| novel bifunctional hyperthermostable carboxypeptidase/aminoacylase from pyrococcus horikoshii ot3. | genome sequencing of the thermophilic archaeon pyrococcus horikoshii ot3 revealed a gene which had high sequence similarity to the gene encoding the carboxypeptidase of sulfolobus solfataricus and also to that encoding the aminoacylase from bacillus stearothermophilus. the gene from p. horikoshii comprises an open reading frame of 1,164 bp with an atg initiation codon and a tga termination codon, encoding a 43,058-da protein of 387 amino acid residues. however, some of the proposed active-site r ... | 2001 | 11157230 |
| s-layer variation in bacillus stearothermophilus pv72 is based on dna rearrangements between the chromosome and the naturally occurring megaplasmids. | bacillus stearothermophilus pv72 expresses different s-layer genes (sbsa and sbsb) under different growth conditions. no stretches of significant sequence identity between sbsa and sbsb were detected. in order to investigate s-layer gene regulation in b. stearothermophilus pv72, we characterized the upstream regulatory region of sbsa and sbsb by sequencing and primer extension analysis. both genes are transcribed from unique but different promoters, independently of the growth phase. localizatio ... | 2001 | 11160098 |
| characterization of bsemii, a new type iv restriction-modification system, which recognizes the pentanucleotide sequence 5'-ctcag(n)(10/8)/. | we report the properties of the new bsemii restriction and modification enzymes from bacillus stearothermophilus isl 15-111, which recognize the 5'-ctcag sequence, and the nucleotide sequence of the genes encoding them. the restriction endonuclease r.bsemii makes a staggered cut at the tenth base pair downstream of the recognition sequence on the upper strand, producing a two base 3'-protruding end. magnesium ions and s:-adenosyl-l-methionine (adomet) are required for cleavage. s:-adenosylhomocy ... | 2001 | 11160921 |
| fragmentary form of thermostable leucine dehydrogenase of bacillus stearothermophilus: its construction and reconstitution of active fragmentary enzyme. | x-ray crystallographic studies revealed that various amino acid dehydrogenases fold into two domains in each subunit, a substrate-binding domain and an nad(p)(+)-binding domain (baker, p. j., turnbull, a. p., sedelnikova, s. e., stillman, t. j., and rice, d. w. (1995) structure 3, 693-705). to elucidate the function and folding process of these two domains, we have genetically constructed a fragmentary form of thermostable leucine dehydrogenase of bacillus stearothermophilus consisting of an n-t ... | 2001 | 11162651 |
| the growth of bacillus stearothermophilus on stainless steel. | to determine the potential for bacillus stearothermophilus cells to form biofilms of significance in dairy manufacture. | 2001 | 11168716 |
| thermal sterilization of heat-sensitive products using high-temperature short-time sterilization. | high-temperature short-time (htst) sterilization with a continuous-flow sterilizer, developed for this study, was evaluated. the evaluation was performed with respect to (a) the chemical degradation of two heat-sensitive drugs in htst range (140-160 degrees c) and (b) the microbiological effect of htst sterilization. degradation kinetics of two heat-sensitive drugs showed that a high peak temperature sterilization process resulted in less chemical degradation for the same microbiological effect ... | 2001 | 11170021 |
| protein-protein interaction between bacillus stearothermophilus tyrosyl-trna synthetase subdomains revealed by a bacterial two-hybrid system. | we have recently developed a bacterial two-hybrid system (bacth), based on functional complementation between two fragments of the catalytic domain of bordetella pertussis adenylate cyclase (ac), that allows an easy in vivo screening and selection of functional interactions between two proteins in escherichia coli. in this work, we have further explored the potentialities of the bacth system to study protein-protein interactions, using as a model, the interactions between various subdomains of t ... | 2001 | 11200232 |
| effect of ph on the heat resistance of spores comparison of two models. | all published models describing the effect of ph on the heat resistance of spores can be regarded either as a linear first degree equation or a linear second degree equation. this work aimed to compare both models from three sets of published data for, clostridium sporogenes and bacillus stearothermophilus, respectively. the relative quality of fit of each model with respect to the other depends on the species, the strain and the heating temperature. parameter estimation was more reliable for th ... | 2001 | 11205953 |
| gene cloning and characterization of alpha-glucuronidase of bacillus stearothermophilus no. 236. | the alpha-glucuronidase gene of bacillus stearothermophilus no. 236 was cloned, sequenced, and expressed in escherichia coli. the gene, designated agua, encoded a 691-residue polypeptide with calculated molecular weight of 78,156 and pi of 5.34. the alpha-glucuronidase produced by a recombinant e. coli strain containing the agua gene was purified to apparent homogeneity and characterized. the molecular weight of the alpha-glucuronidase was 77,000 by sds-page and 161,000 by gel filtration; the fu ... | 2000 | 11210113 |
| crystal structure of bacillus stearothermophilus alpha-amylase: possible factors determining the thermostability. | the crystal structure of a thermostable alpha-amylase from bacillus stearothermophilus (bsta) has been determined at 2.0 a resolution. the main-chain fold is almost identical to that of the known crystal structure of bacillus licheniformis alpha-amylase (bla). bla is known to be more stable than bsta. a structural comparison between the crystal structures of bsta and bla showed significant differences that may account for the difference in their thermostabilities, as follows. (i) the two-residue ... | 2001 | 11226887 |
| enzyme-induced covalent modification of methionyl-trna synthetase from bacillus stearothermophilus by methionyl-adenylate: identification of the labeled amino acid residues by matrix-assisted laser desorption-ionization mass spectrometry. | methionyl-trna synthetase (metrs) from bacillus stearothermophilus was shown to undergo covalent methionylation by a donor methionyl-adenylate, the mixed carboxylic-phosphoric acid anhydride synthesized by the enzyme itself. covalent reaction of methionyl-adenylate with the synthetase or other proteins proceeds through the formation of an isopeptide bond between the carboxylate of the amino acid and the epsilon-nh2 group of lysyl residues. the stoichiometries of labeling, as followed by tca prec ... | 2000 | 11233169 |
| rapid detection of bacillus stearothermophilus using impedance-splitting. | an impedance splitting method was used to detect bacillus stearothermophilus in suspension and attached to stainless steel surfaces. the effects of bacterial metabolism on the impedance of the culture medium and the ionic layers of the measuring electrodes were recorded using the bactrac 4000 microorganism growth analyser. impedance changes were measured at 55 degrees c. seven of the eight media produced changes in the electrode impedance (e-value) and all media produced negligible changes in th ... | 2001 | 11240042 |
| conserved amino acids near the carboxy terminus of bacterial tyrosyl-trna synthetase are involved in trna and tyr-amp binding. | bacterial tyrosyl-trna synthetases occur in two large subfamilies, tyrrs and tyrrz, that possess about 25% amino acid identity. their amino-terminal region, the active site domain, is more conserved (>36% identity). the carboxy-terminal segment of these enzymes includes the trna binding domain and contains only few conserved residues. replacement of three of these residues in acidithiobacillus ferrooxidans tyrrz revealed that s356 and k395 play roles in trna binding, while h306, a residue at the ... | 2001 | 11240138 |
| function of the neurospora crassa mitochondrial tyrosyl-trna synthetase in rna splicing. role of the idiosyncratic n-terminal extension and different modes of interaction with different group i introns. | the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) promotes the splicing of group i introns by helping the intron rna fold into the catalytically active structure. the regions required for splicing include an idiosyncratic n-terminal extension, the nucleotide-binding fold domain, and the c-terminal rna-binding domain. here, we show that the idiosyncratic n-terminal region is in fact comprised of two functionally distinct parts: an upstream region consisting predominantl ... | 2001 | 11243805 |
| sequence analysis of the aminoacylase-1 family. a new proposed signature for metalloexopeptidases. | the amino acid sequence analysis of the human and porcine aminoacylases-1, the carboxypeptidase s precursor from saccharomyces cerevisiae, the succinyl-diaminopimelate desuccinylase from escherichia coli, haemophilus influenzae and corynebacterium glutamicum, the acetylornithine deacetylase from escherichia coli and dictyostelium discoideum and the carboxypeptidase g(2) precursor from pseudomonas strain, using the basic local alignment search tool (blast) and the position-specific iterated blast ... | 2001 | 11250542 |
| improved agar diffusion method for detecting residual antimicrobial agents. | the improved agar diffusion method for determination of residual antimicrobial agents was investigated, and the sensitivities of various combinations of test organisms and assay media were determined using 7 organisms, 5 media, and 31 antimicrobial agents. bacillus stearothermophilus and synthetic assay medium (sam) showed the greatest sensitivity for screening penicillins (penicillin g and ampicillin). the combination of bacillus subtilis and minimum medium (mm) was the most sensitive for tetra ... | 2001 | 11252480 |
| equilibrium binding studies of a tryptophan-shifted mutant of phosphofructokinase from bacillus stearothermophilus. | a tryptophan-shifted mutant of phosphofructokinase (pfk) from bacillus stearothermophilus has been constructed. this mutant, which is functionally similar to wild-type, provides the opportunity to examine the allosteric properties of pfk under equilibrium conditions. the unique fluorescence properties of the tryptophan-shifted mutant enzyme, w179f/f230w, have been utilized to deduce the thermodynamics of ligand binding and the allosteric perturbations in the absence of catalytic turnover. specif ... | 2001 | 11258913 |
| mass spectrometry as a novel approach to probe cooperativity in multimeric enzymatic systems. | investigating cooperativity in multimeric enzymes is of utmost interest to improve our understanding of the mechanism of enzymatic regulation. in the present article, we propose a novel approach based on mass spectrometry to probe cooperativity in the binding of a ligand to a multisubunit enzyme. this approach presents the selective advantage of giving a direct insight into all the subsequent ligation states that are formed in solution as the ligand is added to the enzyme. a quantitative interpr ... | 2001 | 11262155 |
| purification and characterization of a thermostable alpha-amylase from bacillus stearothermophilus. | a soil isolate of bacillus stearothermophilus was found to synthesize thermostable alpha-amylase. the enzyme was purified to homogeneity by ammonium sulfate fractionation and iecc on deae-cellulose column. the purified enzyme was considered to be a monomeric protein with a molar mass of 64 kda, as determined by sds-page. the enzyme showed a wide range of ph tolerance and maximum activity at ph 7.0. the temperature tolerance was up to 100 degrees c with more than 90% catalytic activity; the maxim ... | 2000 | 11271801 |
| comparative study of the cyclization reactions of three bacterial cyclomaltodextrin glucanotransferases. | the actions of cyclomaltodextrin glucanotransferases (cgtase; ec 2.4.1.19) from alkalophilic bacillus sp. strain a2-5a (a2-5a cgtase), bacillus macerans (bmac cgtase), and bacillus stearothermophilus (bste cgtase) on amylose were investigated. all three enzymes produced large cyclic alpha-1,4-glucans (cycloamyloses) at the early stage of the reaction, but these were subsequently converted into smaller cycloamyloses. however, the rates of this conversion differed among the three enzymes. the prod ... | 2001 | 11282590 |
| inhibition of beta-glycosidases by acarbose analogues containing cellobiose and lactose structures. | acarbose analogues, containing cellobiose and lactose structures, were prepared by reaction of the two disaccharides with acarbose and bacillus stearothermophilus maltogenic amylase. the kinetics for the inhibition by the two analogues was studied for beta-glucosidase, beta-galactosidase, cyclomaltodextrin glucanosyltransferase (cgtase), and alpha-glucosidase. both analogues were potent competitive inhibitors for beta-glucosidase, with k(i) values in the range of 0.04-2.44 microm, and the lactos ... | 2001 | 11284501 |
| direct evidence for the size and conformational variability of the pyruvate dehydrogenase complex revealed by three-dimensional electron microscopy. the "breathing" core and its functional relationship to protein dynamics. | structural studies by three-dimensional electron microscopy of the saccharomyces cerevisiae truncated dihydrolipoamide acetyltransferase (te(2)) component of the pyruvate dehydrogenase complex reveal an extraordinary example of protein dynamics. the te(2) forms a 60-subunit core with the morphology of a pentagonal dodecahedron and consists of 20 cone-shaped trimers interconnected by 30 bridges. frozen-hydrated and stained molecules of te(2) in the same field vary in size approximately 20%. analy ... | 2001 | 11285267 |
| a novel pathway of aerobic benzoate catabolism in the bacteria azoarcus evansii and bacillus stearothermophilus. | the aerobic catabolism of benzoate was studied in the gram-negative proteobacterium azoarcus evansii and in the gram-positive bacterium bacillus stearothermophilus. in contrast to earlier proposals, benzoate was not converted into hydroxybenzoate or gentisate. rather, benzoyl-coa was a product of benzoate catabolism in both microbial species under aerobic conditions in vivo. benzoyl-coa was converted into various coa thioesters by cell extracts of both species in oxygen- and nadph-dependent reac ... | 2001 | 11306574 |
| an invariant threonine is involved in self-catalyzed cleavage of the precursor protein for ornithine acetyltransferase. | in bacillus stearothermophilus ornithine acetyltransferase is a bifunctional enzyme, catalyzing the first and the fifth steps of arginine biosynthesis; it follows a ping-pong kinetic mechanism. a single chain precursor protein is cleaved between the alanine and threonine residues in a highly conserved atml sequence leading to the formation of alpha and beta subunits that assemble into a heterotetrameric 2alpha2beta molecule. the beta subunit has been shown to form an acetylated intermediate in t ... | 2001 | 11320085 |
| analysis of the structure-function relationship of the s-layer protein sbsc of bacillus stearothermophilus atcc 12980 by producing truncated forms. | the mature surface layer (s-layer) protein sbsc of bacillus stearothermophilus atcc 12980 comprises amino acids 31-1099 and self-assembles into an oblique lattice type which functions as an adhesion site for a cell-associated high-molecular-mass exoamylase. to elucidate the structure-function relationship of distinct segments of sbsc, three n- and seven c-terminal truncations were produced in a heterologous expression system, isolated, purified and their properties compared with those of the rec ... | 2001 | 11320138 |
| taxonomic study of aerobic thermophilic bacilli: descriptions of geobacillus subterraneus gen. nov., sp. nov. and geobacillus uzenensis sp. nov. from petroleum reservoirs and transfer of bacillus stearothermophilus, bacillus thermocatenulatus, bacillus thermoleovorans, bacillus kaustophilus, bacillus thermodenitrificans to geobacillus as the new combinations g. stearothermophilus, g. th. | five hydrocarbon-oxidizing strains were isolated from formation waters of oilfields in russia, kazakhstan and china. these strains were moderately thermophilic, neutrophilic, motile, spore-forming rods, aerobic or facultatively anaerobic. the g+c content of their dna ranged from 49.7 to 52.3 mol%. the major isoprenoid quinone was menaquinone-7; cellular fatty acid profiles consisted of significant amounts of iso-15:0, iso-16:0 and iso-17:0 fatty acids (61.7-86.8% of the total). based on data fro ... | 2001 | 11321089 |
| stereochemistry of family 52 glycosyl hydrolases: a beta-xylosidase from bacillus stearothermophilus t-6 is a retaining enzyme. | a beta-xylosidase from bacillus stearothermophilus t-6 assigned to the uncharacterized glycosyl hydrolase family 52 was cloned, overexpressed in escherichia coli and purified. the enzyme showed maximum activity at 65 degrees c and ph 5.6-6.3. the stereochemistry of the hydrolysis of p-nitrophenyl beta-d-xylopyranoside was followed by 1h-nuclear magnetic resonance. time dependent spectrum analysis showed that the configuration of the anomeric carbon was retained, indicating that a retaining mecha ... | 2001 | 11322943 |
| glutamic acid 160 is the acid-base catalyst of beta-xylosidase from bacillus stearothermophilus t-6: a family 39 glycoside hydrolase. | a beta-xylosidase from bacillus stearothermophilus t-6 was cloned, overexpressed in escherichia coli and purified to homogeneity. based on sequence alignment, the enzyme belongs to family 39 glycoside hydrolases, which itself forms part of the wider gh-a clan. the conserved glu160 was proposed as the acid-base catalyst. an e160a mutant was constructed and subjected to steady state and pre-steady state kinetic analysis together with azide rescue and ph activity profiles. the observed results supp ... | 2001 | 11322958 |
| cloning and functional characterization of an nad(+)-dependent dna ligase from staphylococcus aureus. | a staphylococcus aureus mutant conditionally defective in dna ligase was identified by isolation of complementing plasmid clones that encode the s. aureus liga gene. orthologues of the putative s. aureus nad(+)-dependent dna ligase could be identified in the genomes of bacillus stearothermophilus and other gram-positive bacteria and confirmed the presence of four conserved amino acid motifs, including motif i, kxdg with lysine 112, which is believed to be the proposed site of adenylation. dna se ... | 2001 | 11325928 |
| pressure denaturation of phosphorylating glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | the effects of hydrostatic pressure on apo wild-type glyceraldehyde-3-phosphate dehydrogenase (wtgapdh) from bacillus stearothermophilus (b. stearothermophilus) have been studied by fluorescence spectroscopy under pressure from 0.1 to 650 mpa. unlike yeast gapdh [ruan, k. c., and weber, g. (1989) biochemistry 28, 2144-2153], denaturation of the tetrameric apo wtgapdh from b. stearothermophilus is likely to precede dissociation into subunits. as expected, denaturation is accompanied by the loss o ... | 2001 | 11327706 |
| thermally induced changes of lipoate acetyltransferase inner core isolated from the bacillus stearothermophilus pyruvate dehydrogenase complex. | incubation at 70 degrees c converted the bacillus stearothermophilus lipoate acetyltransferase inner core into an unidentified active molecular form, x, yielding an inactive aggregate. the core and x showed similar thermostabilities, but they were different in the recovery of enzyme activity after incubation with 1.2-2.0 m guanidine hydrochloride and its subsequent removal; the core was hardly recovered, but x was well recovered. | 2001 | 11330694 |
| polymer versus monomer as displacer in immobilized metal affinity chromatography. | successful immobilized metal affinity chromatography (imac) of proteins on cu2+-iminodiacetic acid sepharose has been carried out in a displacement mode using a synthetic copolymer of vinyl imidazole and vinyl caprolactam [poly(vi-vcl)] as a displacer. vinyl caprolactam renders the co-polymer with the thermosensitivity, e.g., property of the co-polymer to precipitate nearly quantitatively from aqueous solution on increase of the temperature to 48 degrees c. a thermostable lactate dehydrogenase f ... | 2001 | 11334341 |
| spectroelectrochemical study of cellobiose dehydrogenase and diaphorase in a thiol-modified gold capillary in the absence of mediators. | a spectroelectrochemical cell was constructed from a gold capillary with 200 microm inner diameter as a working electrode. this allowed spectroelectrochemical study of liquid samples with available volumes less than 5 microl. the optical measurements were accomplished with an optical fibre spectrometer. the optical path of the cell was about 1 cm. to facilitate electrochemistry of biomolecules, the surface of the gold capillary was modified with thiols. the formal potential, e degrees', of the h ... | 2001 | 11339313 |
| ribosomal protein l5 has a highly twisted concave surface and flexible arms responsible for rrna binding. | ribosomal protein l5 is a 5s rrna binding protein in the large subunit and plays an essential role in the promotion of a particular conformation of 5s rrna. the crystal structure of the ribosomal protein l5 from bacillus stearothermophilus has been determined at 1.8 a resolution. the molecule consists of a five-stranded antiparallel beta-sheet and four alpha-helices, which fold in a way that is topologically similar to the ribonucleoprotein (rnp) domain. the molecular shape and electrostatic rep ... | 2001 | 11350033 |
| structural modeling and characterization of a thermostable lipase from bacillus stearothermophilus p1. | the moderate thermophilic bacterium bacillus stearothermophilus p1 expresses a thermostable lipase that was active and stable at the high temperature. based on secondary structure predictions and secondary structure-driven multiple sequence alignment with the homologous lipases of known three-dimensional (3-d) structure, we constructed the 3-d structure model of this enzyme and the model reveals the topological organization of the fold, corroborating our predictions. we hypothesized for this enz ... | 2001 | 11350065 |
| biochemical characterization and identification of catalytic residues in alpha-glucuronidase from bacillus stearothermophilus t-6. | alpha-d-glucuronidases cleave the alpha-1,2-glycosidic bond of the 4-o-methyl-d-glucuronic acid side chain of xylan, as a part of an array of xylan hydrolyzing enzymes. the alpha-d-glucuronidase from bacillus stearothermophilus t-6 was overexpressed in escherichia coli using the t7 polymerase expression system. the purification procedure included two steps, heat treatment and gel filtration chromatography, and provided over 0.3 g of pure enzyme from 1 l of overnight culture. based on gel filtrat ... | 2001 | 11358519 |
| method for real-time detection of inorganic pyrophosphatase activity. | a sensitive and simple method for real-time detection of inorganic pyrophosphatase (ppase) (ec 3.6.1.1) activity has been developed. the method is based on ppase-induced activation of the firefly luciferase activity in the presence of inorganic pyrophosphate (ppi). ppi inhibits the luciferase activity, but in the presence of ppase the luciferase activity is restored and the luminescence output increases. the assay yields linear responses between 8 and 500 mu. the detection limit was found to be ... | 2001 | 11373080 |
| crystallization and preliminary x-ray diffraction analysis of nadp-dependent glyceraldehyde-3-phosphate dehydrogenase of synechococcus pcc 7942. | the nadp-dependent glyceraldehyde-3-phosphate dehydrogenase (gapdh) of synechococcus pcc 7942 was crystallized in two different forms by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant. form i crystals were hexagonal, space group p6(5) or p6(1), with unit-cell parameters a = b = 91.1, c = 428.6 a, gamma = 120 degrees. form ii crystals were monoclinic, space group c2, with unit-cell parameters a = 152.3, b = 80.9, c = 213.6 a, beta = 103.1 degrees. native data wer ... | 2001 | 11375515 |
| reusable biopsy forceps: a prospective evaluation of cleaning, function, adequacy of tissue specimen, and durability. | recent studies have indicated that reusable biopsy forceps remain contaminated after reprocessing and can only be used a mean of 12 to 25 times without malfunction. because this contradicts traditional endoscopic practice, our study investigated the ability to sterilize a type of commercially available biopsy forceps and prospectively evaluated their function in vivo until malfunction and/or breakage. | 2001 | 11375582 |
| structure and expression of an amylopullulanase gene from bacillus stearothermophilus ts-23. | an amylopullulanase gene (aputs) from bacillus stearothermophilus ts-23 was cloned and characterized. aputs consisted of an open reading frame of 6054 bp encoding a protein of 2018 amino acids with a calculated m(r) of 223811. the deduced amino acid sequence revealed four highly conserved regions that are common among amylolytic enzymes. in the c-terminal region, a six-amino-acid sequence (pro-gly-ser-gly-thr-thr) is repeated nine times. it shared the highest degree of homology with the amylopul ... | 2001 | 11389673 |
| in vitro study and in vivo application of a reusable double-channel sphincterotome. | therapeutic endoscopic retrograde cholangiopancreatography (ercp) has been deemed to be a "cost-prohibitive" procedure, based upon the cumulative costs of one-time-use accessories and current reimbursement plans. one-time-use sphincterotomes comprise a significant component of that cost and, accordingly, we evaluated the disability and clinical usefulness of a recently introduced reusable double-channel sphincterotome. | 2001 | 11396756 |
| recognition of 16s rrna by ribosomal protein s4 from bacillus stearothermophilus. | protein s4 is essential for bacterial small ribosomal subunit assembly and recognizes the 5' domain (approximately 500 nt) of small subunit rrna. this study characterizes the thermodynamics of forming the s4-5' domain rrna complex from a thermophile, bacillus stearothermophilus, and points out unexpected differences from the homologous escherichia coli complex. upon incubation of the protein and rna at temperatures between 35 and 50 degrees c under ribosome reconstitution conditions [350 mm kcl, ... | 2001 | 11401563 |
| an essential residue in the flexible peptide linking the two idiosynchratic domains of bacterial tyrosyl-trna synthetases. | tyrosyl-trna synthetase (tyrrs) from bacillus stearothermophilus comprises three sequential domains: an n-terminal catalytic domain, an alpha-helical domain with unknown function, and a c-terminal trna binding domain (residues 320-419). the properties of the polypeptide segment that links the alpha-helical and c-terminal domains, were analyzed by measuring the effects of sequence changes on the aminoacylation of trna(tyr) with tyrosine. mutations f323a (phe323 into ala), s324a, and g325a showed ... | 2001 | 11401566 |
| the a245k mutation exposes another stage of the bacterial l-lactate dehydrogenase reaction mechanism. | the a245k mutant of bacillus stearothermophilus l-lactate dehydrogenase has been expressed in escherichia coli and purified. a qualitative change in the reaction mechanism prior to the hydride transfer step in the reverse direction in the mutant is revealed. both transient and steady state characteristics of the mutant are presented and show in contrast to the wild-type enzyme where a rearrangement of an enzyme-nadh-pyruvate complex is rate-limiting that in the mutant the rearrangement is much f ... | 2001 | 11401572 |
| functions of the conserved anionic amino acids and those interacting with the substrate phosphate group of phosphoglucose isomerase. | phosphoglucose isomerase catalyzes the isomerization between glucose 6-phosphate and fructose 6-phosphate in cytoplasm, and functions as autocrine motility factor and neuroleukin outside the cells. a phosphoglucose isomerase from bacillus stearothermophilus (pgia) was subjected to mutagenesis study to address the catalytic function of the conserved anionic residues and those probably interacting with the phosphate group of substrates. the results suggest that glu290 works concertedly with his311 ... | 2001 | 11418102 |
| differentiation between conformational and autoproteolytic stability of the neutral protease from bacillus stearothermophilus containing an engineered disulfide bond. | the introduction of a disulfide bond into the neutral protease from bacillus stearothermophilus by the double mutation g8c/n60c had resulted in an extremely thermostable enzyme with a half-life of 35.9 min at 92.5 degrees c [mansfeld, j., vriend, g., dijkstra, b.w., veltman, o.r., van den burg, b., venema, g., ulbrich-hofmann, r. & eijsink, v.g. (1997) j. biol. chem. 272, 11152-11156]. the study in guanidine hydrochloride of this enzyme and the respective wild-type enzyme allowed us to distingui ... | 2001 | 11422393 |
| e. coli rep oligomers are required to initiate dna unwinding in vitro. | e. coli rep protein is a 3' to 5' sf1 superfamily dna helicase which is monomeric in the absence of dna, but can dimerize upon binding either single-stranded or duplex dna. a variety of biochemical studies have led to proposals that rep dimerization is important for its helicase activity; however, recent structural studies of bacillus stearothermophilus pcra have led to suggestions that sf1 helicases, such as e. coli rep and e. coli uvrd, function as monomeric helicases. we have examined the que ... | 2001 | 11428893 |
| deduced amino-acid sequence of a calcium-free alpha-amylase from a strain of bacillus: implications from molecular modeling of high oxidation stability and chelator resistance of the enzyme. | alkaline alpha-amylase (amyk38) from the alkaliphilic bacillus sp. strain ksm-k38 is a unique enzyme in that it is highly chelator-resistant and oxidatively stable [hagihara, h., igarashi, k., hayashi, y., endo, k., ikawa-kitayama, k., ozaki, k., kawai, s. & ito, s. (2001) appl. environ. microbiol. 67, 1744-1750]. this enzyme was found to contain no ca and require na (or monovalent cations) for manifestation of activity. the nucleotide sequence of the gene for the novel enzyme was determined, an ... | 2001 | 11453991 |
| [evaluation of commercial uht milk and optimization of industrial process]. | from the nutrition al point of view milk is one of the most complete food in the diet of mammals. it contains nearly all the nutrients necessary to sustain life, but milk can deteriorate very easily, either by microbiological contamination or by biochemical reactions during processing and/or storage. the objective of this research study was to design a modified uht treatment to achieve commercial sterilization and maximize the stabilization of the heat-treated product during storage. to search f ... | 2000 | 11464666 |
| analysis of mutations at residues a2451 and g2447 of 23s rrna in the peptidyltransferase active site of the 50s ribosomal subunit. | on the basis of the recent atomic-resolution x-ray structure of the 50s ribosomal subunit, residues a2451 and g2447 of 23s rrna were proposed to participate directly in ribosome-catalyzed peptide bond formation. we have examined the peptidyltransferase and protein synthesis activities of ribosomes carrying mutations at these nucleotides. in escherichia coli, pure mutant ribosome populations carrying either the g2447a or g2447c mutations maintained cell viability. in vitro, the g2447a ribosomes s ... | 2001 | 11470897 |
| optimization of a thermostable lipase from bacillus stearothermophilus p1: overexpression, purification, and characterization. | an expression library was generated from a partial ncoi and hindiii digest of genomic dna from the thermophilic bacterium, bacillus stearothermophilus p1. the dna fragments were cloned into the expression vector pqe-60 and transformed into escherichia coli m15[ep4]. sequence analysis of a lipase gene showed an open reading frame of 1254 nucleotides coding a 29-amino-acid signal sequence and a mature sequence of 388 amino acids. the expressed lipase was isolated and purified to homogeneity in a s ... | 2001 | 11483000 |
| introduction of a (poly)histidine tag in l-lactate dehydrogenase produces a mixture of active and inactive molecules. | a (poly)histidine tag was fused to either the n- or the c-terminus of l-lactate dehydrogenase (ldh) of bacillus stearothermophilus to facilitate purification and immobilization of these enzymes. the c-terminally tagged enzyme displayed lower activity compared both to the wild-type and to the n-terminally tagged variant. the reason for this loss of activity was investigated by affinity chromatography of the enzymes on a 5'-amp-sepharose resin and by size-exclusion chromatography. the c-terminally ... | 2001 | 11488630 |
| influence of surface finish on the cleanability of stainless steel. | stainless steel for fabricating food processing equipment is available with various surface finishes. the objective of this research was to determine the effect of surface finish on cleanability. nine samples of stainless steel, type 304, from various manufacturers including no finish (hot rolled and pickled), #4 finish, 2b mechanical polished, and electropolished were tested. cleanability was assessed by using coupon samples soiled with either cultured milk inoculated with spores of bacillus st ... | 2001 | 11510656 |
| a cofactor-dependent phosphoglycerate mutase homolog from bacillus stearothermophilus is actually a broad specificity phosphatase. | the distribution of phosphoglycerate mutase (pgm) activity in bacteria is complex, with some organisms possessing both a cofactor-dependent and a cofactor-independent pgm and others having only one of these enzymes. although bacillus species contain only a cofactor-independent pgm, genes homologous to those encoding cofactor-dependent pgms have been detected in this group of bacteria, but in at least one case the encoded protein lacks significant pgm activity. here we apply sequence analysis, mo ... | 2001 | 11514674 |
| production of recombinant alpha-galactosidases in thermus thermophilus. | a thermus thermophilus selector strain for production of thermostable and thermoactive alpha-galactosidase was constructed. for this purpose, the native alpha-galactosidase gene (agat) of t. thermophilus th125 was inactivated to prevent background activity. in our first attempt, insertional mutagenesis of agat by using a cassette carrying a kanamycin resistance gene led to bacterial inability to utilize melibiose (alpha-galactoside) and galactose as sole carbohydrate sources due to a polar effec ... | 2001 | 11526023 |
| crystallization and preliminary x-ray analysis of a thermoalkalophilic lipase from bacillus stearothermophilus l1. | a thermoalkalophilic lipase from bacillus stearothermophilus l1 (l1 lipase) was crystallized in two different crystal forms using a low concentration of the enzyme and a calcium-exchange process. the first, needle-like, crystal form, which diffracts to about 3.5 a, belongs to the orthorhombic space group p2(1)2(1)2(1), with unit-cell parameters a = 67.84, b = 72.96, c = 104.41 a. the second, monoclinic, crystal form, which behaves better than the first form for crystallographic analyses, belongs ... | 2001 | 11526325 |
| changing the amino acid specificity of yeast tyrosyl-trna synthetase by genetic engineering. | in an attempt to generate mutant aminoacyl-trna synthetases capable of charging non-canonical amino acids, a series of yeast tyrosyl-trna synthetase (tyrrs) mutants was constructed by site-specific mutagenesis of putative active site residues, which were deduced by analogy with those of bacillus stearothermophilus tyrrs. among these mutants, one with the replacement of tyrosine at position 43 by glycine, "y43g," was found to be able to utilize several 3-substituted tyrosine analogues as substrat ... | 2001 | 11530018 |
| in vitro replication slippage by dna polymerases from thermophilic organisms. | replication slippage of dna polymerases is a potential source of spontaneous genetic rearrangements in prokaryotic and eukaryotic cells. here we show that different thermostable dna polymerases undergo replication slippage in vitro, during single-round replication of a single-stranded dna template carrying a hairpin structure. low-fidelity polymerases, such as thermus aquaticus (taq), high-fidelity polymerases, such as pyrococcus furiosus (pfu) and a highly thermostable polymerase from pyrococcu ... | 2001 | 11554789 |
| efficacy of chemical sterilization and storage conditions of gutta-percha cones. | the objective of the present study was to assess the efficacy of 2.5% sodium hypochlorite and 2.2% glutaraldehyde ('cidex') as sterilizing agents for gutta-percha cones. the efficacy of storage of gutta-percha cones in the presence or absence of paraformaldehyde was also evaluated. | 2001 | 11556509 |
| the effect of changing the hydrophobic s1' subsite of thermolysin-like proteases on substrate specificity. | the hydrophobic s1' subsite is one of the major determinants of the substrate specificity of thermolysin and related m4 family proteases. in the thermolysin-like protease (tlp) produced by bacillus stearothermophilus (tlp-ste), the hydrophobic s1' subsite is mainly formed by phe130, phe133, val139 and leu202. in the present study, we have examined the effects of replacing leu202 by smaller (gly, ala, val) and larger (phe, tyr) hydrophobic residues. the mutational effects showed that the wild-typ ... | 2001 | 11559368 |
| isolation of an individual allosteric interaction in tetrameric phosphofructokinase from bacillus stearothermophilus. | phosphofructokinase from bacillus stearothermophilus (bspfk) is a model allosteric enzyme system in which the interactions between substrates and allosteric effectors have been extensively studied. however, the oligomeric nature of bspfk has made it difficult to determine the molecular basis of the allosteric regulation because of the multitude of different types of heterotropic and homotropic interactions that are possible between the four active sites and four allosteric sites in the native te ... | 2001 | 11560513 |
| on the relationship between protein stability and folding kinetics: a comparative study of the n-terminal domains of rnase hi, e. coli and bacillus stearothermophilus l9. | there is currently a great deal of interest in proteins that fold in a single highly cooperative step. particular attention has been focused on elucidating the factors that govern folding rates of simple proteins. recently, the topology of the native state has been proposed to be the most important determinant of their folding rates. here we report a comparative study of the folding of three topologically equivalent proteins that adapt a particularly simple alpha/beta fold. the folding kinetics ... | 2001 | 11563917 |
| modeling and analysis of the structure of the thermostable catechol 2,3-dioxygenase from bacillus stearothermophilus. | the three-dimensional structure of thermostable catechol 2,3-dioxygenase(tc230) from bacillus stearothermophilus has been modeled basing on the known x-ray structure of catechol 2,3-dioxygenase(metapyrocatechase) from pseudomonas putida mt-2, using computer graphics energy minimization techniques. the rationality of the resulting model was validated by ramachandran plot and profile-3d. the structure-functionally important residues, such as m++ binding residues and the substrate binding residues, ... | 2001 | 11565853 |
| glycerol dehydrogenase. structure, specificity, and mechanism of a family iii polyol dehydrogenase. | bacillus stearothermophilus glycerol dehydrogenase (glydh) (glycerol:nad(+) 2-oxidoreductase, ec 1.1.1.6) catalyzes the oxidation of glycerol to dihydroxyacetone (1,3-dihydroxypropanone) with concomitant reduction of nad(+) to nadh. analysis of the sequence of this enzyme indicates that it is a member of the so-called iron-containing alcohol dehydrogenase family. despite this sequence similarity, glydh shows a strict dependence on zinc for activity. on the basis of this, we propose to rename thi ... | 2001 | 11566129 |
| toxic effects of tamoxifen on the growth and respiratory activity of bacillus stearothermophilus. | the anticancer drug tamoxifen (tam) is used as first line therapy in breast cancer. although tamoxifen is usually considered an estrogen antagonist, several studies suggest alternative mechanisms of action. bacillus stearothermophilus has been used as a model to clarify the antiproliferative action of tamoxifen putatively related with drug-membrane interaction. according to previous data, tam induces perturbation of membrane structure along with impairment of bacterial growth. the aim of this wo ... | 2001 | 11566553 |
| crystal structure of staphylococcus aureus tyrosyl-trna synthetase in complex with a class of potent and specific inhibitors. | sb-219383 and its analogues are a class of potent and specific inhibitors of bacterial tyrosyl-trna synthetases. crystal structures of these inhibitors have been solved in complex with the tyrosyl-trna synthetase from staphylococcus aureus, the bacterium that is largely responsible for hospital-acquired infections. the full-length enzyme yielded crystals that diffracted to 2.8 a resolution, but a truncated version of the enzyme allowed the resolution to be extended to 2.2 a. these inhibitors not ... | 2001 | 11567092 |
| evidence for a novel role of copper-zinc superoxide dismutase in zinc metabolism. | the lys7 gene in saccharomyces cerevisiae encodes a protein (yccs) that delivers copper to the active site of copper-zinc superoxide dismutase (cuzn-sod, a product of the sod1 gene). in yeast lacking lys7 (lys7delta), the sod1 polypeptide is present but inactive. mutants lacking the sod1 polypeptide (sod1delta) and lys7delta yeast show very similar phenotypes, namely poor growth in air and aerobic auxotrophies for lysine and methionine. here, we demonstrate certain phenotypic differences between ... | 2001 | 11581253 |
| a microwave-powered sterilizable interface for aseptic access to bioreactors that are vulnerable to microbial contamination. | novel methods and apparatus that employ the rapid heating characteristics of microwave irradiation to facilitate the aseptic transfer of nutrients, products, and other materials between microbially sensitive systems and the external environment are described. the microwave-sterilizable access port (msap) consists of a 600-w magnetron emitting at a frequency of 2.45 ghz, a sterilization chamber with inlet and outlet flow lines, and a specimen transfer interface. energy is routed to the sterilizat ... | 2001 | 11587573 |
| the efficacy of chemical agents in cleaning and disinfection programs. | due to the growing number of outbreaks of infection in hospital nurseries, it becomes essential to set up a sanitation program that indicates that the appropriate chemical agent was chosen for application in the most effective way. | 2001 | 11591223 |
| a rapid assay for detecting sulfonamides in tissues of slaughtered animals. | governments regulate antimicrobial residues in slaughtered animals with surveillance programs for detecting drugs in food-producing animals. although initial screening bioassay systems are recognized for their sensitivities to antimicrobial drug groups, none are sensitive to sulfonamides at or near the maximum residue levels (mrls) in the codex alimentarious. we have developed a sulfonamide-sensitive rapid assay using bacillus stearothermophilus inoculated pm indicator agar containing bromcresol ... | 2001 | 11601706 |
| modeling the combined effect of ph and temperature on the heat resistance of bacillus stearothermophilus spores heated in a multicomponent food extract. | the combined effect of ph and temperature on the heat resistance of bacillus stearothermophilus spores heated in an extract of complex food was studied. the results showed that, in general, reducing the ph reduced the heat resistance of the spores. similarly, the value for the d parameter in the nonacidified extract was between 30 and 70% lower than the one obtained with double-distilled water. this result once again shows the importance of the substrate in inactivation studies of microorganisms ... | 2001 | 11601720 |
| modulation of the regioselectivity of a bacillus alpha-galactosidase by directed evolution. | alpha-galactosidase agab of bacillus stearothermophilus was subjected to directed evolution in an effort to modify its regioselectivity. the wild-type enzyme displays a major 1,6 and minor 1,3 regioselectivity. we used random mutagenesis and staggered extension process (step) to obtain mutant enzymes displaying modified regioselectivity. we developed a screening procedure allowing first the elimination of agab mutants bearing the 1,6 regioselectivity and secondly the selection of those retaining ... | 2001 | 11602805 |
| density functional and electrostatic calculations of manganese superoxide dismutase active site complexes in protein environments. | density functional and electrostatic methods have been applied to calculate active site geometries and the redox potential of manganese superoxide dismutase (mnsod). the initial active site clusters were built up by including only first-shell side chain ligands and then augmented by second-shell ligands. the density functional optimized mn-ligand bond lengths for the reduced complexes in general compared fairly well with protein crystallography data; however, large deviations for calculated mn-o ... | 1999 | 11670865 |
| the product of the cysk gene of bacillus stearothermophilus v mediates potassium tellurite resistance in escherichia coli. | the nucleotide sequence of a 4,539 bp fragment of bacillus stearothermophilus v mediating tellurite resistance in escherichia coli was determined. four orfs of more than 150 amino acids encoding polypeptides of 244, 258, 308, and 421 residues were found in the restriction fragment. e. coli cells harboring a recombinant plasmid containing the ter determinant express, when challenged with tellurite, a 32 kda protein with an amino terminal sequence identical to the ten first residues of the 308 orf ... | 2001 | 11685509 |
| biodecontamination of animal rooms and heat-sensitive equipment with vaporized hydrogen peroxide. | common methods used to decontaminate and disinfect laboratory animal areas are difficult to standardize, labor-intensive, and potentially hazardous for staff members and the environment. as an alternative to traditional methods, we tested fumigation with vaporized hydrogen peroxide by using the vhp 1000 biodecontamination system. the design of our air-conditioning system allowed the connection of the generator to any animal room by using the ventilation piping, thus forming a closed circuit. a 3 ... | 2001 | 11703051 |
| ultraviolet disinfection with a novel microwave-powered device. | to evaluate the antimicrobial efficacy of a novel u.v. beaker, powered in a domestic microwave oven. | 2001 | 11722655 |
| controlling residual dipolar couplings in high-resolution nmr of proteins by strain induced alignment in a gel. | water-soluble biological macromolecules can be weakly aligned by dissolution in a strained, hydrated gel such as cross-linked polyacrylamide, an effect termed 'strain-induced alignment in a gel' (sag). sag induces nonzero nuclear magnetic dipole-dipole couplings that can be measured in high-resolution nmr spectra and used as structural constraints. the dependence of experimental 15n-1h dipolar couplings extracted from two-dimensional heteronuclear single quantum coherence (hsqc) spectra on sever ... | 2001 | 11727977 |
| studies on bacillus stearothermophilus. part 1. transformation of progesterone to a new metabolite 9,10-seco-4-pregnene-3,9,20-trione. | when bacillus stearothermophilus, a thermophilic bacterium isolated from the kuwaiti desert, was incubated with exogenous progesterone for 24 h, three monohydroxylated metabolites were produced. 20alpha-hydroxyprogesterone was the major metabolite produced in 60.8 relative percentage yield. the other two monohydroxylated metabolites were identified as 6beta-hydroxyprogesterone and the rare 6alpha-hydroxyprogesterone in 21.0 and 13.6 relative percentage yields, respectively. a new metabolite 9,10 ... | 2001 | 11738559 |
| the ptsi gene encoding enzyme i of the phosphotransferase system of corynebacterium glutamicum. | the phosphoenolpyruvate:carbohydrate phosphotransferase system (pts) is widespread among bacteria where it mediates carbohydrate uptake and often serves in carbon control. here we present cloning and analysis of the monocistronic ptsi gene of corynebacterium glutamicum r, which encodes pts enzyme i (ei). ei catalyzes the first reaction of pts and the reported ptsi was shown to complement the corresponding defect in escherichia coli. the deduced 59.2-kda ei of 564 amino acids shares more than 50% ... | 2001 | 11741338 |
| the surface layer (s-layer) glycoprotein of geobacillus stearothermophilus nrs 2004/3a. analysis of its glycosylation. | geobacillus stearothermophilus nrs 2004/3a possesses an oblique surface layer (s-layer) composed of glycoprotein subunits as the outermost component of its cell wall. in addition to the elucidation of the complete s-layer glycan primary structure and the determination of the glycosylation sites, the structural gene sgse encoding the s-layer protein was isolated by polymerase chain reaction-based techniques. the open reading frame codes for a protein of 903 amino acids, including a leader sequenc ... | 2002 | 11741945 |
| expression, purification, crystallization and preliminary crystallographic analysis of a thermostable lipase from bacillus stearothermophilus p1. | the gene encoding a thermostable lipase secreted by bacillus stearothermophilus p1 has been cloned and overexpressed in escherichia coli. the recombinant lipase was purified to homogeneity using ammonium sulfate precipitation, anion-exchange chromatography (poros 20 hq) and sephacryl s-200hr. the molecular mass was shown to be 43 209 da by mass spectrometry. crystals suitable for x-ray diffraction analysis were obtained by the hanging-drop method of vapour diffusion with ammonium sulfate as the ... | 2002 | 11752807 |
| reclassification of saccharococcus caldoxylosilyticus as geobacillus caldoxylosilyticus (ahmad et al. 2000) comb. nov. | a polyphasic study was performed on five thermophilic strains belonging to the genus bacillus, isolated from soil of different geographical areas. 16s rrna gene sequence analysis placed these isolates in rna group 5, with saccharococcus caldoxylosilyticus and [bacillus] thermoglucosidasius being the closest phylogenetic neighbours. the type species of saccharococcus, saccharococcus thermophilus, was only moderately related to these two species and the novel isolates. dna-dna hybridization studie ... | 2001 | 11760948 |
| [use of and verification with biological indicators in sterilizers belonging to dentistry surgeons from san luis potosi, mexico]. | to assess the utilization of sterilizing equipment used by dentists, and verification of sterilization using biological indicators. | 2001 | 11763693 |
| disinfection/sterilization of extracted teeth for dental student use. | extracted human teeth are used in many preclinical courses. while there has been no report of disease transmission with extracted teeth, sterilization of teeth used in the teaching laboratory should be a concern. the purpose of this study was to determine the effectiveness of different sterilization/disinfection methods of extracted human teeth using bacillus stearothermophilus, a bacteria resistant to heat and frequently used to test sterilizers. in this study, 110 extracted molars with no cari ... | 2001 | 11765875 |
| joint refinement as a tool for thorough comparison between nmr and x-ray data and structures of hu protein. | joint refinement, i.e., the simultaneous refinement of a structure against both nuclear magnetic resonance (nmr) spectroscopic and x-ray crystallographic data, was performed on the hu protein from bacillus stearothermophilus (hubst). the procedure was aimed at investigating the compatibility of the two data sets and at identifying conflicting information. wherever important differences were found, such as peptide flips in the main-chain conformation, the data were further analyzed to find the ca ... | 2001 | 11775740 |
| a catalase-peroxidase from a newly isolated thermoalkaliphilic bacillus sp. with potential for the treatment of textile bleaching effluents. | a new thermoalkaliphilic bacterium was isolated from a textile wastewater drain and identified as a new bacillus sp. (bacillus sf). because of its high ph stability and thermostability, a catalase-peroxidase (cp) from this strain has potential for the treatment of textile bleaching effluents. the cp from bacillus sf was purified to more than 70.3-fold homogeneity using fractionated ammonium sulfate precipitation, hydrophobic interaction, and anion-exchange and gel-filtration chromatography. the ... | 2001 | 11778844 |
| compact form of dna induced by dna-binding protein hu. | interaction of dna-binding protein hu from bacillus stearothermophilus (hubst) with coliphage t2 dna was investigated by means of a single-duplex dna chain visualization method using fluorescence microscopy. fluorescence microscopic images of coliphage t2 dna molecules were observed as a function of hubst concentration. the average fluorescence image size of t2 dna decreased with increase in hubst concentration to a size comparable to that of a dna globule induced by polyethylene glycol (peg) an ... | 2002 | 11779206 |
| transcription regulation in thermophilic bacteria: high resolution contact probing of bacillus stearothermophilus and thermotoga neapolitana arginine repressor-operator interactions. | arginine-mediated regulation is remarkably well conserved in very divergent bacteria, and shows a number of unusual features that distinguish arginine regulation from other transcriptional control mechanisms. the arginine repressor subunit consists of a basic n-terminal dna-binding domain, which belongs to the winged helix-turn-helix family, connected through a flexible linker to an acidic c-terminal domain responsible for binding of arginine and assembly of the high-affinity holohexamer, which ... | 2002 | 11786010 |
| the anti-trp rna-binding attenuation protein (anti-trap), at, recognizes the tryptophan-activated rna binding domain of the trap regulatory protein. | in bacillus subtilis, the trp rna-binding attenuation protein (trap) regulates expression of genes involved in tryptophan metabolism in response to the accumulation of l-tryptophan. tryptophan-activated trap negatively regulates expression by binding to specific mrna sequences and either promoting transcription termination or blocking translation initiation. conversely, the accumulation of uncharged trna(trp) induces synthesis of an anti-trap protein (at), which forms a complex with trap and inh ... | 2002 | 11786553 |
| sporicidal action of ozone and hydrogen peroxide: a comparative study. | elimination of contaminating spores on packaging materials and food-contact surfaces remains a challenge to the food industry. hydrogen peroxide and chlorine are the most commonly used sanitizers to eliminate these contaminants, and ozone was recommended recently as an alternative. hence, we compared the sporicidal action of ozone and hydrogen peroxide against selected foodborne spores of bacillus spp. under identical treatment conditions, 11 microg/ml aqueous ozone decreased spore counts by 1.3 ... | 2001 | 11789930 |
| characterization of an s-layer glycoprotein produced in the course of s-layer variation of bacillus stearothermophilus atcc 12980 and sequencing and cloning of the sbsd gene encoding the protein moiety. | the cell surface of bacillus stearothermophilus atcc 12980 is completely covered by an oblique lattice which consists of the s-layer protein sbsc. on sds-polyacrylamide gels, the mature s-layer protein migrates as a single band with an apparent molecular mass of 122 kda. during cultivation of b. stearothermophilus atcc 12980 at 67 degrees c instead of 55 degrees c, a variant developed that had a secondary cell wall polymer identical to that of the wild-type strain, but it carried an s-layer glyc ... | 2001 | 11797047 |
| morpho-structural variations of bacterial spores after treatment in steam vacuum assisted autoclave. | this study intended to verify, through microbiological techniques and tem investigations, the killing of bacterial spores after treatment in steam autoclave, and to propose strictly morphological considerations about the target of this sterilisation process. autoclave is the most common device for sterilising instruments in order to prevent cross infections in dental offices. the autoclave efficiency has been improved in the last years and part of this improvement is related to both a better and ... | 1999 | 11799742 |
| samy, a novel mammalian reporter gene derived from bacillus stearothermophilus alpha-amylase. | the bacillus stearothermophilus alpha-amylase (amys) is a heat-stable monomeric exoenzyme which catalyses random hydrolysis of 1,4-alpha-glucosidic linkages in polyglucosans. the bacillus alpha-amylase was engineered for use as an intracellular (amys(delta s)) as well as a secreted reporter protein (samy; secreted alpha-amylase) in mammalian cells. the 5' end of amys containing the prokaryotic secretion signal was either deleted (amys(delta s)) or replaced by a murine immunoglobulin secretion si ... | 2002 | 11814674 |
| structure and mechanism of action of a cofactor-dependent phosphoglycerate mutase homolog from bacillus stearothermophilus with broad specificity phosphatase activity. | the crystal structure of bacillus stearothermophilus phoe (originally termed yhfr), a broad specificity monomeric phosphatase with a molecular mass of approximately 24 kda, has been solved at 2.3 a resolution in order to investigate its structure and function. phoe, already identified as a homolog of a cofactor-dependent phosphoglycerate mutase, shares with the latter an alpha/beta/alpha sandwich structure spanning, as a structural excursion, a smaller subdomain composed of two alpha-helices and ... | 2002 | 11827481 |
| catalytic activities of intracellular dimeric neopullulanase on cyclodextrin, acarbose and maltose. | multi-substrate specificity of neopullulanase towards cyclodextrin, acarbose and maltose was investigated using a clone originating from bacillus stearothermophilus ima6503. the enzyme purified from escherichia coli harbouring the corresponding npla gene hydrolysed beta-cyclodextrin (beta-cd) to maltose and glucose. it exhibited substrate preference for beta-cd, starch and pullulan in the proportions of 10.4:1.2:1. the enzyme not only hydrolysed acarbose, an alpha-amylase inhibitor, to a pseudot ... | 2002 | 11834127 |
| crystal structure of the bse634i restriction endonuclease: comparison of two enzymes recognizing the same dna sequence. | crystal structures of type ii restriction endonucleases demonstrate a conserved common core and active site residues but diverse structural elements involved in dna sequence discrimination. comparative structural analysis of restriction enzymes recognizing the same nucleotide sequence might therefore contribute to our understanding of the structural diversity of specificity determinants within restriction enzymes. we have solved the crystal structure of the bacillus stearothermophilus restrictio ... | 2002 | 11842098 |
| a functional interaction between the putative primosomal protein dnai and the main replicative dna helicase dnab in bacillus. | in gram negative escherichia coli there are two well-characterised primosomal assembly processes, the pria- and dnaa-mediated cascades. the presence of pria and dnaa proteins in gram positive bacillus spp. supports the assumption that both the pria- and dnaa-mediated primosomal assembly cascades also operate in these organisms. however, the lack of sequence homology between the rest of the primosomal proteins indicates significant differences between these two bacterial species. central to the p ... | 2002 | 11842108 |
| interaction of the bacillus stearothermophilus ribosomal protein s15 with its 5'-translational operator mrna. | the bacillus stearothermophilus ribosomal protein s15 (bs15) binds both a three-helix junction in the central domain of 16 s ribosomal rna and its cognate mrna. native gel mobility-shift assays show that bs15 interacts specifically and with high affinity to the 5'-untranslated region (5'-utr) of this cognate mrna with an apparent dissociation constant of 3(+/-0.3) nm. in order to localize the structural elements that are essential for bs15 recognition, a series of deletion mutants of the full co ... | 2001 | 11846555 |
| cloning and expression analysis of the 28 kda protein from lactobacillus delbrueckii subsp. lactis atcc 4797 hypothesized to influence lactacin b production. | a cell wall-associated lactacin b inducer protein (ip) was purified from lactobacillus delbrueckii subsp. lactis atcc 4797 (lact. lactis) by chromatofocusing and gel filtration hplc (barefoot et al. 1994). | 2001 | 11851815 |
| identification and properties of type i-signal peptidases of bacillus amyloliquefaciens. | the use of bacillus amyloliquefaciens for enzyme production and its exceptional high protein export capacity initiated this study where the presence and function of multiple type i signal peptidase isoforms was investigated. in addition to type i signal peptidases sips(ba) [meijer, w.j.j., de jong, a., bea, g., wisman, a., tjalsma, h., venema, g., bron, s. & van dijl, j.m. (1995) mol. microbiol. 17, 621-631] and sipt(ba) [hoang, v. & hofemeister, j. (1995) biochim. biophys. acta 1269, 64-68] whi ... | 2002 | 11856304 |