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[the effect of growth media on recovery of test microorganisms after exposure to saturated steam under pressure].the aim of this study was to find out which growth media give the best condition for the development of test bacteria after exposure to saturated steam under pressure. the test organisms were strains of bacillus subtilis nctc 3610 and bacillus stearothermophilus nctc 8923. the test prepared from spore suspension were exposed to saturated steam under pressure 0.2 atn-b.subtilis, and 0.7 atn-b. stearothermophilus with various length of exposure /sublethal conditions/. after the exposure the tests ...19969102803
effect of high-power microwave on indicator bacteria for sterilization.according to the superiority sterilization, a specially-designed microwave disinfector using high-power microwave energy was made. a series of sterilizing experiments have been made to determine the effect of microwave energy on several typical indicator bacteria such as bacillus subtilis var. niger, bacillus stearothermophilus, bacillus pumilus e601, staphylococcus aureus, bacillus cereus. under the conditions of different sterilization duration and unequal intensity of microwave power irradiat ...19969216147
purification and characterization of dna helicase bsth1 from bacillus stearothermophilus.we have partially purified a dna helicase bsth1 from bacillus stearothermophilus through polymin p precipitation, ammonia sulfate precipitation and column chromatographic steps with pheny1-sepharose, deae-cellulose, phosphocellulose, fplc mono q and superose 12. bsth1 possesses a dna-dependent atpase activity in the presence of mg(2+). the atpase activity of bsth1 is differentially stimulated by the presence of different types of nucleic acids. bsth1 has an optimal atpase activity at 55 degrees. ...199612232591
affinity precipitation and site-specific immobilization of proteins carrying polyhistidine tails.proteins carrying genetically attached polyhistidine tails have been purified using affinity precipitation with metal chelates. dna fragments encoding four or five histidine residues have been genetically fused to the oligomeric enzymes lactate dehydrogenase (bacillus stearothermophilus), beta-glucoronidase (escherichia coli), and galactose dehydrogenase (pseudomonas fluorescens) as well as to the monomeric protein a (staphylococcus aureus). the chimeric genes were subsequently expressed in e. c ...199618624332
study of stability of recombinant plasmids during the continuous culture of bacillus stearothermophilus nub3621 in nonselective medium.the optimal culture conditions for bacillus stearothermophilus nub3621 (bgsc 9a5) in chemostat were studied. the results obtained showed that the optimal culture conditions in terms of biomass concentration and maximum growth rate were 65 degrees c, ph 6.8 to 7.2. dissolved oxygen became growth limiting at po(2) levels below 10%. furthermore, this strain was transformed with three new hybrid vectors (ppam2, ppch2, or pply2) constructed by cloning in prp9, a plasmid based on the thermophilic repl ...199718634046
mass production of thermostable d-hydantoinase by batch culture of recombinant escherichia coli with a constitutive expression system.d-hydantoinase is an industrial enzyme widely used for the synthesis of optically active d-amino acids. a gene encoding thermostable d-hydantoinase of bacillus stearothermophilus sd-1 has previously been cloned and constitutively expressed by its native promoter in escherichia coli xl1-blue (lee et al., 1996b). in this work, we attempted mass production of the d-hydantoinase by batch culture of the recombinant e. coli using glycerol as a carbon source. the plasmid content in cells increased in p ...199718642247
crystallization and preliminary x-ray analysis of the thermostable alkaline-tolerant xylanase from bacillus stearothermophilus t-6.the extracellular thermostable xylanase (xt-6) produced by the thermophilic bacterium bacillus stearothermophilus t-6 was shown to bleach pulp optimally at ph 9 and 338 k, and was successfully used in a large-scale biobleaching mill trial. the xylanase gene was cloned and sequenced. the mature enzyme consists of 379 amino acids with a calculated molecular weight of 43,808 and pi of 9.0. crystallographic studies of xt-6 were initiated to study the mechanism of catalysis as well as to provide a st ...199715299894
purification, crystallization and preliminary x-ray analysis of the m.bseci dna methyltransferase from bacillus stearothermophilus.the dna methyltransferase m.bsec1 from b. stearothermophilus methylates the n6 atom of the 3' adenine in the sequence 5'-atcgat-3'. the 579-residue protein has been isolated and crystallized using seeding and microdialysis techniques. the crystals are monoclinic, space group p2(1) with cell dimensions a = 53.7, b = 85.7, c = 151.8 a and beta = 95.1 degrees, two molecules in the asymmetric unit and diffract to at least 2.5 a resolution.199715299919
mathematical models for the effects of ph, temperature, and sodium chloride on the growth of bacillus stearothermophilus in salty carrots.estimating the shelf life and safety of any food product is an important part of food product development. predictive food microbiology reduces the time and expense associated with conventional challenge and shelf life testing. the purpose of this study was to characterize and model germination, outgrowth, and lag (gol) time and the exponential growth rate (egr) of bacillus stearothermophilus in salty carrot medium (scm) as a function of ph, temperature, and nacl concentration. b. stearothermoph ...199716535566
stabilization of enzymes against thermal stress and freeze-drying by mannosylglycerate.2-o-(beta)-mannosylglycerate, a solute that accumulates in some (hyper)thermophilic organisms, was purified from pyrococcus furiosus cells, and its effect on enzyme stabilization in vitro was assessed. enzymes from hyperthermophilic, thermophilic, and mesophilic sources were examined. the thermostabilities of alcohol dehydrogenases from p. furiosus and bacillus stearothermophilus and of glutamate dehydrogenases from thermotoga maritima and clostridium difficile were improved to a significant ext ...199716535713
purification and characterization of dna helicase bsth2 from bacillus stearothermophilus.in the purification of dna helicase bsth1 we have partially purified the second dna helicase bsth2 from bacillus stearothermophilus through polymin p precipitation, ammonia sulfate precipitation and chromatographic steps with deae-cellulose, phosphocellulose, blue-sepharose, fplc superose 12, mono q and second mono q. the atpase activity of bsth2 depends on mg(2+) and is differentially stimulated by different types of nucleic acids. bsth2 has a maximal atpase activity at 55 degrees. the atpase a ...199712215800
growth-promoting effect of carbon material upon bacterial cells propagating through a distance.carbon material such as graphite and activated charcoal, but not diamond, causes the promotion of growth of certain bacteria under ordinarily non-permissive stress conditions over a distance of several centimeters. bacillus carboniphilus under the stress of a high kcl concentration and high temperature responded to this remote effect of carbon material with enhanced growth, and thermophile bacterium bacillus stearothermophilus responded similarly yet moderately under the stress of low temperatur ...199712501323
antibiotic residues and prevalence of mastitis pathogen isolation in heifers during early lactation following prepartum antibiotic therapy.the present study was conducted to determine if antibiotic treatment of heifer mammary glands earlier in the prepartum period reduced the occurrence of residues in milk without compromising efficacy in treatment of intramammary infections. heifers were assigned randomly to two groups: 1. untreated negative control (n = 42); and 2. intramammary infusion of 200 mg cephapirin sodium (n = 40) 14 days prior to expected calving. mammary secretions were collected before treatment and during early lacta ...19979230672
primary structure, sequence analysis, and expression of the thermostable d-hydantoinase from bacillus stearothermophilus sd1.the gene coding for the thermostable d-hydantoinase from the thermophilic bacterium bacillus stearothermophilus sd1 was cloned and its nucleotide sequence was completely determined. the d-hydantoinase protein showed considerable amino acid sequence homology (20-28%) with other hydantoinases and functionally related allantoinases and dihydroorotases. strikingly the sequence of the enzyme from b. stearothermophilus sd1 exhibited greater than 89% identity with hydantoinases from thermophilic bacter ...19979236771
characterization of a gene cluster for glycogen biosynthesis and a heterotetrameric adp-glucose pyrophosphorylase from bacillus stearothermophilus.a chromosomal region of bacillus stearothermophilus trbe14 which contains genes for glycogen synthesis was cloned and sequenced. this region includes five open reading frames (glgbcdap). it has already been demonstrated that glgb encodes branching enzyme (ec 2.4.1.18 [h. takata et al., appl. environ. microbiol. 60:3096-3104, 1994]). the putative glgc (387 amino acids [aa]) and glgd (343 aa) proteins are homologous to bacterial adp-glucose pyrophosphorylase (agp [ec 2.7.7.27]): the sequences shar ...19979244254
[study on transposition behavior of is5376 in bacillus stearothermophilus].is5376 and is5377 are two transposable elements discovered in bacillus stearothermophilus. analysis of random samples revealed that the frequency of transposition of is5376 from cu21 chromosome to plasmids pfdc5 and pfdc12 was much higher at 65 degrees c than that at 48 degrees c while that of is5377 was very low at both 48 degrees c and 65 degrees c. the exact nature of the temperature effect is obscure at present from evidences obtained so far it is concluded that this is a consequence of the ...19979254976
further studies on thermal denaturation of pyruvate dehydrogenase complex from bacillus stearothermophilus.thermally induced changes in pyruvate dehydrogenase complex (pdc) from b. stearothermophilus were examined mainly at temperatures from 60 degrees to 70 degrees c. accompanied by inactivation of pyruvate decarboxylase, light scattering decreased, and ans fluorescence increased. these changes including the inactivation were approximately first-order reactions, and the values of rate constants were greatly dependent on temperature. chromatographic studies showed that any polypeptides were in associ ...19979255976
synthesis of glycosyl-trehaloses by cyclomaltodextrin glucanotransferase through the transglycosylation reaction.cyclomaltodextrin glucanotransferase from bacillus stearothermophilus produced a series of glycosyl-trehaloses through the transglycosylation reaction with cyclomaltohexaose as the glycosyl donor and trehalose as its acceptor. after beta-amylase treatment, five species of glycosyl-trehaloses were isolated by column chromatography. after chemical and enzymatic analyses, it was concluded that these oligosaccharides were alpha-maltosyl alpha-d-glucopyranoside, alpha-maltotriosyl alpha-d-glucopyrano ...19979255978
nucleotide sequence and characterization of the cryptic bacillus thuringiensis plasmid pgi3 reveal a new family of rolling circle replicons.the complete nucleotide sequence of plasmid pgi3 from bacillus thuringiensis subsp. thuringiensis h1.1. was obtained. although this 11,365-bp molecule contained at least 11 putative open reading frames (orfs), extensive database searches did not reveal any homologous sequences with the exception of orf6, which displayed similarity to the largest orf of pstk1, a 1,883-bp cryptic plasmid isolated from bacillus stearothermophilus. deletion analysis to determine the pgi3 minimal replicon revealed th ...19979260939
the use of two or more microorganisms versus one microorganism in the carrier materials for biological indicators.specification for the preparation of a biological indicator (bi) using two or more microorganisms in the carrier material were deleted from the current iso 11138 series and working draft 14161 because it was assumed that the resistances of the individual microorganisms would be affected by interference from the other microorganisms. this assumption is speculative only, and has not been supported by experimental evidence. to test its validity, the author carried out an experiment to determine the ...19979262838
sequencing and phylogenetic analysis of the spoiia operon from diverse bacillus and paenibacillus species.in order to clone the spoiia operon from three different bacillus and paenibacillus species, we designed two sets of pcr primers based on three previously published bacillus spoiia sequences. one set of primers corresponded to the c-terminal region of spoiiab and a region near the middle of spoiiac. these primers were used to amplify the corresponding region of spoiia from bacillus stearothermophilus and paenibacillus polymyxa (previously called bacillus polymyxa [see ash, c., priest, f.g., coll ...19979266669
cloning and sequencing of the hrca gene of bacillus stearothermophilus.we report on cloning and sequencing of a 2.0-kb pcr fragment of chromosomal dna from thermophilic bacillus stearothermophilus carrying the complete hrca gene. in addition, this amplicon contains the 3' end of an open reading frame exhibiting significant homology to the hemn gene of bacillus subtilis (bs) and other bacterial species. the hrca gene could complement an bs hrca deletion mutant by repressing expression of class i heat shock (hs) genes. furthermore, we could show that the hrca protein ...19979266682
analysis of the bacillus subtilis genome: cloning and nucleotide sequence of a 62 kb region between 275 degrees (rrnb) and 284 degrees (pai).in the framework of the international project aimed at the sequencing of the bacillus subtilis genome, five dna fragments in the region between rrnb (275 degrees) and pai (284 degrees) were cloned by inverse and combinatorial long-range pcr and their nucleotide sequences were determined and analysed. together these sequences constituted a contig of 62229 bp. on the basis of the position of not1 and stil restriction sites, the orientation and order of known genetic markers was determined to be pa ...19979274030
molecular biology of s-layers.in this chapter we report on the molecular biology of crystalline surface layers of different bacterial groups. the limited information indicates that there are many variations on a common theme. sequence variety, antigenic diversity, gene expression, rearrangements, influence of environmental factors and applied aspects are addressed. there is considerable variety in the s-layer composition, which was elucidated by sequence analysis of the corresponding genes. in corynebacterium glutamicum one ...19979276928
functions of s-layers.although s-layers are being increasingly identified on bacteria and archaea, it is enigmatic that in most cases s-layer function continues to elude us. in a few instances, s-layers have been shown to be virulence factors on pathogens (e.g. campylobacter fetus ssp. fetus and aeromonas salmonicida), protective against bdellovibrio, a depository for surface-exposed enzymes (e.g. bacillus stearothermophilus), shape-determining agents (e.g. thermoproteus tenax) and nucleation factors for fine-grain m ...19979276929
the catalytic domain of dihydrolipoyl acetyltransferase from the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus. expression, purification and reversible denaturation.a sub-gene encoding the catalytic (acetyltransferase) domain (e2pcd) comprising residues 173-427 of the dihydrolipoyl acetyltransferase (e2p) chain of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus was expressed in escherichia coli. the product assembled to form the characteristic icosahedral (60-mer) core structure with full catalytic activity. the km values for dihydrolipoamide and acetyl-coa were 1.2 mm and 13 microm, respectively. dissociation of the icosahedra ...19979280309
the highly thermostable arginine repressor of bacillus stearothermophilus: gene cloning and repressor-operator interactions.we report here the cloning of the arginine repressor gene argr of bacillus stearothermophilus and the characterization and purification to homogeneity of its product. the deduced amino acid sequence of the 16.8-kda argr subunit shares 72% identity with its mesophilic homologue ahrc of bacilus subtilis. sequence analysis of b. stearothermophilus argr and comparisons with mesophilic arginine repressors suggest that the thermostable repressor comprises an n-terminal dna-binding and a c-terminal oli ...19979282750
thermal conversion from low- to high-activity forms of catalase i from bacillus stearothermophilus.catalase i from bacillus stearothermophilus has the interesting property of increasing its enzyme activity on heating. it was confirmed that after heating at 70 degrees c for 10 min or 65 degrees c for 20 min, almost all the enzyme molecules were converted irreversibly to the activated form. the increase in kcat from 1400 to 3930 s-1 and the decrease in km for h2o2 from 4.4 to 2.7 mm by heat activation indicate changes in the kinetic property of the enzyme molecule. therefore, it follows that ca ...19979287297
bacillus stearothermophilus as a model to evaluate membrane toxicity of a lipophilic environmental pollutant (ddt).the thermophilic eubacterium bacillus stearothermophilus has been used as a model system to identify ddt-promoted events in biological membranes putatively related with the insecticide toxicity. two strategies have been approached: a) bacterial growth and viability were followed and the effects of ddt (2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane) determined; b) biophysical studies with fluorescent probes were performed to elucidate the effects of ddt on the organization of the membrane lipid b ...19979294237
gene organisation and regulatory sequences in the sucrose utilisation cluster of bacillus stearothermophilus nub36.the nucleotide sequence of the surp and surt genes in a sucrose-utilisation cluster cloned from bacillus stearothermophilus nub36 was determined. the surp gene encoded a protein of 466 amino acid residues and shared 60-62% amino acid identity with the sucrose-specific enzyme ii components of the phosphotransferase system of bacillus subtilis, salmonella typhimurium and klebsiella pneumoniae. surp, like other sucrose eiis, lacked the hydrophilic domain containing the first (iia) phosphorylation s ...19979305764
bacterial killing ability of 10% ethylene oxide plus 90% hydrochlorofluorocarbon sterilizing gas.to use a serum and salt challenge in narrow-lumen carriers to evaluate a 10% ethylene oxide plus 90% hydrochlorofluorocarbon (eo-hcfc) sterilant mixture in a retrofitted 12/88 sterilizer as an alternative to the banned chlorofluorocarbon-ethylene oxide (eo) sterilant mixture.19979309437
carbohydrate protection of enzyme structure and function against guanidinium chloride treatment depends on the nature of carbohydrate and enzyme.baker's yeast cells accumulate osmolytes as a response to several stress conditions such as high-temperature and low-temperature shifts, dehydration, or osmotic stress. one of the major osmolytes that accumulates is trehalose, which plays an essential role affecting the survival of yeast at the time of stress. in this report, we show that trehalose efficiently protects the function and the structure of two yeast cytosolic enzymes against chemical denaturation by guanidinium chloride. other sugar ...19979310355
cloning of the bsshii restriction-modification system in escherichia coli : bsshii methyltransferase contains circularly permuted cytosine-5 methyltransferase motifs.bsshii restriction endonuclease cleaves 5'-gcgcgc-3' on double-stranded dna between the first and second bases to generate a four base 5'overhang. bsshii restriction endonuclease was purified from the native bacillus stearothermophilus h3 cells and its n-terminal amino acid sequence was determined. degenerate pcr primers were used to amplify the first 20 codons of the bsshii restriction endonuclease gene. the bsshii restriction endonuclease gene (bsshiir) and the cognate bsshii methyltransferase ...19979321648
partition of ddt and dde into membranes and extracted lipids of bacillus stearothermophilus. 19979323216
correlation of the enzyme activities of bacillus stearothermophilus lactate dehydrogenase on three substrates with the results of molecular dynamics/energy minimization conformational searching.current methods for reengineering enzyme substrate specificities rely heavily on the use of static x-ray crystallographic models. in this article we detail the use of a molecular mechanics approach for suggesting regions of bacillus stearothermophilus l-lactate dehydrogenase (ec 1.1.1.27) involved in substrate specificity, and hence areas of interest for protein engineers. the approach combines molecular dynamics with energy minimization (md/em) to search the conformational space available to a ...19979329087
ribosomal protein s7: a new rna-binding motif with structural similarities to a dna architectural factor.the ribosome is a ribonucleoprotein complex which performs the crucial function of protein biosynthesis. its role is to decode mrnas within the cell and to synthesize the corresponding proteins. ribosomal protein s7 is located at the head of the small (30s) subunit of the ribosome and faces into the decoding centre. s7 is one of the primary 16s rrna-binding proteins responsible for initiating the assembly of the head of the 30s subunit. in addition, s7 has been shown to be the major protein comp ...19979331423
rapid identification of heterotrophic, thermophilic, spore-forming bacteria isolated from hot composts.the restriction enzyme profiles of 16s ribosomal dnas (rdnas) amplified by pcr from thermophilic heterotrophic bacterial strains isolated from composts were compared with those of reference strains. this allowed us to assign all but 1 of 16 strains to four different bacillus species (namely, bacillus stearothermophilus, bacillus pallidus, bacillus thermoglucosidasius, and "bacillus thermodenitrificans"). this study showed that pcr restriction analysis of 16s rdna contributes to rapid and reliabl ...19979336936
general equation of steady-state enzyme kinetics using net rate constants and its applicaiton to the kinetic analysis of catalase reaction.the steady-state velocity equation is derived for the general reaction scheme containing n kinds of enzyme species connected by a network of reversible reaction steps. the general equation is represented by the net rate constants as well as the true rate constants of the individual reaction steps. using a general equation, equations for simpler schemes can easily be derived. furthermore, the velocity equation expressed by net rate constants is useful for understanding the dynamic state of any re ...19979344734
mutational analysis of a surface area that is critical for the thermal stability of thermolysin-like proteases.site-directed mutagenesis was used to assess the contribution of individual residues and a bound calcium in the 55-69 region of the thermolysin-like protease of bacillus stearothermophilus (tlp-ste) to thermal stability. the importance of the 55-69 region was reflected by finding that almost all mutations had drastic effects on stability. these effects (both stabilizing and destabilizing) were obtained by mutations affecting main chain flexibility, as well as by mutations affecting the interacti ...19979346299
a novel levansucrase-levanase gene cluster in bacillus stearothermophilus atcc12980.a 3.1 kb fragment of bacillus stearothermophilus atcc12980 dna permitted growth of escherichia coli on sucrose. the fragment encoded genes for a levansucrase (surb) and also a levanase (surc). surb and surc shared 97% and 43% amino acid identity with the corresponding sacb and sacc proteins of bacillus subtilis, whose sacb and sacc genes are organised very differently.19979349714
crystallization and preliminary x-ray crystallographic study of the ribosomal protein s7 from bacillus stearothermophilus.overproduction and crystallization of bacillus stearothermophilus ribosomal protein s7 (bsts7), a primary 16s rrna binding protein and also a translational repressor protein, have been performed to analyze its three-dimensional structure by x-ray crystallography. ribosomal protein bsts7 was expressed in the cytoplasmic fraction of the e. coli cells and purified to homogeneity. this recombinant bsts7 was used to produce crystals with p2(1) symmetry that diffracted to 2.5 a resolution which are su ...19979356300
cloning of genes of the aminopeptidase t family from thermus thermophilus hb8 and bacillus stearothermophilus ncib8924: apparent similarity to the leucyl aminopeptidase family.to obtain genes with sequence similarity to aminopeptidase t (ap-t) of thermus aquaticus yt-1, we cloned the genes encoding aminopeptidase th (ap-th) from thermus thermophilus hb8 and aminopeptidase ii (apii) from bacillus stearothermophilus ncib8924. the ap-th gene encoded a polypeptide of 408 amino acid residues and the deduced molecular weight of this subunit was 45,015. the apii gene encoded a polypeptide of 413 amino acid residues with a deduced molecular weight of 46,207. the extent of ami ...19979362117
cloning and characterization of the arginine-specific carbamoyl-phosphate synthetase from bacillus stearothermophilus.bacillus stearothermophilus contains two carbamoyl-phosphate synthetases (cps), one specific for pyrimidine biosynthesis and the other for arginine biosynthesis. the pyrimidine-specific cps is repressed by exogenous pyrimidines, and its activity is inhibited by ump and activated by 5-phospho-alpha-d-ribosyl diphosphate. the arginine-specific cps is similarly repressed by exogenous arginine but its activity is not sensitive to these or other potential effectors. each of the two enzymes consist of ...19979370352
entamoeba histolytica: computer-assisted modeling of phosphofructokinase for the prediction of broad-spectrum antiparasitic agents.pyrophosphate-dependent phosphofructokinase (ppi-pfk) is the rate-limiting glycolytic enzyme found in the pathogenic protists entamoeba histolytica, giardia lamblia, toxoplasma gondii, trichomonas vaginalis, and naegleria fowleri. the enzyme differs significantly from atp-dependent phosphofructokinases found in humans and as such represents an important drug target. current therapy for infections caused by these pathogens is inadequate, especially for children, pregnant women, and the immune com ...19979371084
[suitability of bacillus subtilis and bacillus stearothermophilus spores as test organism bioindicators for detecting superheating of steam].biological indicators used to test sterilisation procedures for their efficacy consist of a so-called germ carrier to which the microorganisms used as test organisms adhere. in previous papers we demonstrated that carriers made of filter paper on contact with saturated steam show superheating while carriers made of glass fibre fleece as well as wetted filter paper do not. using spores of bacillus subtilis and bacillus stearothermophilus as test organisms we have now investigated whether and to w ...19979376061
evaluation of a new device for sterilizing dental high-speed handpieces.dental high-speed turbines and handpieces can take up and expel microorganisms during operation and thus need regular sterilization. this study established a method for validating devices used to sterilize high-speed turbines and handpieces. the air and water channels and turbine chambers were contaminated with suspensions of streptococcus salivarius or endospores of bacillus stearothermophilus. the effect of flushing and/or autoclaving performed by a new device combining both procedures was eva ...19979394384
structural and mechanistic studies on chloroplast translational initiation factor 3 from euglena gracilis.chloroplast translational initiation factor 3 (if3chl) from euglena gracilis contains a central region (homology domain) that is homologous to prokaryotic if3. the homology domain is preceded by a long nh2-terminal extension (head), and followed by a 64 amino acid cooh-terminal extension (tail). sequences in these extensions reduce the activity of the homology domain. to gain insight into these effects, a possible three-dimensional structure for the homology region of if3chl has been modeled usi ...19979398204
a novel cytochrome b(o/a)3-type oxidase from bacillus stearothermophilus catalyzes cytochrome c-551 oxidation.gram-positive thermophilic bacillus species contain cytochrome caa3-type cytochrome c oxidase as their main terminal oxidase in the respiratory chain. to identify alternative oxidases, we isolated several mutants from b. stearothermophilus defective in the caa3-type oxidase activity [sakamoto, j. et al (1996) fems microbiol. lett. 143, 151-158]. a novel oxidase was isolated from membrane preparations of one of the mutants, k17. the oxidase was composed of two subunits with molecular masses of 56 ...19979399580
evaluation of selected antibiotic residue screening tests for milk from individual cows and examination of factors that affect the probability of false-positive outcomes.total composite milk samples from 131 cows in one herd were analyzed. eight beta-lactam residue screening tests were evaluated for performance using milk from individual cows and factors that affect the rate of false-positive outcomes were determined. cows were not treated with an antibiotic for at least 30 d prior to sampling. tests evaluated were delvotest p, charm cowside, charm farm, penzyme, valio t101, lactek, cite probe, and charm bacillus stearothermophilus disk assay. cows averaged 155 ...19979406098
adaptation of microorganisms and their transport systems to high temperatures.growth of bacteria and archaea has been observed at temperatures up to 95 and 110 degrees c, respectively. these thermophiles are adapted to environments of high temperature by changes in the membrane lipid composition, higher thermostabilities of the (membrane) proteins, higher turnover rates of the energy transducing enzymes, and/or the (exclusive) use of sodium-ions rather than protons as coupling ion in energy transduction. the proton permeability of the cytoplasmic membrane of bacteria and ...19979406426
sequence and structural comparison of thermophilic phosphoglycerate kinases with a mesophilic equivalent.the monomeric glycolytic enzyme phosphoglycerate kinase (pgk) has been used as a model system to study protein thermostability. the primary sequence of this enzyme has been elucidated from 47 species to date. although only 42 amino acids are totally conserved, most of which line the active site cleft, the protein is structurally conserved. this is achieved by making conservative changes to maintain the same secondary and tertiary folds. the crystal structures of 5 pgk enzymes have been solved by ...19979406428
expression, purification, and crystallization of two isozymes of 6-phosphoglucose isomerase of bacillus stearothermophilus.two isozymes of 6-phosphoglucose isomerase (phosphoglucose isomerase a & phosphoglucose isomerase b), isolated from bacillus stearothermophilus, have been overexpressed in escherichia coli strain df2145 and purified to homogeneity. crystals of both isozymes have been obtained by the vapor diffusion method. the crystals of phosphoglucose isomerase a have unit cell dimensions a = b = 132.0 a, c = 183.6 a, and diffract to about 2.8 a resolution. an analysis of the reflection data indicates that the ...19979417984
effects of a lipophilic environmental pollutant (ddt) on the phospholipid and fatty acid contents of bacillus stearothermophilus.cultures of bacillus stearothermophilus grown in a complex medium containing 1 microm ddt, exhibited longer lag adapting periods, decreased specific growth rates, and lower growth yield as compared to control cultures. the membrane lipid composition from cells grown in the presence of the insecticide was significantly different from that of control cells. the effects of ddt (2,2-bis(p-chlorophenyl)-1, 1,1-trichloroethane) on growth and lipid composition of bacterial cells were also determined in ...19979419252
[use of a microwave device for dental instrument sterilization: possibilities and limitations].the aim of the present study was the evaluation of the effectiveness as sterilizer of a modified version of a microwave device, which was previously tested by the same authors and found unsatisfactory.19979432563
extreme stabilization of a thermolysin-like protease by an engineered disulfide bond.the thermal inactivation of broad specificity proteases such as thermolysin and subtilisin is initiated by partial unfolding processes that render the enzyme susceptible to autolysis. previous studies have revealed that a surface-located region in the n-terminal domain of the thermolysin-like protease produced by bacillus stearothermophilus is crucial for thermal stability. in this region a disulfide bridge between residues 8 and 60 was designed by molecular modelling, and the corresponding sing ...19979111013
translational initiation factor if2 from bacillus stearothermophilus: a spectroscopic and microcalorimetric study of the c-domain.conformation and stability of the c-terminal domain of initiation factor if2 from bacillus stearothermophilus were analyzed by circular dichroism, fluorescence and raman spectroscopy, and microcalorimetry under different solvent conditions. from circular dichroism and raman measurements, if2c at neutral ph can be classified as an alpha + beta protein. solvent perturbation and raman spectroscopy indicate a high accessibility of the tyrosine residues in the native protein. the gdn/hcl-induced unfo ...19979115993
structure-function relationships within the peptide deformylase family. evidence for a conserved architecture of the active site involving three conserved motifs and a metal ion.thermus thermophilus peptide deformylase was characterized. its enzymatic properties as well as its organization in domains proved to share close resemblances with those of the escherichia coli enzyme despite few sequence identities. in addition to the hexxh signature sequence of the zinc metalloprotease family, a second short stretch of strictly conserved amino acids was noticed, egcls, the cysteine of which corresponds to the third zinc ligand. the study of site-directed mutants of the e. coli ...19979126850
contribution of a salt bridge to the thermostability of dna binding protein hu from bacillus stearothermophilus determined by site-directed mutagenesis.we have employed site-directed mutagenesis to evaluate the contribution of the amino acid residues, glu34, arg37, and lys38, to the thermostability of the thermophilic protein, bacillus stearothermophilus dna binding protein hu (bsthu), relative to the mesophilic homologue, bacillus subtilis hu (bsuhu). mutants bsthu-e34d and bsthu-k38n, in which glu34 and lys38 were individually replaced by the corresponding residues, asp34 and asn38, in bsuhu, exhibited decreased thermostabilities by 2.08 and ...19979133613
identification of a novel gene cluster participating in menaquinone (vitamin k2) biosynthesis. cloning and sequence determination of the 2-heptaprenyl-1,4-naphthoquinone methyltransferase gene of bacillus stearothermophilus.we recently described the isolation and sequence analysis of a dna region containing the genes of bacillus stearothermophilus heptaprenyl diphosphate synthase, which catalyzes the synthesis of the prenyl side chain of menaquinone-7 of this bacterium. sequence analyses revealed the presence of three open reading frames (orfs), designated as orf-1, orf-2, and orf-3, and the structural genes of the heptaprenyl diphosphate synthase were proved to consist of orf-1 (heps-1) and orf-3 (heps-2) (koike-t ...19979139683
immunomagnetic detection of bacillus stearothermophilus spores in food and environmental samples.there are currently no methods for the rapid and sensitive detection of bacterial spores that could be used to direct raw materials containing high spore loads away from products that pose a food safety risk. existing methods require an overnight incubation, cannot detect spores below 10(5) cfu/ml, or are not specific to particular species. this work describes a method to specifically detect < 10(4) cfu of bacterial spores per ml within 2 h. polyclonal antibodies to bacillus stearothermophilus s ...19979143097
the stability and dynamics of ribosomal protein l9: investigations of a molecular strut by amide proton exchange and circular dichroism.nuclear magnetic resonance and circular dichroism experiments were used to investigate the stability and dynamic aspects of ribosomal protein l9 from bacillus stearothermophilus in solution. this unusually shaped protein, with its two widely spaced rna-binding domains linked by a connecting helix, has been hypothesized to serve as a "molecular strut", most likely playing a role in ribosome assembly and/or maintaining the catalytically active conformation of ribosomal rna. protection factors for ...19979159485
human tyrosyl-trna synthetase shares amino acid sequence homology with a putative cytokine.to test the hypothesis that trnatyr recognition differs between bacterial and human tyrosyl-trna synthetases, we sequenced several clones identified as human tyrosyl-trna synthetase cdnas by the human genome project. we found that human tyrosyl-trna synthetase is composed of three domains: 1) an amino-terminal rossmann fold domain that is responsible for formation of the activated e.tyr-amp intermediate and is conserved among bacteria, archeae, and eukaryotes; 2) a trna anticodon recognition dom ...19979162081
crystallographic analysis of phosphoglycerate kinase from the hyperthermophilic bacterium thermotoga maritima.phosphoglycerate kinase from the hyperthermophilic bacterium thermotoga maritima has been co-crystallized with its substrate 3-phosphoglycerate and the atp analogue amp-pnp using the vapour diffusion method. crystals were obtained from a solution containing polyethylene glycol (mw 3000/8000) as precipitating agent. a complete diffraction data set from orthorhombic crystals was collected up to 2.0 a resolution. the tmpgk crystallizes in the space group p2(1)2(1)2 (cell dimensions: a = 62.0 a, b = ...19979165089
the n-terminal sequence of lactococcus lactis phosphoglucose isomerase purified by affinity chromatography differs from the other species.a specific monoclonal antibody, m3a, was produced to rapidly purify lactococcus lactis phosphoglucose isomerase (pgi) for amino acid sequence analysis. m3a recognized the lac. lactis pgi specifically and sensitively with both enzyme-linked immunosorbent assay and western blot analysis. the enzyme was rapidly purified to a specific activity of 21.8 u/mg with a yield of 20% by a three-step procedure, including m3a-bound sepharose chromatography. the specific activity of pgi was increased about 64. ...19979169021
purification and characterization of thermostable d-hydantoinase from thermophilic bacillus stearothermophilus sd-1.a thermostable d-hydantoinase of thermophilic bacillus stearothermophilus sd-1 was purified to homogeneity using an immuno-affinity chromatography. the affinity chromatography that employed polyclonal antibody immobilized on sepharose 4b was simple to operate and gave a purification yield of 60% of enzyme activity. molecular mass of the enzyme was determined to be about 133.9 kda by gel filtration chromatography and the molecular mass of the subunit was 54 kda on sds-page. mass spectrometric ana ...19979170256
a crystallographic comparison between mutated glyceraldehyde-3-phosphate dehydrogenases from bacillus stearothermophilus complexed with either nad+ or nadp+.mutations have been introduced in the cytosolic glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus in order to convert its cofactor selectivity from a specificity towards nad into a preference for nadp. in the b-s mutant, five mutations (l33t, t34g, d35g, l187a, p188s) were selected on the basis of a sequence alignment with nadp-dependent chloroplastic gapdhs. in the d32g-s mutant, two of the five mutations mentioned above (l187a, p188s) have been used in combinati ...19979175858
autosterilization of biodegradable implants by injection molding process.sterilization of degradable implants by standard procedures may damage the parts due to the labile chemical nature of the polymers. this study examined whether the injection molding process used for the production of polymeric parts may itself sterilize the implant due to high temperature, pressure, and shear forces applied. poly-d,l-lactic acid (pdlla) and poly-l-lactic acid (plla) granules were contaminated with thermoresistant spores of bacillus stearothermophilus (>10(5) spores/g). sterile a ...19979178738
comparative enzymatic properties of gapb-encoded erythrose-4-phosphate dehydrogenase of escherichia coli and phosphorylating glyceraldehyde-3-phosphate dehydrogenase.gapb-encoded protein of escherichia coli and glyceraldehyde-3-phosphate dehydrogenase (gapdh) share more than 40% amino acid identity. most of the amino acids involved in the binding of cofactor and substrates to gapdh are conserved in gapb-encoded protein. this enzyme shows an efficient non-phosphorylating erythrose-4-phosphate dehydrogenase activity (zhao, g., pease, a. j., bharani, n., and winkler, m. e. (1995) j. bacteriol. 177, 2804-2812) but a low phosphorylating glyceraldehyde-3-phosphate ...19979182530
evidence that the n-terminal part of the s-layer protein from bacillus stearothermophilus pv72/p2 recognizes a secondary cell wall polymer.the s-layer of bacillus stearothermophilus pv72/p2 shows oblique lattice symmetry and is composed of identical protein subunits with a molecular weight of 97,000. the isolated s-layer subunits could bind and recrystallize into the oblique lattice on native peptidoglycan-containing sacculi which consist of peptidoglycan of the a1gamma chemotype and a secondary cell wall polymer with an estimated molecular weight of 24,000. the secondary cell wall polymer could be completely extracted from peptido ...19979190804
species-specific trna recognition in relation to trna synthetase contact residues.in spite of variations in the sequences of trnas, the genetic code (anticodon trinucleotides) is conserved in evolution. however, non-anticodon nucleotides which are species specific are known to prevent a given trna from functioning in all organisms. conversely, species-specific trna contact residues in synthetases should also prevent cross-species acylation in a predictable way. to address this question, we investigated the relatively small tyrosine trna synthetase where contacts of escherichi ...19979192996
effectiveness of salt versus glass bead sterilizers.microorganisms can be removed from dental instruments by various methods, including treatment in salt and glass bead sterilizers. however, no rigorous, controlled, in vivo or in vitro studies have been performed to verify the respective efficiencies of these methods. the goals of this study were to determine if the positioning of instruments at the centre or edge of a salt sterilizer results in differential sterilization effectiveness, and to compare the effectiveness of salt sterilizers relativ ...19979203778
investigation of pulsed light for terminal sterilization of wfi filled blow/fill/seal polyethylene containers.a study was performed to assess the ability of pulsed light to sterilize water for injection in blow/fill/seal polyethylene containers. pulsed light uses intense, short duration flashes of broad spectrum white light to produce high levels of microbial kill. in a first phase of testing, containers of 0.5, 5, 15, and 120 ml nominal volume were inoculated with bacillus pumilus endospores, bacillus subtilus variety niger strain globigii endospores, bacillus stearothermophilus endospores, and aspergi ...19979203823
general structure/function properties of microbial methionyl-trna synthetases.alignment of the sequences of methionyl-trna synthetases from various microbial sources shows low levels of identities. however, sequence identities are clustered in a limited number of sites, most of which contain peptide patterns known to support the activity of the escherichia coli enzyme. in the present study, site-directed mutagenesis was used to probe the role of these conserved residues in the case of the bacillus stearothermophilus methionyl-trna synthetase. the b. stearothermophilus enz ...19979208948
cloning and nucleotide sequence of bacillus stearothermophilus pyruvate carboxylase.a gene for prokaryotic pyruvate carboxylase (pc) was cloned from bacillus stearothermophilus. it has an open reading frame of 3441 base pairs which can code for a protein of 128,353 da. not only the molecular size and domain organization but also the deduced amino acid sequence of b. stearothermophilus pc are similar to those of eukaryotic pcs.19979210587
sporicidal activity of chemical and physical tissue fixation methods.the effects of alcohol based fixation and microwave stimulated alcohol fixation were investigated on spores of bacillus stearothermophilus and bacillus subtilis (var. niger).19979215128
crystal structure of a thermostable bacillus dna polymerase i large fragment at 2.1 a resolution.the study of dna polymerases in the pol l family is central to the understanding of dna replication and repair. dna polymerases are used in many molecular biology techniques, including pcr, which require a thermostable polymerase. in order to learn about pol i function and the basis of thermostability, we undertook structural studies of a new thermostable dna polymerase.19979016716
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 ...19979023926
two amino acid amidohydrolase genes encoding l-stereospecific carbamoylase and aminoacylase are organized in a common operon in bacillus stearothermophilus.the l-carbamoylase gene (amab) upstream of the previously detected l-aminoacylase gene (amaa) in the bacillus stearothermophilus ncib8224 strain was identified in this study. the amab and amaa genes are cotranscribed as a single mrna from the same transcriptional start. the two-ama-gene operon is conserved in b. stearothermophilus strains. a cross-activity of l-carbamoylase towards respective substrates for l-aminoacylase supports the hypothesis of a common ancestor for both amino acid amidohydr ...19979023955
characterization of two terminal oxidases in bacillus brevis and efficiency of energy conservation of the respiratory chain.the respiratory chain of bacillus brevis was analyzed. resting cells showed an h+/o ratio of 4.8-5.2 (5.01+/-0.26), when measured using an oxygen pulse method with endogenous substrates. this value is intermediate between those of bacillus subtilis (about 4), which predominantly expresses cytochrome aa3-type quinol oxidase, and bacillus stearothermophilus (about 6), which has quinol cytochrome c reductase plus caa3-type cytochrome c oxidase. measurement of respiration with various substrates, an ...19979443812
comparative study of the toxic actions of 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane and 2,2-bis(p-chlorophenyl)-1,1-dichloroethylene on the growth and respiratory activity of a microorganism used as a model.a strain of bacillus stearothermophilus was used as a model for a comparative study of the toxic effect of 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane and 2,2-bis(p-chlorophenyl)-1,1-dichloroethylene. bacterial growth, the o2 consumption rate, and respiration-related enzymatic activities provided quantitative data in agreement with results reported for other systems. the use of this bacterium for screening for chemical toxicity is discussed.19979471966
dose-response effects of raw potato starch on small-intestinal escape, large-bowel fermentation and gut transit time in the rat.this study was designed to quantify starch digestion within the small and large bowels separately when raw potato starch (rps) was included at 0-240 g/kg in diets fed to growing male wistar rats. rps was incorporated in the diets at the expense of maize starch which was expected to be almost completely digested in the small bowel. the digestibility of the maize starch was 0.99 but only 0.28 of the rps was digested before the terminal ileum so that with increasing intakes of rps there was a progr ...19979497449
sterilization of slide sheath anesthetic injection systems placed within sharps containers.the purpose of this study was to evaluate the effect that two steam autoclaves and an unsaturated chemical vapor sterilizer had on killing bacterial endospores present on commercial spore strips or applied to sterile anesthetic injection systems placed within sharps containers. three types of sterilizers were used: a gravity steam autoclave, a high vacuum steam autoclave and an unsaturated chemical vapor sterilizer. the microbial challenge for the sterilizers were bacillus stearothermophilus spo ...19979517339
cleaning of endodontic files, part i: the effect of bioburden on the sterilization of endodontic files.ninety-two new endodontic files were randomly assigned to five groups with varying parameters of contamination, cleaning method, and sterilization (steam or chemical). files were instrumented in bovine teeth to accumulate debris and a known contaminant, bacillus stearothermophilus. positive controls produced growth on both t-soy agar plates and in t-soy broth. negative controls and experimental files (some with heavy debris) failed to produce growth. the results showed that there was no signific ...19979594742
chemical modifications of bacillus subtilis tryptophanyl-trna synthetase.a concerted conformational change in bacillus subtilis tryptophanyl-trna synthetase (trprs) was evident from previous fluorescence on the quenching of the single trp residue trp-92 in the 4ftrp-amp complexed enzyme. in this study, chemical modifications of the b. subtilis trprs were employed to further characterize this conformational change, with the single trp residue serving as a marker for monitoring the change. modifications of the enzyme by means of the trp-specific agent n-bromosuccinimid ...19979599659
use of 3-d computer modelling and kinetic studies to analyse grapefruit pyrophosphate-dependent phosphofructokinase.the glycolytic reaction of grapefruit ppi-dependent phosphofructokinase (pfp) depends on the presence of fru-2,6-p2 (ka = 6.7 nm). this molecule was further demonstrated in grapefruit juice sac cells. citrate, alpha-ketoglutarate and isocitrate competitively inhibited the binding of fru-2,6-p2 to pfp. the affinity for fru-6-p (km = 159 microm) and ppi (km = 33 microm) were not affected by the addition of these molecules. in the gluconeogenic reaction, the presence of fru-2,6-p2 did not affect th ...19979493054
improving the thermostability of bacillus stearothermophilus neutral protease by introducing proline into the active site helix.a proline residue was introduced into the n-terminus (ile140 and asp141), the middle (leu147) and the c-terminus (asp153) of the active site helix of bacillus stearothermophilus neutral protease for comparing the effects on the thermostability. introduction of a proline residue into the n-terminus at sites 140 and 141 increased the half-survival temperature (hst) by 7.5 and 2.8 degrees c, respectively, from 68.3 degrees c of the wild-type enzyme. a proline residue at leu147 decreased the hst by ...19979514114
purification and properties of mangano-superoxide dismutase from a strain of alkaliphilic bacillus.a mangano-superoxide dismutase (ec 1.15.1.1) was purified to homogeneity from a strain of alkaliphilic bacillus for the first time. the purified protein, with an isoelectric point of ph 4.5, had a molecular mass of approximately 50 kda and consisted of two identical subunits (25 kda). the n-terminal amino acid sequence was ala-tyr-lys-leu-pro-glu-leu-pro-tyr-ala-ala-asn-ala-leu-glu-pro-his-ile- asp-glu-ala. the optimum ph and temperature for the reaction were 7.5 and 35 degrees c, respectively. ...19979680305
[screening of thermophilic superoxide dismatase producing strain and some properties of the enzyme].a thermophilic sod producing strain was obtained from the bacteria preserved in our lab. its content of sod was 8774u/g fresh cells. the strain can tolerate 0.4% h2o2 and 70 degrees c, and it has outstanding culture characteristics. it was identified as bacillus stearothermophilus, called b.s 211-15. crude sod was extracted from b.s 211-15, the recovery of total activity was 69.5%, the specific activity was 1793u/mg protein. the enzyme showed fine heat stability, ph stability and good proteinase ...19979863206
enzymatic assay of d-mannose in serum.we describe a new and improved enzymatic assay for determining the concentration of d-mannose in sera. serum d-glucose is selectively converted to glucose-6 phosphate with the highly specific thermostable glucokinase (ec 2.7.1.2) from bacillus stearothermophilus. the anionic reaction products and excess substrates are removed by a rapid and simple anion-exchange chromatography step in microcentrifuge spin columns. d-mannose in the glucose-depleted sample is then assayed spectrophotometrically by ...19979068599
translation initiation factor if2 of the myxobacterium stigmatella aurantiaca: presence of a single species with an unusual n-terminal sequence.the structural gene for translation initiation factor if2 (infb) was isolated from the myxobacterium stigmatella aurantiaca on a 5.18-kb bamhi genomic restriction fragment. the infb gene (ca. 3.16 kb) encodes a 1,054-residue polypeptide with extensive homology within its g domain and c terminus with the equivalent regions of if2s from escherichia coli, bacillus subtilis, bacillus stearothermophilus, and streptococcus faecium. the n-terminal region does not display any significant homology to oth ...19979079922
identification of the serine acetyltransferase gene of staphylococcus xylosus.a transposon tn917-induced mutant strain of staphylococcus xylosus was isolated that required exogenous cysteine for growth. the transposon was found to reside within a gene, designated cyse, encoding a protein of 216 amino acids with a high level of similarity to bacterial serine acetyltransferases. the cyse::tn917 mutant completely lost serine acetyltransferase activity, which is easily detectable in the wild-type strain. in addition, the mutant strain could no longer grow in minimal medium wi ...19979084146
engineering thermolysin-like proteases whose stability is largely independent of calcium.thermal stability of the thermolysin-like protease produced by bacillus stearothermophilus (tlp-ste) is highly dependent on calcium at concentrations in the millimolar range. we describe the rational design and production of a fully active tlp-ste variant whose stability is only slightly dependent on calcium concentration.19979089298
lysyl-trna synthetase from bacillus stearothermophilus. formation and isolation of an enzyme-lysyladenylate complex and its analogue.the formation of an enzyme.lysyladenylate complex was studied with a highly purified lysyl-trna synthetase [l-lysine:trnalys ligase (amp-forming); ec 6.1.1.6] from bacillus stearothermophilus. the apparent dissociation equilibrium constants of the enzyme with l-lysine and atp in the process of the complex formation were estimated to be 50.9 and 15.5 microm, respectively, at ph 8.0, 30 degrees c, by fluorometric measurement. the isolated enzyme.lysyladenylate complex was relatively stable with a ...19979089397
structure and regulation of expression of the bacillus subtilis valyl-trna synthetase gene.we have sequenced the valyl-trna synthetase gene (vals) of bacillus subtilis and found an open reading frame coding for a protein of 880 amino acids with a molar mass of 101,749. the predicted amino acid sequence shares strong similarity with the valyl-trna synthetases from bacillus stearothermophilus, lactobacillus casei, and escherichia coli. extracts of b. subtilis strains overexpressing the vals gene on a plasmid have increased valyl-trna aminoacylation activity. northern analysis shows that ...19979098041
overexpression of bsobi restriction endonuclease in e. coli, purification of the recombinant bsobi, and identification of catalytic residues of bsobi by random mutagenesis.bsobi is a type ii restriction enzyme found in bacillus stearothermophilus jn209 that recognizes the symmetric sequence 5'-cycgrg-3' (y=c or t; r=a or g) and cleaves between the first and second base to generate a four-base 5' extension. the cloning and sequencing of bsobi restriction-modification system has been described by ruan et al. [mol. gen. genet. 252 (1996) 695-699]. here we report the overexpression of bsobi restriction endonuclease gene in e. coli by insertion of the endonuclease gene ...19979099856
primary structure and strand specificity of bstf5i-1 dna methyltransferase which recognizes 5'-ggatg-3'.the gene for bstf5i-1 dna-methyltransferase (mtase) from bacillus stearothermophilus f5 (a foki isoschizomer, recognizing 5'-ggatg-3') was cloned and its nucleotide (nt) sequence was determined. analysis of deduced amino acid (aa) sequence shows that m x bstf5i-1 belongs to d21 class of mtases and has a little homology with m x foki. m x bstf5i-1 modifies only the upper strand of recognition sequence (5'-ggatg-3').19979099883
conversion from archaeal geranylgeranyl diphosphate synthase to farnesyl diphosphate synthase. two amino acids before the first aspartate-rich motif solely determine eukaryotic farnesyl diphosphate synthase activity.farnesyl diphosphate (fpp) and geranylgeranyl diphosphate (ggpp) are precursors for a variety of important natural products, such as sterols, carotenoids, and prenyl quinones. although fpp synthase and ggpp synthase catalyze similar consecutive condensations of isopentenyl diphosphate with allylic diphosphates and have several homologous regions in their amino acid sequences, nothing is known about how these enzymes form the specific products. to locate the region that causes the difference of f ...19979030588
the utilization of rapd-pcr for identifying thermophilic and mesophilic bacillus species.a random amplified polymorphic dna fingerprinting assay has been optimized that is able to discriminate between numerous thermophilic and mesophilic bacillus species and strains. included in the analyses are thermophilic (able to grow at 55 degrees c) strains of bacillus stearothermophilus, b. kaustophilus, b. coagulans, b. sphaericus, b. thermodenitrificans, b. thermocatenulatus, b. thermoleovorans, b. licheniformis, b. brevis, b. thermoglucosidasius, b. caldolyticus, b. caldotenax, b. caldovel ...19979037767
molecular characterization of the bacillus stearothermophilus pv72 s-layer gene sbsb induced by oxidative stress.s-layer protein variation from a hexagonally ordered (sbsa; 130 kda) to a obliquely ordered (sbsb; 98 kda) protein in bacillus stearothermophilus pv72 is mediated by an increased oxygen supply. to elucidate the molecular basis of s-layer protein variation in b. stearothermophilus pv72, the sbsb gene, coding for the 98-kda protein, was cloned by means of inverse pcr technology and sequenced. the sbsb coding region cloned in puc18 was expressed in escherichia coli, without its own regulatory upstr ...19979045827
protein engineering tests of a homology model of plasmodium falciparum lactate dehydrogenase.this paper describes the testing of a homology model of plasmodium falciparum lactate dehydrogenase (pfldh) by protein engineering. the model had been validated in structural terms. it suggests explanations of the unusual properties of pfldh (compared with all other ldhs). these unusual features are a lack of substrate inhibition, high activity with the synthetic coenzyme 3-acetylpyridine adenine dinucleotide (apad+) and changes in residues at previously conserved positions. pfldh shows several ...19979051732
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