450bm-3 expression. substantial induction of chloramphenicol acetyltransferase (cat) activity by iron and salicylate in b. megaterium cells bearing a p450bm-1 ... | 1997 | 9175556 |
| polarized light scattering for rapid observation of bacterial size changes. | the effect of changing growth conditions on the diameter of rod-shaped bacteria was studied in vivo with the use of polarized light scattering. the value of a ratio of scattering matrix elements was measured as a function of scattering angle at various times after nutritional "upshift" for two strains of escherichia coli cells. the peak locations of the scattering function were calibrated against the diameter for rod-shaped bacteria. the peaks moved toward smaller angles as a function of time af ... | 1997 | 9199812 |
| functional interactions in cytochrome p450bm3: flavin semiquinone intermediates, role of nadp(h), and mechanism of electron transfer by the flavoprotein domain. | cytochrome p450bm3 is a self-sufficient soluble fatty acid hydroxylase from bacillus megaterium utilizing tightly bound fad and fmn cofactors to transfer reducing equivalents from nadph to the heme active site. active-inactive transitions of cytochrome p450bm3 were exploited to identify catalytic intermediates of the enzyme. shortly upon reduction by nadph, a two-electron reduced active p450bm3 is formed with two flavin semiquinones, anionic and neutral, present simultaneously. p450bm3 inactivat ... | 1997 | 9204888 |
| plasmids for efficient single-copy gene cloning into gdh2 and trpc of bacillus megaterium dsm319 and qm b1551. | we constructed integrative plasmids to place xyla-lacz indicator gene fusions into two different loci of the bacillus megaterium chromosome, gdh2 and trpc, in lac mutants of strains dsm 319 and qm b1551, which differ markedly. single-crossover integration was achieved in all cases while double crossovers occurred only in gdh2 of dsm 319 and qm b1551 and in trpc of qm b1551. neither of the loci affected regulation of the xyla-lacz fusions. these results confirm the suitability of the two gene loc ... | 1997 | 9210344 |
| the lantibiotic mersacidin inhibits peptidoglycan biosynthesis at the level of transglycosylation. | the lantibiotic mersacidin has been previously reported to interfere with bacterial peptidoglycan biosynthesis, [brötz, h., bierbaum, g., markus, a., molitor, e. & sahl, h.-g. (1995) antimicrob. agents chemother. 39, 714-719]. here, we focus on the target reaction and describe a mersacidin-induced accumulation of udp-n-acetylmuramoyl-pentapeptide, indicating that inhibition of peptidoglycan synthesis occurs after the formation of cytoplasmic precursors. in vitro studies involving a wall-membrane ... | 1997 | 9210483 |
| isolation of bacillus megaterium mutants that produce high levels of heterologous protein, and their use to construct a highly mosquitocidal strain. | a xylose-regulated plasmid expression system for producing high levels of recombinant proteins in bacillus megaterium has recently been described [appl microbiol biotechnol 35:594, 1991]. using an antibiotic resistance protein as the expressed protein, we have been able to select mutant plasmids that produce increased levels of heterologous protein. the mutant plasmids show increased segregational stability and have lost the ability to be transformed into escherichia coli. the same selection pro ... | 1997 | 9216879 |
| sequence and expression of an eisenia-fetida-derived cdna clone that encodes the 40-kda fetidin antibacterial protein. | fetidins are 40-kda and 45-kda hemolytic and antibacterial glycoproteins present in the coelomic fluid of the earthworm eisenia fetida andrei. by screening a cdna library with a polyclonal antifetidin serum, we have cloned a cdna that encoded the 40-kda fetidin. the clone contains an insert of 1.44 kb encoding a protein of 34 kda, which corresponds to the size of deglycosylated fetidins. the recombinant protein inhibits bacillus megaterium growth. restriction fragment polymorphisms were observed ... | 1997 | 9219536 |
| analysis of ccpa mutations defective in carbon catabolite repression in bacillus megaterium. | five mutations in ccpa of bacillus megaterium with impaired functions were analysed for carbon catabolite repression. the phenotypes support the hypothesis that ccpa assumes a purr/laci fold. the completely inactive mutants ccpa119ge and ccpa326am cause alterations which are incompatible with that fold. a mutation with reduced activity, ccpa81ge, affects a site that would be partially surface exposed and may interfere with structure formation or cofactor binding. a mutation in the putative hinge ... | 1997 | 9252590 |
| active site analysis of p450 enzymes: comparative magnetic circular dichroism spectroscopy. | recent structural studies indicate that the substrate- and o2-binding distal pocket of the p450 enzymes are not identical. thus, p450terp (cyp108) from the alpha-terpineol-metabolizing pseudomonad differs from p450cam (cyp-101) (c. a. hasemann et al., j. mol. biol. 236, 1169, 1994). in contrast, the distal pockets of p450terp and p450bmp (cyp102 heme domain; bacillus megaterium) are more closely similar, including novel hydrogen-bonding interactions between the distal h2o ligand and the i helix ... | 1997 | 9281314 |
| molecular dynamics study of time-correlated protein domain motions and molecular flexibility: cytochrome p450bm-3. | time-correlated atomic motions were used to characterize protein domain boundaries from atomic coordinates generated by molecular dynamics simulations. a novel application of the dynamical cross-correlation matrix (dccm) analysis tool was used to help identify putative protein domains. in implementing this new approach, several dccm maps were calculated, each using a different coordinate reference frame from which protein domain boundaries and protein domain residue constituents could be identif ... | 1997 | 9284282 |
| [antibodies against bacillus megaterium h glycoprotein in patients with oncological and nononcological pathology of gastro-intestinal tract]. | | 1994 | 9296708 |
| production and characterization of xylanase from a beta-amylolytic strain of bacillus megaterium. | bacillus megaterium b6 atcc 51946, a potent beta-amylase producing strain produced extracellular xylanase (ec 3.2.1.8) when cultivated in the presence of xylan as sole carbon source. the strain showed maximum xylanolytic activity after 12 h growth, and was capable of fermenting various agricultural wastes, of which under-utilized jute stalk proved to be best for production of xylanase. the ultrafiltered xylanase showed temperature and ph optima at 85 degrees c and 7.5, respectively. the enzyme w ... | 1997 | 9301068 |
| antioxidant-mediated attenuation of the induction of cytochrome p450bm-3(cyp102) by ibuprofen in bacillus megaterium atcc 14581. | bacillus megaterium contains a soluble cytochrome p450 termed bm-3, which is highly inducible by barbiturates, peroxisome proliferators, and nonsteroidal antiinflammatory drugs. in rats and mice, the chronic administration of peroxisome proliferators induces a sustained oxidative stress in hepatic tissue and may be responsible for the nongenotoxic carcinogenesis observed with prolonged treatment. here it is shown that ibuprofen induces a variety of enzymes associated with the oxidative stress re ... | 1997 | 9313770 |
| catabolite repression in lactobacillus casei atcc 393 is mediated by ccpa. | the chromosomal ccpa gene from lactobacillus casei atcc 393 has been cloned and sequenced. it encodes the ccpa protein, a central catabolite regulator belonging to the laci-galr family of bacterial repressors, and shows 54% identity with ccpa proteins from bacillus subtilis and bacillus megaterium. the l. casei ccpa gene was able to complement a b. subtilis ccpa mutant. an l. casei ccpa mutant showed increased doubling times and a relief of the catabolite repression of some enzymatic activities, ... | 1997 | 9352913 |
| fatty acid signals in bacillus megaterium are attenuated by cytochrome p-450-mediated hydroxylation. | in previous publications [english, hughes and wolf (1994) j. biol. chem. 269, 26836-26841; english, hughes and wolf (1996) biochem. j. 316, 279-283], we have demonstrated that peroxisome proliferators and non-steroidal anti-inflammatory drugs are inducers of the cytochrome p-450bm-3 gene in bacillus megaterium atcc14581. their mechanism of action involves binding to and subsequent displacement of the transcriptional repressor, bm3r1, from its operator site, which results in the activation of cyt ... | 1997 | 9359402 |
| engineering the substrate specificity of bacillus megaterium cytochrome p-450 bm3: hydroxylation of alkyl trimethylammonium compounds. | oligonucleotide-directed mutagenesis has been used to replace arginine-47 with glutamate in cytochrome p-450 bm3 from bacillus megaterium and in its haem domain. the mutant has been characterized by sequencing, mass spectrometry, steady-state kinetics and by optical and nmr measurements of substrate binding. the mutant retains significant catalytic activity towards c12-c16 fatty acids, catalysing hydroxylation in the same (omega-1, omega-2, omega-3) positions with kcat/km values a factor of 14-2 ... | 1997 | 9359427 |
| redox control of the catalytic cycle of flavocytochrome p-450 bm3. | flavocytochrome p-450 bm3 from bacillus megaterium is a 119 kda polypeptide whose heme and diflavin domains are fused to produce a catalytically self-sufficient fatty acid monooxygenase. redox potentiometry studies have been performed with intact flavocytochrome p-450 bm3 and with its component heme, diflavin, fad, and fmn domains. results indicate that electron flow occurs from the nadph donor through fad, then fmn and on to the heme center where fatty acid substrate is bound and monooxygenatio ... | 1997 | 9374858 |
| replacement of isoleucine-47 by threonine in the hpr protein of streptococcus salivarius abrogates the preferential metabolism of glucose and fructose over lactose and melibiose but does not prevent the phosphorylation of hpr on serine-46. | phosphorylation of hpr on a serine residue at position 46 (ser-46) by an atp-dependent protein kinase has been reported in several gram-positive bacteria, and the resulting intermediate, hpr(ser-p), has been shown to mediate inducer exclusion in lactococci and lactobacilli and catabolite repression in bacillus subtilis and bacillus megaterium. we report here the phenotypic properties of an isogenic spontaneous mutant (g22.4) of streptococcus salivarius atcc 25975, in which a missense mutation re ... | 1997 | 9379899 |
| contribution of glucose kinase to glucose repression of xylose utilization in bacillus megaterium. | the glk gene from bacillus megaterium, which encodes glucose kinase, was isolated and analyzed. disruption by a transcriptional glk-luxab fusion indicated that glk is the only glucose kinase gene in that strain but did not affect growth of that mutant on glucose. determination of luciferase activity under various growth conditions revealed constitutive transcription of glk. expression of a xyla-lacz fusion was repressed by glucose in the strain with the glk disruption about twofold less efficien ... | 1997 | 9393732 |
| thr268 in substrate binding and catalysis in p450bm-3. | members of the gene superfamily of proteins called "p450" catalyze monooxygenation reactions that require an input of two electrons and a molecule of oxygen per catalytic cycle. these proteins are widely distributed among living organisms, from bacteria to human. p450bm-3, a soluble protein isolated from bacillus megaterium, is self-sufficient, containing p450 and reductase domains on the same polypeptide. p450bm-3 catalyzes the hydroxylation of various fatty acids at omega-1, omega-2, and omega ... | 1998 | 9439582 |
| t-cell response during rhodococcus equi infection in a murine experimental model. | rhodococcus equi is a facultative intracellular bacterium that can cause pneumonia in both young horses and immunocompromised humans. in this study, we have tried to determine the t-cell populations that recognize this pathogen during murine infection, as well as the bacterial antigens recognized by these cells. when balb/c mice were hyperimmunized with a virulent r. equi strain, we did not observe preferential expansion of a particular t-cell subset in their spleens. however, when the splenic t ... | 1997 | 9443578 |
| redox characterisation of flavocytochrome p-450 bm3 from bacillus megaterium. | | 1997 | 9450056 |
| thermostable, high salt-tolerant amylase from bacillus megaterium vumb-109. | a bacterial strain, bacillus megaterium vumb-109, has been isolated and identified which produces salt-tolerant, thermostable amylase (16 u/ml culture filtrate). cultural conditions such as carbon and nitrogen sources, metal ions, temperature and ph have been optimized for enzyme production. the partially purified enzyme was active over a wide range of ph (4.5-10) and exhibited maximum activity at 95 degrees c, retaining 90% original activity at 100 degrees c. partially purified enzyme was stabl ... | 1997 | 9468732 |
| escherichia coli transformant expressing the glucose dehydrogenase gene from bacillus megaterium as a cofactor regenerator in a chiral alcohol production system. | escherichia coli jm109 (pgda2) overexpressing the glucose dehydrogenase (gdh) gene from bacillus megaterium iwg3 was examined for use as a cofactor regenerator. in the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate by e. coli jm109 (pkar) which is an aldehyde reductase-overproducing transformant, e. coli jm109 (pgda2) can act as an nadph regenerator with nadp+ and glucose, similarly to commercially available gdh. | 1998 | 9501530 |
| beta-amylase production by some bacillus cereus, bacillus megaterium and bacillus polymyxa [correction of polymaxa] strains. | the production of extracellular beta-amylase by some bacillus cereus, bacillus megaterium and bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 u/ml of the culture fluid, respectively (u, 1 mumol of maltose equivalent per min at 30 degrees c). several cultivation media were used for beta-amylase production. bacillus cereus and some strains of bacillus megaterium gave good yields of beta-amylase only in medium with the addition of ... | 1997 | 9516983 |
| evidence against the bm1p1 protein as a positive transcription factor for barbiturate-mediated induction of cytochrome p450bm-1 in bacillus megaterium. | the bm1p1 protein was previously proposed to act as a positive transcription factor involved in barbiturate-mediated induction of cytochrome p450bm-1 in bacillus megaterium. we now report that the bm1p1 gene encodes a protein of 217 amino acids, rather than the 98 amino acids as reported previously. in vitro gel shift assays indicate that the bm1p1 protein did not interact with probes comprising the regulatory regions of the p450bm-1 gene. moreover, disruption of the bm1p1 gene did not markedly ... | 1998 | 9525898 |
| induction of cyp102 (cytochrome p450bm-3) in bacillus megaterium by 17 beta-estradiol and 4-sec-butylphenol. | cyp102 (cytochrome p450bm-3) is induced in bacillus megaterium by barbiturates, peroxisome proliferators, and nonsteroidal anti-inflammatory drugs. we now describe the induction of cyp102 in b. megaterium by 17 beta-estradiol and by 4-sec-butylphenol. these estrogens interact with the repressor protein bm3r1, causing it to dissociate with the operator of the cyp102 gene and allowing transcription to occur. we have developed a stable transfection of a construct into b. megaterium of a truncated c ... | 1998 | 9535758 |
| in vivo roles of bm3r1 repressor in the barbiturate-mediated induction of the cytochrome p450 genes (p450(bm-3) and p450(bm-)1) of bacillus megaterium. | we previously showed [q. liang, a.j. fulco, j. biol. chem., 270 (1995) 18606-18614) that the binding of bm3r1 repressor to barbie box elements and operator sites in the 5'-flanking regions of the p450bm-3 and p450bm-1 (cyp102 and cyp106) genes in bacillus megaterium was a critical factor in their regulation at the level of transcription. we now describe experiments that delineate specific roles for bm3r1 in the barbiturate-mediated induction of these genes. we directly demonstrate the interactio ... | 1998 | 9565684 |
| gas vesicle genes identified in bacillus megaterium and functional expression in escherichia coli. | gas vesicles are intracellular, protein-coated, and hollow organelles found in cyanobacteria and halophilic archaea. they are permeable to ambient gases by diffusion and provide buoyancy, enabling cells to move upwards in liquid to access oxygen and/or light. in halobacteria, gas vesicle production is encoded in a 9-kb cluster of 14 genes (4 of known function). in cyanobacteria, the number of genes involved has not been determined. we now report the cloning and sequence analysis of an 8,142-bp c ... | 1998 | 9573198 |
| identification of protein-protein contacts between alpha/beta-type small, acid-soluble spore proteins of bacillus species bound to dna. | small, acid-soluble spore proteins (sasp) of the alpha/beta-type from several bacillus species were cross-linked into homodimers, heterodimers and homooligomers with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (edc) in the presence of linear plasmid dna. significant protein cross-linking was not detected in the absence of dna. in all four alpha/beta-type sasp examined, the amino donor in the edc induced amide cross-links was the alpha-amino group of the protein. however, the carboxylate contai ... | 1998 | 9651315 |
| pyrrole-2-carboxylate decarboxylase from bacillus megaterium pyr2910, an organic-acid-requiring enzyme. | inducible pyrrole-2-carboxylate decarboxylase, which catalyzes the decarboxylation of pyrrole-2-carboxylate to pyrrole and co2 in stoichiometric amounts, was purified from bacillus megaterium pyr2910. the purity of the enzyme was shown by sds/page and gel-permeation hlpc. the enzyme has a molecular mass of approximately 98 kda and consists of two identical subunits. it is highly specific for pyrrole-2-carboxylate, and also catalyzes the reverse reaction, the carboxylation of pyrrole. a unique fe ... | 1998 | 9654100 |
| cobalamin (vitamin b12) biosynthesis--cloning, expression and crystallisation of the bacillus megaterium s-adenosyl-l-methionine-dependent cobalt-precorrin-4 transmethylase cbif. | the bacillus megaterium cbif, encoding the cobalt-precorrin-4 s-adenosyl-l-methionine-dependent transmethylase of the anaerobic cobalamin biosynthetic pathway, has been cloned and overexpressed as a his-tagged recombinant protein in escherichia coli. the protein was purified to homogeneity by a combination of metal chelate chromatography and high-resolution anion-exchange chromatography. the protein migrated with a subunit mass of 31 kda by sds/page and with a molecular mass of 62 kda by analyti ... | 1998 | 9660189 |
| the repressor protein, bm3r1, mediates an adaptive response to toxic fatty acids in bacillus megaterium. | bm3r1 is a helix-turn-helix transcriptional repressor from bacillus megaterium whose binding to dna is inhibited by fatty acids and a wide range of compounds that modulate lipid metabolism. the inactivation of bm3r1/dna binding activity results in the activation of transcription of the operon encoding a fatty acid hydroxylase, cytochrome p450 102. the metabolic role of this operon is unknown. it is possible that it is involved in the synthesis of modified fatty acids as part of normal cellular m ... | 1998 | 9660768 |
| the bacillus spoiiga protein is targeted to sites of spore septum formation in a spoiie-independent manner. | the process of bacterial cell division involves the assembly of a complex of proteins at the site of septation that probably provides both the structural and the cytokinetic functions required for elaboration and closure of the septal annulus. during sporulation in bacillus subtilis, this complex of proteins is modified by the inclusion of a sporulation-specific protein, spoiie, which plays a direct role in gene regulation and also has a genetically separable role in determining the gross struct ... | 1998 | 9663680 |
| the x-ray structure of a cobalamin biosynthetic enzyme, cobalt-precorrin-4 methyltransferase. | biosynthesis of the corrin ring of vitamin b12 requires the action of six s-adenosyl-l-methionine (adomet) dependent transmethylases, closely related in sequence. the first x-ray structure of one of these, cobalt-precorrin-4 transmethylase, cbif, from bacillus megaterium has been determined to a resolution of 2.4 a. cbif contains two alphabeta domains forming a trough in which s-adenosyl-l-homocysteine (adohcy) binds. the location of adohcy and a number of conserved residues, helps define the pr ... | 1998 | 9665173 |
| trans activation of the escherichia coli ato structural genes by a regulatory protein from bacillus megaterium: potential use in polyhydroxyalkanoate production. | a bacillus megaterium genomic fragment, which encoded an activator homologous to sigma 54 regulators and which was capable of activating escherichia coli ato genes in trans, was detected in a gene library of b. megaterium screened for beta-ketothiolase activity. the fragment presented only one complete open reading frame (orf1), which encoded a protein of 398 amino acids. the recombinant plasmid complemented mutations in the escherichia coli atoc regulatory gene. the constitutive expression of t ... | 1998 | 9684307 |
| cloning, expression, and catabolite repression of a gene encoding beta-galactosidase of bacillus megaterium atcc 14581. | a gene encoding beta-galactosidase, designated mbga, was isolated from bacillus megaterium atcc 14581. chromosomal beta-galactosidase production could be dramatically induced by lactose but not by isopropyl-beta-d-thiogalactopyranoside (iptg) and was subject to catabolite repression by glucose. disruption of mbga in the b. megaterium chromosome resulted in loss of lactose-inducible beta-galactosidase production. a 27-bp inverted repeat was found to overlap the mbga promoter sequence. two partial ... | 1998 | 9721318 |
| microbial hydroxylation of acetylaminosteroids. | an investigation of the microbial biotransformation of a range of 3 beta-, 17 beta-, and 20-acetylamino c18 to c21 steroids by microorganisms known to hydroxylate conventional steroids has been undertaken, using aspergillus ochraceus, bacillus megaterium, curvularia lunata, and rhizoputus arrhizus. a. ochraceus and b. megaterium gave products of 11 alpha- and 15 beta-hydroxylation, respectively. in the case of c. lunata, the products were predominantly those of this organism's normal c-11 beta- ... | 1998 | 9727096 |