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new thermosensitive plasmid for gram-positive bacteria.we isolated a replication-thermosensitive mutant of the broad-host-range replicon pwv01. the mutant pve6002 is fully thermosensitive above 35 degrees c in both gram-negative and gram-positive bacteria. four clustered mutations were identified in the gene encoding the replication protein of pve6002. the thermosensitive derivative of the related plasmid pe194 carries a mutation in the analogous region but not in the same position. derivatives of the thermosensitive plasmid convenient for cloning p ...19921324906
effect of changes in urine ph on plasma pharmacokinetic variables of ampicillin sodium in horses.the effect of urine ph on plasma disposition of ampicillin sodium was evaluated. a single dose of 10 mg/kg of body weight was administered iv to thoroughbreds with alkaline (ph greater than 8.0) or acidic (ph less than 4.5) urine. urine alkalinity was achieved and maintained by oral administration of up to 400 mg of sodium bicarbonate/kg/d, and acidity was achieved and maintained by oral administration of up to 400 mg of ammonium chloride/kg/d. ampicillin sodium was measured in the plasma of hor ...19921326242
optical and resonance raman spectroscopy of the heme groups of the quinol-oxidizing cytochrome aa3 of bacillus subtilis.the cytochrome aa3-type terminal quinol oxidase of bacillus subtilis catalyzes the four-electron reduction of dioxygen to water. it resembles the aa3-type cytochrome-c oxidase in using heme a as its active-site chromophores but lacks the cua center and the cytochrome-c oxidizing activity of the mitochondrial enzyme. we have used optical and resonance raman spectroscopies to study the b. subtilis oxidase in detail. the alpha-band absorption maximum of the reduced minus oxidized enzyme is shifted ...19921327130
identification of tms-26 as an allele of the gcad gene, which encodes n-acetylglucosamine 1-phosphate uridyltransferase in bacillus subtilis.the temperature-sensitive bacillus subtilis tms-26 mutant strain was characterized biochemically and shown to be defective in n-acetylglucosamine 1-phosphate uridyltransferase activity. at the permissive temperature (34 degrees c), the mutant strain contained about 15% of the wild-type activity of this enzyme, whereas at the nonpermissive temperature (48 degrees c), the mutant enzyme was barely detectable. furthermore, the n-acetylglucosamine 1-phosphate uridyltransferase activity of the tms-26 ...19921328164
determination of sparfloxacin in serum and urine by high-performance liquid chromatography.a specific and sensitive analytical method for the determination of sparfloxacin in serum and urine is described. serum proteins are removed by precipitation with acetonitrile after the addition of ofloxacin as an internal standard. the supernatant solvent is evaporated in a vacuum concentrator and the dry residue is redissolved in the mobile phase. separation is performed on a cation-exchange column (nucleosil 100 5sa, 125 x 4.0 mm i.d., 5 microns particle size) protected by a guard column (per ...19921331140
metal-based formulations with high microbicidal activity.substances were evaluated for their relative potencies in inactivating junin virus, escherichia coli, and spores of bacillus subtilis. under the conditions of our test, glutaraldehyde was the most efficient agent among all substances currently recommended for disinfecting and sterilizing medical devices. either copper or iron ions by themselves were able to inactivate virus with an efficiency similar to that of substances currently used for disinfection and sterilization. the microbicidal effect ...19921332611
dna repair and the evolution of transformation in haemophilus influenzae.under certain environmental conditions, naturally transforming bacteria are induced to pick up dna released into the environment by other cells of the same or closely related species and, by homologous recombination, integrate that dna into their chromosome. the selective pressures responsible for the evolution and maintenance of this form of genetic outcrossing, or sex, in bacteria are not known. a prominent hypothesis is that transformation, and sex in general, evolved as a means of obtaining ...19921334020
cloning and sequencing of is1086, an alcaligenes eutrophus insertion element related to is30 and is4351.a new insertion sequence (is), designated is1086, was isolated from alcaligenes eutrophus ch34 by being trapped in plasmid pjv240, which contains the bacillus subtilis sacb and sacr genes. the 1,106-bp is1086 element contains partially matched (22 of 28 bp) terminal-inverted repeats and a long open reading frame. hybridization data suggest the presence of one copy of is1086 in the strain ch34 heavy-metal resistance plasmid pmol28 and at least two copies in its chromosome. analysis of the is1086 ...19921334071
tetracycline enhances tn916-mediated conjugal transfer.pregrowth of the donor on medium containing tetracycline increased conjugative transposition of tn916 and the transposon-dependent mobilization of pc194 19- to 119-fold in matings between bacillus subtilis and bacillus thuringiensis subsp. israelensis. tn916 and pc194 transferred independently under these conditions. when enterococcus faecalis was the donor and b. thuringiensis subsp. israelensis the recipient, pregrowth in tetracycline increased the conjugative transposition frequency by approx ...19921334267
the membrane-induced proton motive force influences the metal binding ability of bacillus subtilis cell walls.bacillus subtilis 168 is a gram-positive bacterium whose cell wall contains the highly electronegative polymers peptidoglycan (chemotype a1 gamma) and glycerol-based teichoic acid to produce a surface with a net negative charge with high metal binding capacity. during metabolism, a membrane-induced proton motive force continuously pumps protons into the wall fabric. as a result, a competition between protons and metal ions for anionic wall sites occurs, and less metal is bound in living cells th ...19921335717
utilisation of glycerol and glycerol 3-phosphate is differently affected by the phosphotransferase system in bacillus subtilis.glycerol and glycerol 3-phosphate uptake in bacillus subtilis does not involve the phosphotransferase system. in spite of this, b. subtilis mutants defective in the general components of the phosphotransferase system, enzymei or hpr, are unable to grow with glycerol as sole carbon and energy source. here we show that a hpr mutant can grow on glycerol 3-phosphate and that glycerol 3-phosphate, but not glycerol, can induce glpd encoding glycerol-3-phosphate dehydrogenase. induction of glpd also re ...19921335945
molecular biology of bacillus subtilis cytochromes.bacillus subtilis cells must have cytochromes for growth and can synthesize cytochromes of a-, b-, c-, d-, and o-types. after a long lag, our knowledge of the structure, genetics and specific role for these cytochromes is now growing exponentially as the result of recent research. this progress is reviewed here and includes, for example, the discovery of two different cytochrome a systems and genes required for their biogenesis.19921335950
the lux genes in photobacterium leiognathi are closely linked with genes corresponding in sequence to riboflavin synthesis genes.three open reading frames (orfs) have been found in the region downstream of the luxg gene in the photobacterium leiognathi lux operon. these genes (orf i, ii, and iii) are not only closely linked to the lux operon and transcribed in the same direction but also show the same organization and code for proteins homologous in sequence to the gene products of ribb, riba, and ribh of bacillus subtilis, respectively. the photobacterium leiognathi gene (orf ii) corresponding to riba was expressed in es ...19921339274
nuclear-encoded chloroplast ribosomal protein l27 of nicotiana tabacum: cdna sequence and analysis of mrna and genes.a tobacco (nicotiana tabacum cv. petite havana) leaf cdna library was constructed in the expression vector lambda gt11. immunological and nucleic acid hybridization screening yielded several cdnas encoding an m(r) 19,641 precursor to an m(r) 14,420 mature protein which is homologous to escherichia coli ribosomal protein l27. one cdna (l27-1; 882 nucleotides long) contains 104 bp of 5'-noncoding sequence, 51 codons for a transit peptide, 128 codons for the predicted mature l27 polypeptide, and 24 ...19921339289
characterization and properties of a novel plasmid vector for bacillus thuringiensis displaying compatibility with host plasmids.a novel plasmid vector, composed of a 1.7-kb bacillus thuringiensis (b.t.) replicon, a multiple cloning site, and an erythromycin-resistance marker gene from bacillus subtilis, was constructed for use in b.t. unlike other vectors which have been reported to be acceptable for b.t., this new b.t. vector was stably maintained in the absence of er and did not displace host plasmids, some of which carry crystal protein-encoding genes (cry genes). the compatibility of this b.t. vector with native plas ...19921339372
cloning, sequencing, and molecular analysis of the dnak locus from bacillus subtilis.by using an internal part of the dnak gene from bacillus megaterium as a probe, a 5.2-kb hindiii fragment of chromosomal dna of bacillus subtilis was cloned. downstream sequences were isolated by in vivo chromosome walking. sequencing of 5,085 bp revealed four open reading frames in the order orf39-grpe-dnak-dnaj. orf39 encodes a 39-kda polypeptide of unknown biological function with no noticeable homology to any other protein within the data bases. alignment of the grpe protein with those of th ...19921339421
characterization of the streptomyces clavuligerus argc gene encoding n-acetylglutamyl-phosphate reductase: expression in streptomyces lividans and effect on clavulanic acid production.the argc gene of streptomyces clavuligerus encoding n-acetylglutamyl-phosphate reductase (agpr) has been cloned by complementation of argc mutants streptomyces lividans 1674 and escherichia coli xc33. the gene is contained in an open reading frame of 1,023 nucleotides which encodes a protein of 340 amino acids with a deduced molecular mass of 35,224 da. the argc gene is linked to arge, as shown by complementation of arge mutants of e. coli. expression of argc from cloned dna fragments carrying t ...19921339424
[cloning and gene expression of bacillus cereus neutral proteinase in bacillus subtilis cells].the neutral proteinase gene of bacillus cereus was cloned. its restriction map and the direction of transcription was determined. it was shown that the neutral proteinase gene could be expressed in bacillus cells. the thermostability of the product coded by the neutral proteinase gene and its natural analogue was explored. the obtained data indicate that the neutral proteinase of bacillus cereus is closely related to the enzyme of bacillus amyloliquefaciens by these parameters. it was found that ...19921339956
[biological products and laboratory reagents dehydrated by a rapid direct vacuum desiccation method. v. the preservation of the biological product enterosubtil i.c. by direct desiccation]. 19921340259
"bacillus subtilis" infection in a patient submitted to a bone marrow transplantation. 19921340996
[ethylene oxide sterilization. i. influence of the sporulation medium on the resistance of spores of bacillus subtilis var. niger].some elements influencing the resistance of spores used in ethylene oxide sterilization process control are standardized. spores of bacillus subtilis var. niger were produced in chemically defined liquid and solid sporulation media to a total of 12 lots; after standardization of the number of spores, they were challenged by sub-lethal cycles, followed by a lethality study. according to the statistical model applied, there were no differences between the resistance of spores produced in chemicall ...19921342528
[ethylene oxide sterilization. ii. influence of test devices on the performance of biological monitors and its evaluation].in view of the importance of the assurance of the sterility of medical devices, with a large incidence of tubular forms, a study of biological monitors was undertaken, using paper as a carrier, the dimension of the test specimens being the variable considered. after the sterilization process in an industrial cycle, followed by the recovery of the surviving spores through inoculation of the carriers into thioglycolate broth, soybean-casein broth and this last with the addition of bromothymol blue ...19921342529
sexuality in a natural population of bacteria--bacillus subtilis challenges the clonal paradigm.reproduction by binary fission necessarily establishes a clonal genotypic structure in bacterial populations unless a high rate of genetic recombination opposes it. several genetic properties were examined for a wild population of bacillus subtilis in the sonoran desert of arizona to assess the extent of recombination in a natural population. these properties included allozyme variation revealed by multilocus enzyme electrophoresis, phage and antibiotic resistance, and restriction fragment lengt ...19921344989
[preparation and use of fish fillet infusion as a basic medium for culturing bacteria].the authors present the first results on the utilization of fish infusion (ifp) as a basic medium for the cultivation of bacteria. the infusion was obtained from a common marine fish, corvina (micropogonias furnieri) according to the technique used in the preparation of beef infusion broth. streptococcus pyogenes, s. pneumoniae, neisseria meningitidis, campylobacter jejuni, escherichia coli, klebsiella pneumoniae, serratia marcescens, pseudomonas aeruginosa, staphylococcus aureus and bacillus su ...19921345299
analysis of the autolysins of bacillus subtilis 168 during vegetative growth and differentiation by using renaturing polyacrylamide gel electrophoresis.the autolysins of bacillus subtilis 168 were analyzed by sodium dodecyl sulfate (sds)-polyacrylamide gel electrophoresis with substrate-containing gels. four bands of vegetative autolytic activity of 90, 50, 34, and 30 kda (bands a1 to a4) were detected in sds and licl extracts and in native cell walls by using b. subtilis 168 vegetative cell walls as the substrate incorporated in the gel. the four enzyme activities showed different substrate specificities and sensitivities to various chemical t ...19921345911
interaction of the bacillus subtilis glnra repressor with operator and promoter sequences in vivo.in vivo dimethyl sulfate footprinting of the bacillus subtilis glnra regulatory region under repressing and derepressing conditions demonstrated that the glnr protein, encoded by glnr, interacts with two sites situated within and adjacent to the glnra promoter. one site, glnrao1, between positions -40 and -60 relative to the start point of transcription, is a 21-bp symmetrical element that has been identified as essential for glnra regulation (h. j. schreier, c. a. rostkowski, j. f. nomellini, a ...19921346263
the glna gene of the extremely thermophilic eubacterium thermotoga maritima: cloning, primary structure, and expression in escherichia coli.the structural gene (glna) encoding the glutamine synthetase (gs) of the extremely thermophilic eubacterium thermotoga maritima has been cloned on a 6.0 kb hindiii dna fragment. sequencing of the region containing the glna gene (1444 bp) showed an orf encoding a polypeptide (439 residues) with an estimated mass of 50,088 da, which shared significant homology with the gsi sequences of other bacteria (escherichia coli, bacillus subtilis) and archaea (pyrococcus woesei, sulfolobus solfataricus). th ...19921348781
typing of listeria monocytogenes by restriction polymorphism of the ribosomal ribonucleic acid gene region.ninety four strains of listeria monocytogenes of different serovars and phagovars as well as of varying origins were characterized by ribosomal rna gene restriction polymorphism. after digestion by ecori or hindiii, chromosomal dnas were hybridized with a cloned rdna probe from bacillus subtilis that included the 16s rrna gene. the 94 strains were divided into 14 ribovars according to the different hybridization patterns generated by cleavage with ecori. less important genomic heterogeneity coul ...19921349500
isolation and characterization of the major vegetative rna polymerase of streptomyces coelicolor a3(2); renaturation of a sigma subunit using groel.the promoter region of the agarase gene (daga) of streptomyces coelicolor a3(2) is complex; it consists of four distinct promoters with different -10 and -35 regions. we report the isolation of a form of rna polymerase that mediates transcription in vitro from the dagap4 promoter. the core components of this rna polymerase are associated with a polypeptide of c. 66 kda; holoenzyme reconstitution experiments show that the 66 kda polypeptide functions as a sigma factor that directs transcription f ...19921350315
gadolinium ion inhibits loss of metabolites induced by osmotic shock and large stretch-activated channels in bacteria.bacteria subjected to a hypotonic osmotic shock lose internal ions and also metabolites, without lysis of the cells. we show that the presence in the shock medium, at submillimolar concentrations, of the ion gadolinium, recently shown to block stretch-activated channels in xenopus oocytes [yang, x.-c. & sachs, f. (1989) science 243, 1068-1071], was sufficient to inhibit shock-induced release of metabolites such as lactose and atp in escherichia coli and atp in streptococcus faecalis. moreover, g ...19921350764
cloning and characterization of the groesl operon from bacillus subtilis.the sequence of the 10 n-terminal amino acids of a bacillus subtilis protein that cross-reacts with antibody to escherichia coli groel was used to design a set of degenerate oligonucleotide probes. these probes identified a clone which carries almost the entire groesl operon from a b. subtilis subgenomic library. by chromosomal walking, an additional fragment carrying the 3' end of groesl and its flanking sequence was isolated. sequence analysis revealed two open reading frames (orfs) in the clo ...19921350776
cloning, sequencing, mapping, and transcriptional analysis of the groesl operon from bacillus subtilis.using a gene probe of the escherichia coli groel gene, a 1.8-kb hindiii fragment of chromosomal dna of bacillus subtilis was cloned. upstream sequences were isolated as a 3-kb psti fragment. sequencing of 2,525 bp revealed two open reading frames in the order groes groel. alignment of the groes and groel proteins with those of eight other eubacteria revealed 50 to 65% and 72 to 84% sequence similarity, respectively. primer extension studies revealed one potential transcription start site precedi ...19921350777
characterization of the pcp gene encoding the pyrrolidone carboxyl peptidase of bacillus subtilis.pyrrolidone carboxyl peptidase (ec 3.4.11.8) (pcp) is an enzyme that catalyzes the removal of the n-terminal pyroglutamyl group from some peptides or proteins. its value in protein chemistry and bacterial diagnosis makes this enzyme an interesting subject of study. the present paper reports for the first time the cloning and characterization of a pyrrolidone carboxyl peptidase gene (pcp). this gene is present in a single copy in the genome of bacillus subtilis as indicated by southern blot hybri ...19921353026
evaluation of a ribosomal rna gene probe for the identification of species and subspecies within the genus staphylococcus.to evaluate a 16s rrna gene probe for the identification of staphylococcal species and subspecies, we have augmented previous studies involving 12 staphylococcal species by analysing the remaining 16 species currently classified in the genus staphylococcus. hindiii- and ecori-restricted dna of isolates from validly described species of staphylococcus was probed with radiolabelled plasmid pba2 containing 16s rdna from bacillus subtilis. the dice coefficient was used to assess similarity between t ...19921353785
bacteriophage phi 105clz induces the groel-homologue protein in bacillus subtilis.using two-dimensional polyacrylamide gel electrophoresis, the groel homologue of bacillus subtilis was shown to be induced upon infection with phi 105clz, a clear plaque mutant of the temperate bacteriophage phi 105. western blotting of one dimensional polyacrylamide gels also showed the induction of the groel homologue when cells were infected with phi 105clz.19921353952
streptomyces lividans possesses a groel-like chaperonin.streptomyces lividans grown at 45 degrees c produces a groel-like chaperonin. this protein is specifically synthesized in bacterial cell cultures upon heat shock induction. it has a similar size (62 kda) to the groel-like proteins from escherichia coli and bacillus subtilus and shows immunological cross-reaction with serum raised against groel from e. coli. the s. lividans 62-kda protein assembles into oligomers around 20s that show a morphology consistent with a barrel showing six-fold and seve ...19921354626
pulo, a component of the pullulanase secretion pathway of klebsiella oxytoca, correctly and efficiently processes gonococcal type iv prepilin in escherichia coli.the pulo protein required for extracellular secretion of pullulanase by klebsiella oxytoca is known to be highly homologous to two type iv prepilin peptidases, namely xcpa(pild) (pseudomonas aeruginosa) and tcpj (vibrio cholerae). the predicted prepilin peptidase activity of pulo was confirmed by showing that it could correctly process the product of the cloned pile.1 type iv pilin structural gene from neisseria gonorrhoeae in escherichia coli. the p. aeruginosa prepilin peptidase and another pu ...19921354833
characterization of the bacillus subtilis cwba protein which stimulates cell wall lytic amidases.the bacillus subtilis cell wall binding protein, cwba, stimulated the cell wall lytic activities of the b. subtilis and b. licheniformis autolysins (cwla and cwlm, respectively) in addition to that of the major b. subtilis autolysin (cwlb). even though the substrate for the enzyme reaction was changed from b. subtilis cell wall containing a teichoic acid to micrococcus luteus cell wall containing a teichuronic acid, the stimulatory effect of cwba on cwla activity was observed.19921355454
functional analysis of the intramolecular chaperone. mutational hot spots in the subtilisin pro-peptide and a second-site suppressor mutation within the subtilisin molecule.the n-terminal pro-peptide of 77 amino acid residues is essential for the folding of subtilisin, an alkaline serine protease from bacillus subtilis. the synthetic pro-peptide has been shown to be capable of guiding the proper folding of denatured subtilisin to enzymatically active enzyme. thus the pro-peptide serves as an intramolecular chaperone, which is removed by an autoprocessing reaction after the completion of the folding. with use of localized polymerase chain reaction random mutagenesis ...19921355566
molecular cloning and sequencing of the upstream region of the major bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoiid product.the upstream region of the n-acetylmuramoyl-l-alanine amidase gene (cwlb; a major bacillus subtilis autolysin) was cloned into escherichia coli by chromosome walking. sequencing of the region showed the presence of two open reading frames, one (designated as cwba) which starts at a uug codon and encodes a polypeptide of 705 amino acids with an m(r) of 76,725, and the other (designated as lppx), upstream of cwba, comprising 102 amino acids and having a signal sequence characteristic of a lipoprot ...19921356138
in archaebacteria, there is a doxorubicin efflux pump similar to mammalian p-glycoprotein.we selected for study an anthracycline-resistant mutant from the archaebacteria haloferax volcanii. this resistance was reversed by a ca(2+)-channel antagonist, nifedipine (ndp). this resistance and its reversal by ndp suggest p-glycoprotein (pgp) to be responsible for maintaining an anticancer drug concentration below the cytotoxic level. using rhodamine 123 (rh123) as a substrate for pgp, we then examined whether the resistance to anthracyclines in this bacteria might involve a pgp-like anthra ...19921356441
usefulness of the id32 staph system and a method based on rrna gene restriction site polymorphism analysis for species and subspecies identification of staphylococcal clinical isolates.the usefulness of the id32 staph system and a method based on rrna gene restriction site polymorphism was evaluated by the study of 42 staphylococcal clinical isolates phenotypically difficult to identify. the id32 staph micromethod and the genomic method are adapted for recognition of 27 and 31 staphylococcal taxa, respectively. the genomic method is based on a dice analysis of the hybridization patterns obtained by cutting the cellular dna either with ecori or with hindiii and by probing with ...19921357001
effects of trypanothione on the biological activity of irradiated transforming dna.held et al. (1984a,b) demonstrated previously that glutathione (gsh), a negatively charged thiol, is significantly less efficient in the hydrogen atom donation repair reaction with radicals induced by radiation in transforming dna (t-dna) than are other thiol compounds. fahey et al. (1991a,b) postulated that the charge on thiols can influence their ability to radioprotect dna. gsh, which is excluded from the vicinity of dna due to its negative charge, is less protective than neutral or positivel ...19921357053
sequencing and analysis of the bacillus subtilis lytrabc divergon: a regulatory unit encompassing the structural genes of the n-acetylmuramoyl-l-alanine amidase and its modifier.the regulatory unit of bacillus subtilis strain 168 encompassing the structural genes of the n-acetylmuramoyl-l-alanine amidase and of its modifier has been sequenced, and found to be a divergon consisting of divergently transcribed operons lytabc and lytr. proteins lyta, lytb and lytc are endowed with export signal peptides. mature lyta is a 9.4 kda, highly acidic polypeptide whose deduced amino acid sequence points to a lipoprotein. lytb and lytc, the modifier and the amidase, are highly basic ...19921357079
molecular characterization of pcp, the structural gene encoding the pyrrolidone carboxylyl peptidase from streptococcus pyogenes.this paper describes the cloning of a gene (pcp) coding for pyrrolidone carboxylyl peptidase (pyrase), an enzyme which selectively removes n-terminal pyroglutamic acid residues from polypeptides. this gene was isolated from streptococcus pyogenes by construction of a gene library with a bacteriophage lambda-derived cosmid-escherichia coli host system. nucleotide sequence determination of a 1.3 kb restriction fragment revealed a 645 bp open reading frame encoding a 215-amino-acid product of m(r) ...19921357525
effect of the weak ca(2+)-binding site of subtilisin j by site-directed mutagenesis on heat stability.the functional role of the negatively charged amino acid residue in subtilisin j from bacillus stearothermophilus has been investigated by site-directed mutagenesis. glu-195 located at the weak ca2+-binding site was replaced with gln to examine the role of glu-195 in the heat stability of subtilisin j. mutant enzyme was expressed in bacillus subtilis and was purified from the culture supernatant. when the mutant enzyme was expressed at 37 degrees c in the presence of 2mm calcium chloride, the pa ...19921358066
organization and nucleotide sequence of the glutamine synthetase (glna) gene from lactobacillus delbrueckii subsp. bulgaricus.a 3.3-kb bamhi fragment of lactobacillus delbrueckii subsp. bulgaricus dna was cloned and sequenced. it complements an escherichia coli glna deletion strain and hybridizes strongly to a dna containing the bacillus subtilis glna gene. dna sequence analysis of the l. delbrueckii subsp. bulgaricus dna showed it to contain the glna gene encoding class i glutamine synthetase, as judged by extensive homology with other prokaryotic glna genes. the sequence suggests that the enzyme encoded in this gene ...19921359838
analysis of the transcriptional activity of the hut promoter in bacillus subtilis and identification of a cis-acting regulatory region associated with catabolite repression downstream from the site of transcription.levels of transcripts initiated at a hut promoter in bacillus subtilis were analysed. the addition of histidine to the culture medium increased the level of the transcript sixfold. in the presence of histidine and glucose together, the level of the transcript was reduced to the level in the absence of induction. furthermore, addition of a mixture of 16 amino acids to cultures of induced cells and of catabolite-repressed cells decreased levels of the transcript 16-fold and 2.6-fold, respectively. ...19921360137
stress proteins and cross-protection by heat shock and salt stress in bacillus subtilis.bacillus subtilis induced a set of general stress proteins in response to a salt or heat stress. cells subjected to a mild heat stress showed a protective response which enabled them to survive otherwise lethal temperatures (e.g. 52 degrees c). in a similar way bacteria were enabled to survive toxic concentrations of nacl by pretreatment with lower salt concentrations. a mild heat shock induced a cross-protection against lethal salt stress. the pretreatment of cells with low salt, however, was l ...19921362210
purification and characterization of recombinant pyrrolidone carboxyl peptidase of bacillus subtilis.bacillus subtilis pyrrolidone carboxyl peptidase (pcp) overexpressed in escherichia coli was purified to homogeneity in less than 12 h using ammonium sulphate precipitation and hydrophobic interaction chromatography. the enzyme, which removes amino-terminal l-pyroglutamic acid from peptides, appears to be a tetramer of 25,200 molecular mass subunits. the protein cross-reacted with polyclonal antibodies raised against pcp from streptococcus pyogenes. the overexpressed enzyme exhibits an absolute ...19921362573
inactivation of bacillus subtilis glutamine synthetase by metal-catalyzed oxidation.instability of bacillus subtilis glutamine synthetase in crude extracts was attributed to site-specific oxidation by a mixed-function oxidation, and not to limited proteolysis by intracellular serine proteases (isp). the crude extract from b. subtilis kn2, which is deficient in three intracellular proteases, inactivated glutamine synthetase similarly to the wild-type strain extract. to understand the structural basis of the functional change, oxidative modification of b. subtilis glutamine synth ...19921363551
[effects of nifuroxazide (ercefuryl), trimethoprim-sulfamethoxazole and bactisubtil in acute diarrhea].the clinical effects of nifuroxasid (n), trimetoprim sulphametoxasol (ts) and bactisubtil (b) on bacillar dysentery and alimentary toxicoinfections in the patients treated at the clinic from january 1984 to the end of december 1989 have been analysed. according to the clinical signs, patients have been divided in ten categories of light, mild and heavy forms. in total, 329 cases of bacillar dysentery and 89 cases of alimentary toxicoinfections have been analysed. the following was established: a ...19911366328
the alpha-amylase gene as a marker for gene cloning: direct screening of recombinant clones.we report the construction and use of a new system for the direct screening of recombinant clones after transformation. the system uses a bacillus subtilis-escherichia coli shuttle vector that carries the b. subtilis structural gene for alpha-amylase. insertion of foreign dna into this gene results in a loss of amylolytic activity in the host cells that can be assayed using a simple and inexpensive staining procedure.19901367421
amperometric determination of ammonium ions with a microbial sensor.a sensor for nh+4 ions has been developed, which consists of immobilized micro-organisms (bacillus subtilis, pseudomonas aeruginosa, trichosporon cutaneum) in combination with an electrochemical transducer. this sensor is based on the measurement of acceleration of respiration after addition of nh+4 in the presence of glucose. the physiological background of this signal and its connection with nh+4 ion uptake and/or metabolism is discussed. the response time of the sensor is about 5-10 s for nh+ ...19901367422
illegitimate recombination in bacillus subtilis: a site-specific mechanism in the formation of plasmid pkbt1.the bacillus subtilis plasmid pkbt1, the product of in vivo rece4-independent recombinal events, contains segments derived from pub110 and the b. subtilis chromosome. to determine whether the pub110 sequence is intact in pkbt1, two 1 kb fragments, each containing a site at which chromosomal and pub110 sequences are joined, were cloned and sequenced. sequencing data revealed that: 1). an intact copy of pub110 is present in pkbt1; 2) the apparent recombination sites were adjacent to the bam hi-gen ...19901367424
the effects of a biosurfactant on oxygen transfer in a cyclone column reactor.a laboratory-scale cyclone column reactor was tested to determine how its oxygen transfer characteristics were affected by surfactants in the liquid medium. the volumetric oxygen transfer coefficient was greatly decreased by small quantities of the synthetic surfactants dodecyltrimethylammonium bromide and sodium dodecylsulfate, and the biosurfactant surfactin produced by bacillus subtilis (atcc 21332). since the gas holdup fraction was generally increased due to foaming, the effectiveness of th ...19901367431
hypersecretion of a cellulase from clostridium thermocellum in bacillus subtilis by induction of chromosomal dna amplification.we have inserted a dna fragment composed of (i) the promoter and the export signal of the bacillus subtilis levansucrase gene; (ii) the sequence encoding the mature part of the clostridium thermocellum endoglucanase a gene in a specific site of the b. subtilis chromosome. the insert was flanked by directly repeated pbr322 sequences of 3.9 kb. plasmid pe194, which has a thermosensitive replication, was integrated adjacent to one of the repeats. when the integrated plasmid was allowed to replicate ...19901367437
subtilisin-catalysed peptide synthesis and transesterification in organic solvents.subtilisin from bacillus subtilis was modified with polyethylene glycol (peg), or adsorbed either on celite or porous glass, or directly used as a suspended powder to catalyse peptide synthesis and transesterification reactions in organic solvents. the rather low yield of peptide synthesis probably resulted from the enzyme tendency to catalyse hydrolysis and transesterification side reactions. the kinetics of transesterification catalysed by peg-subtilisin was consistent with a ping-pong mechani ...19901367438
effect of medium composition on the maintenance of a recombinant plasmid in bacillus subtilis.recombinant plasmid pced3 [confers beta-galactosidase production (lacz+) and kanamycin resistance (kmr)] in bacillus subtilis was found to be both segregationally and structurally unstable. since many solutions to segregational instability are already available, the problem of structural instability was specifically addressed by inclusion of kanamycin in the growth media. culture instability was found to be highest in complex and defined media supporting high growth rates. stabilization over the ...19901367440
recovery of biosurfactants by ultrafiltration.ultrafiltration was used in a one-step method to purify and concentrate biosurfactants--surfactin and rhamnolipids--from culture supernatant fluids. the ability of surfactant molecules to form micelles at concentrations above the critical micelle concentration allows these aggregates to be retained by relatively high molecular weight cut-off membranes. lower molecular weight impurities such as salts, free amino acids, peptides and small proteins are easily removed. various molecular weight cut-o ...19901367444
regulation of biosynthesis of bacilysin by bacillus subtilis.production of the dipeptide antibiotic bacilysin by bacillus subtilis 168 was growth associated and showed no evidence of repression by glucose or sucrose. carbohydrates other than glucose and sucrose yielded lower specific titers of bacilysin. bacilysin production in three such carbon sources (maltose, xylose, ribose) was delayed until growth slowed down. ammonium salts were poor for bacilysin production when used as the sole nitrogen source. when added to the standard medium containing glutama ...19901367485
expression of penicillin g acylase gene from bacillus megaterium atcc 14945 in escherichia coli and bacillus subtilis.penicillin g acylase gene from bacillus megaterium atcc 14945 has been isolated. recombinant escherichia coli clones were screened for clear halo forming activity on the lawn of staphylococcus aureus atcc 6538p using the enzymatic acylating reaction of 7-aminodeacetoxycephalosporanic acid (7-adca) and d-(alpha)-phenylglycine methylester. the gene was contained within a 2.8 kb dna fragment and expressed efficiently when transferred from e. coli to bacillus subtilis. a twenty times greater amount ...19911367491
proteases during purification. 19911367503
a new human growth hormone production process using a recombinant bacillus subtilis strain.we constructed a series of hybrid plasmids which directed the synthesis of different human growth hormone (hgh) precursor sequences in bacillus subtilis. in addition to the 191 amino acids of the hormone, the precursors had in common an amino-terminal extension characterized by the presence of a methionine at position 1 and of the tetrapeptide ile-glu-gly-arg preceding the first residue (phe) of hgh. the sequence between the methionine and the tetrapeptide was specific for each precursor and, be ...19911367506
production processes of recombinant il-1 beta from bacillus subtilis: comparison between intracellular and exocellular expression.the human il-1 beta coding sequence derived from a cdna library was inserted into two different plasmid expression vectors, psm214 and psm308, which promote the synthesis of recombinant products intracellularly and exocellularly, respectively. the hybrid constructs psm261 and psm320 were obtained. bacillus subtilis sms118 was transformed with these plasmids and the recombinant strains were grown in 1 litre bioreactors. different growth conditions were analyzed to optimize yields both in terms of ...19911367530
expression of the bacillus subtilis levanase gene in escherichia coli and saccharomyces cerevisiae.the gene coding for the inulin hydrolyzing enzyme levanase which was previously cloned from bacillus subtilis was fused to the tac-promoter. overexpression in escherichia coli resulted in high amounts of intracellularly produced levanase (up to 20 u mg-1). after removal of the bacterial 5' sequences, the levanase gene was also cloned into a yeast expression vector based on the pgk-promoter. clones containing the intact levanase gene including the bacterial signal sequence gave rise to synthesis ...19911367531
production of pectin methylesterase from erwinia chrysanthemi b374 in bacillus subtilis.the gene coding for pectin methylesterase (pme) of erwinia chrysanthemi b374 (pme) was cloned by a polymerase chain reaction. the pme gene was expressed in bacillus subtilis using a secretion vector based on the promoter and signal sequence of the alpha-amylase gene from b. amyloliquefaciens. the cultivation of b. subtilis cells carrying the cloned pme resulted in efficient secretion of pme into the culture medium based on enzymatic and sodium dodecyl sulphate-poly-acrylamide gel electrophoresis ...19911367534
continuous regeneration of nad(h) covalently bound to a cysteine genetically engineered into glucose dehydrogenase.we introduced a cysteine residue on the surface of glucose dehydrogenase from bacillus subtilis using site-directed mutagenesis. to this mutant, an nad-analogue was covalently attached by a disulphide bridge so that it was active intramolecularly. the glucose dehydrogenase-cys44-nad complex, which contained one reactive nad molecule per subunit of glucose dehydrogenase, was operated together with lactate dehydrogenase in a coupled enzymatic regeneration of nad(h) in a hollow fiber reactor. l-lac ...19911367536
proteolytic activities in bacillus.major advances have been made in understanding the regulation of expression of bacillus subtilis protease genes. a phosphorelay mechanism as well as a two-component regulatory system allow conditions of the growth medium to be transmitted to the gene level resulting in expression of extracellular protease genes.19911367717
on the safety of bacillus subtilis and b. amyloliquefaciens: a review. 19911367772
production of heterologous proteins in bacillus subtilis: the effect of the joint between signal sequence and mature protein on yield.we have previously made a set of dna constructs by fusing the mature part of bacillus licheniformis alpha-amylase with the signal sequence of b. amyloliquefaciens alpha-amylase at various distances from the signal sequence cleavage site. we observed that the level of alpha-amylase production in b. subtilis depended strongly on the distance of the junction from the signal sequence cleavage site, with quite a sharp optimum distance. to test whether the effect is limited to the pair of alpha-amylas ...19911367777
processing of the prepropeptide portions of the bacillus amyloliquefaciens neutral protease fused to bacillus subtilis alpha-amylase and human growth hormone during secretion in bacillus subtilis.a set of nested 3'-terminal deletions of the prepropeptide of the bacillus amyloliquefaciens neutral protease gene was constructed. alpha-amylase and human growth hormone were secreted using these truncated genes in bacillus subtilis. the level of the secreted alpha-amylase varied with the region for the truncated prepropeptide contained in the fusion gene but was independent of its length. even though length of the prepropeptide varied, the mobilities of secreted alpha-amylases were the same as ...19921367948
cloning and expression of the lysostaphin gene in bacillus subtilis and lactobacillus casei.the lysostaphin structural gene was cloned in bacillus subtilis dsm402 and in lactobacillus casei 102s. the gene was expressed in both organisms and active lysostaphin was released into the medium. lysostaphin produced by these organisms induced lysis of growing and heat inactivated staphylococci. expression in a protective starter organism is a prerequisite to produce lysostaphin in situ in fermenting foods and hence, to reduce the hygienical risk of staphylococcal food poisoning.19921367977
secretion of correctly processed and folded pancreatic secretory trypsin inhibitor by bacillus subtilis.we constructed a plasmid, designated pnpp126, containing a dna sequence encoding a fusion protein composed of bacillus amyloliquefaciens neutral protease prepeptide (signal peptide) and human pancreatic secretory trypsin inhibitor (hpsti), where the mature hpsti is accurately fused to the 3'-terminal of the prepeptide coding region. it was observed that the strain bacillus subtilis mt600 harboring pnpp126 could secrete a trypsin inhibitory activity into the culture medium. the n-terminal amino a ...19921368060
high expression vectors for the production of recombinant single-chain urinary plasminogen activator from escherichia coli.an expression cassette containing a synonymous gene for human single-chain urokinase-type plasminogen activator (rscu-pa) 5'-flanked by a trp promoter and the shine-dalgarno sequence of the xyl a operon of bacillus subtilis and terminated by the terminators trp a and tn10 was constructed and inserted into a pbr322 derivative to yield pbf160. when compared to puk54 trp 207-1 containing the natural scu-pa gene without the shine-dalgarno sequence and terminator, the expression efficiency of pbf160 ...19921368067
ph-dependent flagella formation by facultative alkaliphilic bacillus sp. c-125.a facultative alkaliphilic strain of bacillus sp. c-125 grown at alkaline ph had many sinuous peritrichous flagella and was highly motile. however, most of the cells grown initially at ph 7 were non-motile and possessed few straight flagella. the amount of flagellin was low when the organism was grown at ph 7, suggesting that non-motility is due to poor synthesis of flagellin. the molecular mass of the flagellin was 37 kda and the isoelectric point was ph 5.0. the amino acid composition of the f ...19921368134
purification and characterization of an antibiotic substance produced from rhizopus oligosporus ifo 8631.we obtained a purified antibiotic protein from the submerged cultivation broth of rhizopus oligosporus ifo 8631 by using cm-cellulofine chromatography and hplc. the antibiotic did not show a broad spectrum of activity, but it was very active against some of the bacillus species, especially against bacillus subtillis (b. natto) at a very low concentration (less than 1 ppm). it also showed activity against other gram-positive bacteria, including staphylococcus aureus and streptococcus cremoris. th ...19921368137
construction of a novel tetracycline resistance gene cassette useful as a marker on the bacillus subtilis chromosome. 19921368214
expression of cloned homologous fermentative genes in clostridium acetobutylicum atcc 824.we have previously cloned the acetone-formation pathway gene, encoding acetoacetate decarboxylase (adc), and butyrate-formation pathway gene, encoding phosphotransbutyrylase (ptb), of clostridium acetobutylicum atcc 824 in escherichia coli. here we report their subcloning in bacillus subtilis and transfer to strain atcc 824 via electrotransformation, where the corresponding enzyme activities were expressed at elevated levels, using pfnk1, a new b. subtilis/c. acetobutylicum shuttle vector. plasm ...19921368230
protein secretion in gram-positive bacteria.gram-positive bacteria often secrete large amounts of proteins into the surrounding medium. this feature makes them attractive as hosts for the industrial production of extracellular enzymes. compared to escherichia coli, relatively little is known about the mechanism of protein secretion in these organisms. however, the recent identification of bacillus subtilis genes whose gene products are highly homologous to some of the sec (secretion) proteins of e. coli strongly suggests that important pr ...19921368245
maturation of an nh2-terminally extended thermostable alpha-amylase in bacillus subtilis: a possible mechanism examined by in vitro experiments.an artificially inserted extra peptide (21 amino acid peptide) between the b. subtilis alpha-amylase signal peptide and the mature thermostable alpha-amylase was completely cleaved by b. subtilis alkaline protease in vitro. the cleavage to form a mature enzyme was observed between ph 7.5 and 10, but not between ph 6.0 and 6.5, although a similar protease activity toward azocall was observed between ph 6.0 and 7.5. to analyze the effects of ph on the cleavage, cd spectra at ph 6, 8, and 11 of the ...19921368250
nucleotide sequence of the gene for an alkaline endoglucanase from an alkalophilic bacillus and its expression in escherichia coli and bacillus subtilis.the gene for an alkaline endoglucanase from the alkalophilic bacillus sp. ksm-64 was cloned into the hindiii site of pbr322 and expressed in escherichia coli hb101. the nucleotide sequence of a 4.1-kb region of the hindiii insert had two open reading frames, orf-1 and orf-2. the protein deduced from orf-1 was composed of 244 amino acids with an m(r) of 27,865. subcloning analysis proved that the alkaline endoglucanase was encoded by orf-2 (822 amino acids with an m(r) of 91,040). upstream from o ...19921368251
physiological and genetic strategies for enhanced subtilisin production by bacillus subtilis.defined minimal media conditions were used to assess and subsequently enhance the production of subtilisin by genetically characterized bacillus subtilis strains. subtilisin production was initiated by the exhaustion or limitation of ammonium in batch and fed-batch cultures. expression of the subtilisin gene (apre) was monitored with a chromosomal apre::lacz gene fusion. the beta-galactosidase production driven by this fusion reflected subtilisin accumulation in the culture medium. subtilisin ge ...19921368258
sequence analysis of replication origin of plasmid pls11 of bacillus subtilis ifo 3022.the structure of a 1.6-kb sphi-hindiii dna sequence necessary and sufficient for the replication of a 8.6-kb plasmid pls11 of bacillus subtilis ifo 3022, which is responsible for gamma-polyglutamate production, has been characterized by using a trimethoprim (tmp)-resistance gene derived form b. subtilis ttk24 chromosomal dna as a selective marker. the 1.6-kb dna sequence contains a rep gene encoding the protein (333 amino acids) essential for initiation of replication and a possible origin of re ...19921368298
bacillus subtilis and its relatives: molecular biological and industrial workhorses.the non-pathogenic bacterium bacillus subtilis, since its first reported genetic transformation in 1959, has become a model system for the study of many aspects of the biochemistry, genetics and physiology of gram-positive bacteria, and particularly of sporulation and associated metabolism. extensive knowledge of the molecular biology of b. subtilis has led to the recent development of this bacterium as a host for the industrial production of heterologous proteins. although difficulties have bee ...19921368322
effect of parb on plasmid stability and gene expression in xanthomonas campestris.the stabilization locus parb was subcloned into the broad host range plasmid pap2, which contains the alpha-amylase gene from bacillus subtilis, and introduced into xanthomonas campestris pv campestris and x.c.pv manihotis. analysis of the stability of plasmid pap2 (parb-) and pap23 (parb+) showed that the parb locus decreased significantly the plasmid loss rate mainly by x.c.pv campestris. the lower efficiency of stabilization in x.c.pv manihotis was probably due to the incompatibility system b ...19921368368
diffusion of proteases in calcium alginate beads.diffusion of proteases from bacillus subtilis and serratia marcescens within calcium alginate beads has been assayed, and the experimental data fitted into a mathematical model for diffusion into a finite volume liquid medium. values of effective diffusivity were calculated for the studied proteases and compared with the available data in the literature for molecules of lower molecular weight.19921368429
structure of di-o-alpha-maltosyl cyclodextrins produced from alpha-maltosylfluoride and cyclodextrins.the structures of di-o-alpha-maltosyl beta-cyclodextrins ((g2)2-beta-cds), which were produced from alpha-maltosylfluoride (alpha-g2f) and cyclodextrin (cd) by the transfer action of debranching enzymes, were examined by the enzymic method using bacillus subtilis saccharifying alpha-amylase (bsa). (g2)2-beta-cd was converted to (g1)2-beta-cd by treatment with glucoamylase before the examination. bsa completely hydrolyzed (g1)2-beta-cd to produce glucose, 6(3)-o-alpha-glucosylmaltotriose, and 6(3 ...19901368598
construction of a bacillus subtilis strain deficient in three proteases. 19901368619
promotive and inhibitory effects of raw starch adsorbable fragments from pancreatic alpha-amylase on enzymatic digestions of raw starch.the enzymatically inactive but raw-starch-adsorbable peptide fragments designated as gp-pan p and gp-pan i were obtained from a tryptic digest of heat-inactivated hog pancreatic alpha-amylase. these two glycopeptide fragments were purified with sephadex g-75, deae-sephadex a-50, and hplc and were found to be homogeneous on disc gel electrophoresis. gp-pan p and i had molecular weights of 20,000 and 30,000 with sds-page, carbohydrate contents of 10% and 7%, n-terminal amino acids gly-trp and ala- ...19911368657
xylose (glucose) isomerase gene from the thermophile clostridium thermohydrosulfuricum; cloning, sequencing, and expression in escherichia coli.the xylose isomerase gene from the thermophile clostridium thermohydrosulfuricum has been cloned, using a fragment of the bacillus subtilis gene as a probe. the complete nucleotide sequence of the gene was analyzed. c. thermohydrosulfuricum is the most thermophilic organism from which a xylose isomerase gene has been cloned and characterized. comparison with amino acid sequences from other xylose isomerases showed that amino acids involved in substrate binding and isomerization are well conserve ...19911368665
cloning, sequencing, and characterization of the intracellular invertase gene from zymomonas mobilis.the structural gene for the intracellular invertase e1 of zymomonas mobilis strain z6c was cloned in a 2.25-kb dna fragment on push11, and expressed in escherichia coli hb101. the enzyme produced by the e. coli carrying push11 was purified about 1,122 fold to homogenicity with a yield of 4%. the molecular weight and substrate specificity of the enzyme were identical with those of the intracellular invertase e1 from z. mobilis. the nucleotides of the cloned dna were sequenced; they included an op ...19911368686
characterization and structure of the cellulase gene of bacillus subtilis bse616.the bacillus subtilis carboxymethyl cellulase (cmcase) gene originally cloned on a 3.2-kb psti dna fragment has been localized in a 1.5-kb sau3ai fragment by a series of subclonings into plasmid puc19. during the process the promoter region and shine-dalgarno (sd) sequence were deleted, but the 1.5-kb insert was shown to direct the synthesis of cmcase in scherichia coli to a high level, probably with the aid of lac promoter. we analyzed the complete nucleotide sequence of the cmcase gene. the cm ...19911368694
molecular cloning and analysis of nucleotide sequence of the bacillus subtilis lysa gene region using b. subtilis phage vectors and a multi-copy plasmid, pub110.a 3.8-kb ecori-fragment containing the lysa gene [diaminopimelate (dap) decarboxylase] of bacillus subtilis has been cloned into b. subtilis phage phi 105 and its nucleotides sequenced. the nucleotide sequence of a 3,762 bp stretch contained three open reading frames (orf1, orf2, and orf3) in one orientation and another open reading frame (orf4) in the opposite orientation. orf2 coded for the lysa gene based on the complementation of a b. subtilis lys auxotroph and on the fact that the predicted ...19911368705
transfer reaction catalyzed by exo-beta-1,4-galactanase from bacillus subtilis.a transfer reaction catalyzed by an exo-beta-1,4-galactanase from bacillus subtilis was studied. the enzyme had a broad acceptor specificity and transferred galactobiosyl residues to acceptors such as various alcohols, including hydroxy benzenes and saccharides. transfer products of glycerol formed by the enzyme were compared with those formed by escherichia coli beta-galactosidase and by penicillium citrinum endo-galactanase. e. coli enzyme transferred 90% of galactose residues to the primary h ...19911368730
dye-sensitized photooxidation of neutral protease from bacillus subtilis var. amylosacchariticus: assignment of histidine residue oxidized.the neutral protease of bacillus subtilis var. amylosacchariticus was photooxidized in the presence of methylene blue, by which treatment the enzyme was rapidly inactivated. the inactive enzyme was digested with endoproteinase asp-n, the resultant peptides were separated by hplc, and their amino acid sequences were compared with those obtained from the unmodified enzyme. of four peptides that contained histidine residues, only the recovery of one peptide was found to be decreased by the photooxi ...19911368731
use of a triple protease-deficient mutant of bacillus subtilis as a host for secretion of a b. subtilis cellulase and tem beta-lactamase.the mature portion of the tem beta-lactamase (bla) gene (bla) derived from pbr322 was fused with the promoter and signal region of bacillus subtilis cellulase (bsc) gene (bsc), and the productivity was compared with that of the cloned native bsc gene, using a wild-type b. subtilis strain and a strain deficient in three proteases (i.e., extracellular serine protease, extracellular neutral protease, and the major intracellular serine protease) as hosts. the effects of the sen, sacq, and prtr genes ...19911368741
chemical modification of neutral protease from bacillus subtilis var. amylosacchariticus: assignment of tyrosyl residues iodinated.the neutral protease of bacillus subtilis var. amylosacchariticus (b. amylosacchariticus) was iodinated with a 25-fold molar excess of iodine at ph 9.4 for 3 min at 0 degree c, by which treatment the proteolytic activity toward casein was markedly reduced, while the hydrolytic activity toward an n-blocked peptide substrate was rather increased. the modified enzyme was digested with staphylococcus aureus v8 protease at ph 8.0 and the amino acid sequences of resultant peptides were compared with t ...19911368747
production of cyclomaltodextrin glucanotransferase of bacillus circulans var. alkalophilus atcc21783 in b. subtilis.the cyclomaltodextrin glucanotransferase (cgtase, e.c. 2.4.1.19) gene from an alkalophilic bacillus circulans var. alkalophilus atcc21783 was cloned into escherichia coli and b. subtilis. when cloned from e. coli to b. subtilis, the entire insert containing the cgtase gene was, depending on the plasmid construction, either unstable or the recombinant b. subtilis did not secrete the enzyme in significant amounts. to achieve efficient enzyme production in b. subtilis, the gene was placed under the ...19921368772
purification of a new extracellular 90-kda serine proteinase with isoelectric point of 3.9 from bacillus subtilis (natto) and elucidation of its distinct mode of action.a new extracellular 90-kda serine proteinase with an isoelectric point (pi) of 3.9 was purified from bicillus subtilis (natto). microheterogeneity was detected in the 50-kda protease of bacillopeptidase f with pi 4.4 reported previously by wu et al. and the sequence for the first 25 amino acids in the internal region of the enzyme was analyzed: atdgvewnvdqidapkawalgydga. the cleavage sites in the oxidized b-chain of insulin by the proteinase were cyso3h7-gly8, val12-glu13, try16-leu17, and phe25 ...19921368833
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