| molecular and enzymological evidence for two classes of fumarase in bacillus stearothermophilus (var. non-diastaticus). | the gene (fumabst) encoding an oxygen-labile fumarase of bacillus stearothermophilus has been cloned and sequenced. the structural gene (1542 bp) encodes a product (fumabst) of m(r) 56,788 containing 514 amino acid residues. the amino acid sequence is 23% identical (37% similar) to fuma and fumb, the labile [4fe-4s]-containing fumarases (class i enzymes) of escherichia coli. it exhibits no significant similarity to fumc and citg, the stable fumarases (class ii enzymes) of e. coli and bacillus su ... | 1993 | 8473853 |
| identification of a putative infc-rpmi-rplt operon flanked by long inverted repeats in mycoplasma fermentans (incognitus strain). | a specific 1542-bp dna fragment was amplified from mycoplasma fermentans (incognitus strain) using a unique 23-nucleotide (nt) synthetic deoxyribonucleotide (oligo) (5'-tccaaaaagtccggaatttgggg) as the primer pair in the polymerase chain reaction (pcr). the 23-nt sequence is part of the 29-bp terminal inverted repeat (ir) which forms the left potential stem-and-loop (s&l) structure of the previously identified m. fermentans insertion-sequence(is)-like genetic element [hu et al., gene 93 (1990) 67 ... | 1993 | 8486291 |
| thermostable farnesyl diphosphate synthase of bacillus stearothermophilus: molecular cloning, sequence determination, overproduction, and purification. | the structural gene for thermostable farnesyl diphosphate synthase from bacillus stearothermophilus was cloned, sequenced, and overexpressed in escherichia coli cells. a 1,260-nucleotide sequence of the cloned fragment was determined. this sequence specifies an open reading frame of 891 nucleotides for farnesyl diphosphate synthase. the deduced amino acid sequence shows a 42% similarity with that of e. coli fpp synthase [fujisaki et al. (1990) j. biochem. 108, 995-1000]. comparison with prenyltr ... | 1993 | 8486607 |
| field trial of four cowside antibiotic-residue screening tests. | four commercially available screening tests for antibiotic residues in milk were evaluated for their ability to correctly identify the antibiotic status of cows. a field trial, which included 199 cows from 2 herds, was conducted. sensitivity, specificity, and likelihood ratios for a positive test result were calculated by using the bacillus stearothermophilus var calidolactis disk assay as the reference test. the relationship of risk factors to the probability of a false-positive result for each ... | 1993 | 8496080 |
| reaction of modified and unmodified trna(tyr) substrates with tyrosyl-trna synthetase (bacillus stearothermophilus). | three species of trna(tyr) have been examined as substrates for the transfer reaction of the tyrosyl-trna synthetase (tyrrs) from bacillus stearothermophilus: escherichia coli trna(tyr), b. stearothermophilus trna(tyr) expressed in e. coli, and b. stearothermophilus trna(tyr) that has been transcribed in vitro. the binding of the first two substrates to tyrrs may be readily monitored by stopped-flow studies of tryptophan fluorescence to give the rate and equilibrium constants. the in vitro-trans ... | 1993 | 8499435 |
| use of binding energy in catalysis: optimization of rate in a multistep reaction. | the role of binding energy in optimizing the overall rate of catalysis by the tyrosyl-trna synthetase from bacillus stearothermophilus has been investigated by measuring the rate constants for transfer of tyrosine from engineered mutants to trna. the residues chosen for mutation are those that were previously identified as binding tyrosyl adenylate and contributing to the rate constant for its formation. it was previously found that tighter binding of the tyrosyl adenylate was accompanied by an ... | 1993 | 8499436 |
| bsp1407i, a restriction endonuclease from bacillus stearothermophilus, which recognizes novel palindromic sequence 5'-t decreases gtaca-3'. | | 1993 | 8506146 |
| molecular cloning, sequence analysis and expression of the gene for catalase-peroxidase (cpea) from the photosynthetic bacterium rhodobacter capsulatus b10. | the gene encoding catalase-peroxidase was cloned from chromosomal dna of rhodobacter capsulatus b10. the nucleotide sequence of a 3.7-kb saci-hindiii fragment, containing the catalase-peroxidase gene (cpea) and its flanking regions were determined. a 1728-bp open reading frame, coding for 576 amino acid residues (molecular mass 61516 da) of the enzyme, was observed. a shine-dalgarno sequence was found 5 bp upstream from the translational start site. the deduced amino acid sequence coincides with ... | 1993 | 8508796 |
| saccharomyces cerevisiae cytoplasmic tyrosyl-trna synthetase gene. isolation by complementation of a mutant escherichia coli suppressor trna defective in aminoacylation and sequence analysis. | exploiting differences in trna recognition between prokaryotic and eukaryotic tyrosyl-trna synthetases (tyrrss), we have isolated the gene for the cytoplasmic tyrrs of saccharomyces cerevisiae by functional complementation in escherichia coli of a mutant e. coli trna. the trna, derived from the e. coli initiator trna with changes to allow suppression of amber termination codons, is poorly aminoacylated in e. coli and hence, is only a weak amber suppressor. the same trna functions as a good suppr ... | 1993 | 8509419 |
| effect of sterilization on contaminated sponges. | sponges are routinely used as a storage medium for endodontic files during clinical practice; however, very little research has been done to determine the effectiveness of sterilization procedures for these contaminated sponges. the purpose of this in vitro study was to determine the efficacy of chemical vapor sterilizers (chemiclaves), steam pressure sterilizers (autoclaves), and dry heat sterilizers on laboratory contaminated sponges. four different types of sponges were used in this study: a ... | 1993 | 8509738 |
| identifying the mechanism of protein loop closure: a molecular dynamics simulation of the bacillus stearothermophilus ldh loop in solution. | the 'loop' involving residues 98-110 in bacillus stearothermophilus lactate dehydrogenase (bsldh) is of great interest as substrate-induced 'loop' closure is thought to be rate-limiting and essential in catalyzing the reaction and in determining substrate specificity. consequently, we have explored the mechanism underlying 'loop' opening in bsldh through a molecular dynamics simulation at high temperature (1000 k) in the presence of explicit solvent, starting from the x-ray structure of bsldh co ... | 1995 | 8532681 |
| a general method for the consecutive integration of single copies of a heterologous gene at multiple locations in the bacillus subtilis chromosome by replacement recombination. | we have devised a two-step procedure by which multiple copies of a heterologous gene can be consecutively integrated into the bacillus subtilis 168 chromosome without the simultaneous integration of markers (antibiotic resistance). the procedure employs the high level of transformability of b. subtilis 168 strains and makes use of the observation that thymine-auxotrophic mutants of b. subtilis are resistant to the folic acid antagonist trimethoprim (tmpr), whereas thymine prototrophs are sensiti ... | 1995 | 8534091 |
| reddish escherichia coli cells caused by overproduction of bacillus stearothermophilus uroporphyrinogen iii methylase: cloning, sequencing, and expression of the gene. | during shotgun cloning of an amylase gene, we found a transformant of escherichia coli with a reddish color. the transformant produced highly water-soluble red pigments the molecular masses of which were less than 3000. the plasmid harbored by the transformant contained a dna fragment derived from a strain of bacillus stearothermophilus. truncation of the insert dna showed that an 1.1-kbp sau 3a-sali fragment was responsible for the reddish colony. an open reading frame was found in the nucleoti ... | 1995 | 8534969 |
| cloning and sequencing of a gene encoding d-aminoacylase from alcaligenes xylosoxydans subsp. xylosoxydans a-6 and expression of the gene in escherichia coli. | the gene encoding the d-aminoacylase of alcaligenes xylosoxydans subsp. xylosoxydans a-6 (alcaligenes a-6) was cloned and its complete nucleotide sequence was identified. the d-aminoacylase structural gene consists of 1452 nucleotides and encodes 484 amino acid residues. the molecular weight of d-aminoacylase was calculated to be 51,918. this value agreed well with the apparent molecular weight of 52,000 found for the purified enzyme from alcaligenes a-6 by sodium dodecyl sulfate (sds)-polyacryl ... | 1995 | 8541651 |
| [cloning and expression of the bacillus stearothermophilus neutral proteinase gene in bacillus subtilis cells]. | gene of thermostable bacillus stearothermophilus metalloprotease was cloned and expressed in mesophilic bacillus subtilis cells. it was demonstrated that this gene is closely related by its restriction map to b. stearothermophilus t metalloprotease gene cloned earlier. thermostability level and thermal optimum of activity of metalloprotease, the product of cloned gene expression, were estimated. | 1995 | 8552054 |
| escherichia coli tryptophanyl-trna synthetase mutants selected for tryptophan auxotrophy implicate the dimer interface in optimizing amino acid binding. | tryptophan auxotrophs of escherichia coli in which mutations were mapped to the trps locus (encoding tryptophanyl-trna synthetase) have been previously isolated. we have investigated the tryptophanyl-trna synthetase (trprs) purified from six auxotrophic strains for changes in amino acid activation and aminoacylation. steady-state kinetic analyses show that these mutant trprs proteins have increases in the apparent km for tryptophan, decreases in turnover number, or both, without significant chan ... | 1996 | 8555191 |
| a glyceraldehyde-3-phosphate dehydrogenase homolog in borrelia burgdorferi and borrelia hermsii. | a polyreactive monoclonal antibody recognized a 38.5-kda polypeptide with amino-terminal sequence identity to conserved regions of glyceraldehyde-3-phosphate dehydrogenase (gapdh) in borrelia burgdorferi, the lyme disease agent, and borrelia hermsii, an agent of american relapsing fever. this monoclonal antibody also recognized gapdh from other pathogenic spirochetes and other prokaryotes and eukaryotes as well. gapdh activity was detected in sonicates of both b. burgdorferi and b. hermsii but n ... | 1996 | 8557349 |
| disordered c-terminal domain of tyrosyl transfer-rna synthetase: evidence for a folded state. | the c-terminal domain (residues 320 to 419) of tyrosyl-trna synthetase from bacillus stearothermophilus (bst-tyrrs) is necessary for the binding of trna(tyr) but disordered in the crystal structure. four different criteria showed that the isolated c-terminal domain of bst-tyrrs was at least partially folded in solution. its spectrum of circular dichroism was compatible with a high content of secondary structure elements (56% of its residues) and these structural elements disappeared in 7.5 m ure ... | 1996 | 8568859 |
| glycine-15 in the bend between two alpha-helices can explain the thermostability of dna binding protein hu from bacillus stearothermophilus. | on the basis of sequence comparison of thermophilic and mesophilic dna binding protein hus, bacillus stearothermophilus dna binding protein hu (bsthu) seems to gain thermostability with a change in amino acid residues present on the molecular surface. to evaluate the contribution of exchange of each amino acid to the thermostability of bsthu, we constructed three mutants, bsthu-t13a (thr13 to ala), bsthu-g15e (gly15 to glu), and bsthu-t33l (thr33 to leu), in which the amino acids in bsthu were c ... | 1996 | 8573574 |
| sequence analysis of flanking regions of the pfoa gene of clostridium perfringens: beta-galactosidase gene (pbg) is located in the 3'-flanking region. | the 3'-flanking region of the perfringolysin o (theta-toxin) gene (pfoa) of clostridium perfringens was analyzed by chromosome walking. a total of 5,363 bp of the downstream region of the pfoa gene was sequenced and four open reading frames were found. orf54 and orf80 were found to be homologous to genes coding for membrane-bound transporter proteins of other bacteria and the beta-galactosidase gene (bgab) of bacillus stearothermophilus, respectively. orf80 was named the pbg gene. clones which s ... | 1995 | 8577281 |
| crystal structure of recombinant triosephosphate isomerase from bacillus stearothermophilus. an analysis of potential thermostability factors in six isomerases with known three-dimensional structures points to the importance of hydrophobic interactions. | the structure of the thermostable triosephosphate isomerase (tim) from bacillus stearothermophilus complexed with the competitive inhibitor 2-phosphoglycolate was determined by x-ray crystallography to a resolution of 2.8 a. the structure was solved by molecular replacement using xplor. twofold averaging and solvent flattening was applied to improve the quality of the map. active sites in both the subunits are occupied by the inhibitor and the flexible loop adopts the "closed" conformation in ei ... | 1995 | 8580851 |
| genes and enzymes of the acetyl cycle of arginine biosynthesis in corynebacterium glutamicum: enzyme evolution in the early steps of the arginine pathway. | a cluster of arginine biosynthetic genes of corynebacterium glutamicum atcc 13032, comprising argj, argb and argd as well as part of argc and argf, has been cloned by heterologous complementation of an escherichia coli arge mutant. the gene order has been established as argcjbdf by sequencing the entire 4.4 kb cloned dna fragment. the c. glutamicum argb gene can be transcribed in e. coli cells from an internal promoter located in the coding part of the preceding argj gene, whereas transcription ... | 1996 | 8581175 |
| phylogenetic analysis of the putative phosphorylation domain in the pyruvate kinase of bacillus stearothermophilus. | all sequenced phosphoenolpyruvate synthases (pps), pyruvate:phosphate dikinases (ppdk) and enzymes i (ei) of the phosphoenolpyruvate:sugar phosphotransferase system comprise the pep family. linked to the c terminus of the sequenced pyruvate kinase from bacillus stearothermophilus (pkbst) is a domain that is homologous to the putative phosphorylation domains of pep family enzymes. we report sequence and phylogenetic analyses that lead to the following conclusions: (1) the phosphorylation domain o ... | 1995 | 8584793 |
| overproduction, purification and characterization of groes and groel from thermophilic bacillus stearothermophilus. | to facilitate purification of the two chaperonins groes and groel encoded by the thermophilic bacillus stearothermophilus, an escherichia coli strain was constructed in which the geoesl operon was replaced by that of b. stearothermophilus. this strain is perfectly viable, demonstrating that the b. stearothermophilus operon is functionally interchangeable with that of e. coli. to increase the amount of groes, the groes gene was fused to an iptg-inducible promoter. both proteins groes and groel, w ... | 1995 | 8586266 |
| the suitability of a monofunctional reagent of an undecagold cluster for phasing data collected from the large ribosomal subunits from bacillus stearothermophilus. | an electron density map of the large ribosomal subunit from bacillus stearothermophilus was obtained at 26 a resolution by single isomorphous replacement (sir) from a derivative formed by specific quantitative labeling with a dense undecagold cluster. for derivatization, a monofunctional reagent of this cluster was bound to a sulfhydryl group of a purified ribosomal protein, which was in turn reconstituted with core particles of a mutant lacking this protein. the native, mutated, and derivatized ... | 1995 | 8589246 |
| cation-selectivity of the l-glutamate transporters of escherichia coli, bacillus stearothermophilus and bacillus caldotenax: dependence on the environment in which the proteins are expressed. | l-glutamate transport by the h(+)-glutamate and na(+)-glutamate symport proteins of escherichia coli k-12 (gltpec and gltsec, respectively) and the na(+)-h(+)-glutamate symport proteins of bacillus stearothermophilus (glttbs) and bacillus caldotenax (glttbc) was studied in membrane vesicles derived from cells in which the proteins were either homologously or heterologously expressed. substrate and inhibitor specificity studies indicate that gltpec, glttbs and glttbc fall into the same group of t ... | 1995 | 8596452 |
| dynamics in oxygen-induced changes in s-layer protein synthesis from bacillus stearothermophilus pv72 and the s-layer-deficient variant t5 in continuous culture and studies of the cell wall composition. | stable synthesis of the hexagonally ordered (p6) s-layer protein from the wild-type strain of bacillus stearothermophilus pv72 could be achieved in continuous culture on complex medium only under oxygen-limited conditions when glucose was used as the sole carbon source. depending on the adaptation of the wild-type strain to low oxygen supply, the dynamics in oxygen-induced changes in s-layer protein synthesis was different when the rate of aeration was increased to a level that allowed dissimila ... | 1996 | 8606191 |
| nucleotide and amino acid sequences for cytochrome caa3-type oxidase of bacillus stearothermophilus k1041 and non-michaelis-type kinetics with cytochrome c. | a pseudo-sigmoidal cytochrome c-dependence curve of oxidase activity was observed with cytochrome oxidase from the bacillus stearothermophilus strain k1041, while the other thermophilic bacillus ps3 which has been extensively studied possessed normal michaelis-menten type kinetics. the genes coding for four subunits of cytochrome caa3-type oxidase and for heme o synthase were isolated from a genomic dna library of k1041 by using a ps3 dna fragment containing the highly-conserved region of the la ... | 1996 | 8611588 |
| cloning and sequencing of the dnak operon of bacillus stearothermophilus. | here, we report the cloning of a 5.8-kb psti fragment of chromosomal dna from bacillus stearothermophilus (bt) carrying the three genes, grpe, dnak and dnaj. this fragment contains, in addition, the 3'-end of an open reading frame which has been shown to be part of the dnak operon in three bacterial species. the dnaj gene could complement an escherichia coli dnajts mutant for growth at high temperature. sequencing and hybridization data strongly suggest that the bt chromosome contains an analog ... | 1996 | 8621094 |
| cyclization reaction catalyzed by branching enzyme. | the action of branching enzyme (ec 2.4.l.l8) from bacillus stearothermophilus on amylose was analyzed. the enzyme reduced the molecular size of amylose without increasing the reducing power. this result could not be explained by the normal branching reaction model. when the product was treated with glucoamylase (an exo++-type amylase), a resistant component remained. the glucoamylase-resistant component was easily digested by an endo-type alpha-amylase or by isoamylase plus glucoamylase. these r ... | 1996 | 8626287 |
| conversion from farnesyl diphosphate synthase to geranylgeranyl diphosphate synthase by random chemical mutagenesis. | prenyltransferases catalyze the consecutive condensation of isopentenyl diphosphate (ipp) with allylic diphosphates to produce prenyl diphosphates whose chain lengths are absolutely determined by each enzyme. in order to investigate the mechanisms of the consecutive reaction and of the determination of ultimate chain length, a random mutational approach was planned. the farnesyl diphosphate (fpp) synthase gene of bacillus stearothermophilus was subjected to random mutagenesis by nano2 treatment ... | 1996 | 8626566 |
| effectiveness of three types of sterilization on the contents of sharps containers. | the purpose of this study was to test the effect of treatment in a gravity steam autoclave, high-vacuum steam autoclave, or an unsaturated chemical vapor sterilizer on endospores present on strips or placed inside of dental anesthetic cartridges held within sharps containers. strips with 1.7 x 10(5) bacillus stearothermophilus endospores were used; the cartridges were soiled with an equal number of spores or spores mixed with blood. if sterilization was not accomplished after the initial period, ... | 1995 | 8628836 |
| m.bsshii, a multispecific cytosine-c5-dna-methyltransferase with unusual target recognizing properties. | a new multispecific cytosine-c5-dna-methyltransferase (c5-mtase), m.bsshii, was identified in bacillus stearothermophilus h3. the m.bsshii gene was cloned and sequenced. the amino acid sequence deduced shows the characteristic building plan of a c5-mtase. by sequencing bisulfite-treated dna methylated by m.bsshii and by restriction enzyme analysis, we defined the following methylation targets of m.bsshii: acgcgt/ccgcgg (mlui/sacii), pugcgcpy (haeii), puccggpy (cfr10i) and gcgcgc (bsshii). the re ... | 1996 | 8632477 |
| the lactose transporter in leuconostoc lactis is a new member of the lacs subfamily of galactoside-pentose-hexuronide translocators. | the gene encoding the lactose transport protein (lacs) of leuconostoc lactis nz6009 has been cloned from its native lactose plasmid, pnz63, by functional complementation of lactose permease-deficient escherichia coli mutants. nucleotide sequence analysis revealed an open reading frame with the capacity to encode a protein of 639 amino acids which had limited but significant identity to the lactose transport carriers (lacs) of streptococcus thermophilus (34.5%) and lactobacillus bulgaricus (35.6% ... | 1996 | 8633855 |
| phosphorus-31 nuclear magnetic resonance studies on coenzyme binding and specificity in glyceraldehyde-3-phosphate dehydrogenase. | binding of nad(p)+ to wild type and a series of mutants of the glycolytic nad-dependent glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus designed to alter the cofactor specificity [clermont, s., corbier, c., mely, y., gerard, d., wonacott, a., & branlant, g. (1993) biochemistry 21, 10178-10184] has been studied by 31p nmr. in the mutants with the l187a and p188s substitutions, the pyrophosphate signals are split, and the upfield resonance has been assigned to the ... | 1996 | 8634248 |
| purification and reconstitution of the glutamate carrier gltt of the thermophilic bacterium bacillus stearothermophilus. | an affinity tag consisting of six adjacent histidine residues followed by an enterokinase cleavage site was genetically engineered at the n-terminus of the glutamate transport protein gltt of the thermophilic bacterium bacillus stearothermophilus. the fusion protein was expressed in escherichia coli and shown to transport glutamate. the highest levels of expression were observed in e. coli strain dh5 alpha grown on rich medium. the protein could be purified in a single step by ni2+-nta affinity ... | 1996 | 8634258 |
| cooperative interactions of rna and thiostrepton antibiotic with two domains of ribosomal protein l11. | ribosomal protein l11 interacts with a 58-nucleotide domain of large subunit ribosomal rna; both the protein and its rna target have been highly conserved. the antibiotic thiostrepton recognizes the same rna domain, and binds to the ribosome cooperatively with l11. experiments presented here show that rna recognition and thiostrepton cooperativity can be attributed to c- and n-terminal domains of l11, respectively. under trypsin digestion conditions that degrade bacillus stearothermophilus l11 t ... | 1996 | 8634289 |
| sequence of the bacillus stearothermophilus gene encoding aspartokinase ii. | the complete nucleotide sequence of the gene encoding aspartokinase (ask) ii from thermophilic bacillus stearothermophilus has been determined. degenerate oligodeoxyribonucleotides primed the amplification of a 932-bp gene. this sequence was successively used for constructing new primers applied in inverse polymerase chain reaction using, as template, self-ligating dna fragments. the deduced amino-acid sequence is 68.7% identical with the sequence of the bacillus sp. strain mga3 ask ii. | 1996 | 8635739 |
| comparison of the structures of wild-type and a n313t mutant of escherichia coli glyceraldehyde 3-phosphate dehydrogenases: implication for nad binding and cooperativity. | the crystal structure of wild-type and n313t mutant glyceraldehyde 3-phosphate dehydrogenases from escherichia coli was determined in the presence of nad at 1.8 angstrom and 2.17 angstrom, respectively. the structure of the monomer and of the tetramer are similar to those observed for other gapdhs. an exhaustive analysis of the hydrophobic clusters and the hydrogen bond networks explain the high degree of sequence conservation in gapdhs. the structural effect of the n313t mutation is a change in ... | 1996 | 8636984 |
| structural basis for the extreme thermostability of d-glyceraldehyde-3-phosphate dehydrogenase from thermotoga maritima: analysis based on homology modelling. | d-glyceraldehyde-3-phosphate dehydrogenase (gapdh) from a hyperthermophilic eubacterium, thermotoga maritima, is remarkably heat stable (tm = 109 degrees c). in this work, we have applied homology modelling to predict the 3-d structure of th.maritima gapdh to reveal the structural basis of thermostability. three known gapdh structures were used as reference proteins. first, the rough model of one subunit was constructed using the identified structurally conserved and variable regions of the refe ... | 1995 | 8637847 |
| late events in translation initiation. adjustment of fmet-trna in the ribosomal p-site. | the requirements for the adjustment of fmet-trna in the ribosomal p-site have been analyzed by studying the formation of fmet-puromycin in a bacillus stearothermophilus system. the binding of fmet-trna to the 30 s ribosomal subunit is not drastically affected by the omission of gtp, mrna, mrna and gtp, or by replacing gtp with gtp analogues. the adjustment of fmet-trna in the p site has stricter requirements and fmet-puromycin formation occurred at its maximum rate and extent when fmet-trna was ... | 1996 | 8642589 |
| protein recognition of a ribosomal rna tertiary structure. | ribosomal protein l11 recognizes a highly conserved, 58 nucleotide domain of large subunit ribosomal rna. this domain has a set of tertiary interactions that are specifically stabilized by mg2+ and nh4+ ions. the protein recognizes this tertiary structure, in the sense that ions stabilizing the tertiary structure also promote l11 binding, and a heat stable form of the protein (from bacillus stearothermophilus) prevents rna unfolding. we also find that these rna-binding properties are confined to ... | 1995 | 8643396 |
| bacillus stearothermophilus qcr operon encoding rieske fes protein, cytochrome b6, and a novel-type cytochrome c1 of quinol-cytochrome c reductase. | the gcr of bacillus stearothermophilus k1041 encoding three subunits of the quinol-cytochrome c oxidoreductase (cytochrome reductase, b6c1 complex) was cloned and sequenced. the gene (qcra) for a rieske fes protein of 19,144 da with 169 amino acid residues, and the gene (qcrc) for cytochrome c1 of 27,342 da with 250 amino acid residues were found at adjacent upstream and downstream sides of the previously reported qcrb (petb) for cytochrome b6 of subunit 25,425 da with 224 residues (sone, n., sa ... | 1996 | 8647852 |
| effect on ph heating medium on the thermal resistance of bacillus stearothermophilus spores. | the influence of the ph of heating medium on heat resistance of bacillus stearothermophilus spores (atcc 7953, 12980, 15951 and 15952) were studied. the ph values tested were: 4.0, 5.0, 6.0 and 7.0 at temperatures of 115, 120, 125, 130 and 135 degrees c. it was found that at low treatment temperatures (115 degrees c) d-values decreased between 7- and 10-fold with 7953, 12980 and 15951 strains and about 23-fold with 15952 strain when ph dropped from 7.0 to 4.0. at highest treatment temperatures ( ... | 1996 | 8652348 |
| phosphate-binding sites in phosphorylating glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | the two anion-binding sites of the glycolytic glyceraldehyde-3-phosphate dehydrogenase (grap-dh), the ps and pi sites, were originally proposed by moras et al. [moras, d., olsen, k.w., sabesan, m.n., buehner, m., ford, g.c. & rossmann, m. g. (1975) j. biol. chem. 250, 9137-9162] to bind the c3 phosphate of the glyceraldehyde 3-phosphate and the inorganic phosphate respectively. ps site mutants t179a, and t179m, and r231l, and the pi site mutants t150a and t208 of the bacillus stearothermophilus ... | 1996 | 8654412 |
| bstf5i, an unusual isoschizomer of foki. | bstf5i, a new restriction endonuclease (enase) from bacillus stearothermophilus f5, has been discovered. this enzyme recognizes 5'-ggatg-3' and cleaves dna, generating a 2-base 3'extension: 5'-ggatg nn[symbol: see text]-3' 3'-cctac[symbol: see text]nn-5' bstf5i is an isoschizomer of foki and seems to be evolutionarily close to other nonpalindromic-recognizing enases from thermophilic bacilli. | 1996 | 8654990 |
| the caa3 terminal oxidase of bacillus stearothermophilus. transient spectroscopy of electron transfer and ligand binding. | the thermophilic bacterium bacillus stearothermophilus possesses a caa3-type terminal oxidase, which was previously purified (de vrij, w., heyne, r. i. r., and konings, w. n. (1989) eur. j. biochem. 178, 763-770). we have carried out extensive kinetic experiments on the purified enzyme by stopped-flow time-resolved optical spectroscopy combined with singular value decomposition analysis. the results indicate a striking similarity of behavior between this enzyme and the electrostatic complex betw ... | 1996 | 8662862 |
| structure of the r65q mutant of yeast 3-phosphoglycerate kinase complexed with mg-amp-pnp and 3-phospho-d-glycerate. | the structure of a ternary complex of the r65q mutant of yeast 3-phosphoglycerate kinase (pgk) with magnesium 5'-adenylylimidodiphosphate (mg-amp-pnp) and 3-phospho-d-glycerate (3-pg) has been determined by x-ray crystallography to 2.4 angstrom resolution. the structure was solved by single isomorphous replacement, anamalous scattering, and solvent flattening and has been refined to an r-factor of 0.185, with rms deviations from ideal bond distance and angles of 0.009 angstrom and 1.78 degrees, ... | 1996 | 8672447 |
| construction of single amino acid substitution mutants of cloned bacillus stearothermophilus dna polymerase i which lack 5'-->3' exonuclease activity. | two individual amino acid substitutions were engineered at a selected site in the 5' --> 3' exonuclease domain of the cloned bacillus stearothermophilus dna polymerase i gene. these mutations resulted in the expression of enzymes lacking the 5' --> 3' exonuclease activity while maintaining normal polymerizing activity. the mutated and non-mutated enzymes were each constitutively expressed in an escherichia coli host without the use of an exogenous or inducible promoter, and the mutated enzymes w ... | 1996 | 8679703 |
| structural consequences of neopullulanase mutations. | bacillus stearothermophilus neopullulanase (npl) structure was modeled based on aspergillus oryzae alpha-amylase (taa) to understand the structure-function relationships of this pullulan hydrolyzing enzyme. the npl structure seems to consist of a central (alpha/beta)8 barrel to which the other domains are attached. the immediate surroundings of the npl catalytic site were found to have very similar structure to taa. the more distant sites are different due to the stereochemical requirements of a ... | 1996 | 8695646 |
| microwave continuous sterilization of injection ampoules. | a new microwave continuous sterilizer (mws) for applying microwave dielectric heating as an alternative to an autoclave was developed. the developmental objectives of the mws were: 1. achieving sufficient sterilization for the drugs containing heat-sensitive ingredients. 2. measuring and recording sterilization temperature of each ampoule. 3. ensuring automatic continuous operation and linkage with the preceding and following machines in an injection ampoule production process. the temperature o ... | 1996 | 8696781 |
| purification and characterization of a malic enzyme from the ruminal bacterium streptococcus bovis atcc 15352 and cloning and sequencing of its gene. | malic enzyme (ec 1.1.1.39), which catalyzes l-malate oxidative decarboxylation and pyruvate reductive carboxylation, was purified to homogeneity from streptococcus bovis atcc 15352, and properties of this enzyme were determined. the 2.9-kb fragment containing the malic enzyme gene was cloned, and the sequence was determined and analyzed. the enzymatic properties of the s. bovis malic enzyme were almost identical to those of other malic enzymes previously reported. however, we found that the s. b ... | 1996 | 8702261 |
| cloning, sequencing and functional overexpression of the streptococcus equisimilis h46a gapc gene encoding a glyceraldehyde-3-phosphate dehydrogenase that also functions as a plasmin(ogen)-binding protein. purification and biochemical characterization of the protein. | we previously identified dna sequences involved in the function of the complex promoter of the streptokinase gene from streptococcus equisimilis h46a, a human serogroup c strain known to express this gene at a high level. as a prerequisite to understanding possible mechanisms that control the balance between the plasminogen activating and plasmin(ogen) binding capacities of h46a, we describe here its gapc gene encoding glyceraldehyde-3-phosphate dehydrogenase (grap-dh, ec 1.2.1.12), a glycolytic ... | 1996 | 8706717 |
| evaluation of injections of collagenase and oxytetracycline via the umbilical artery as treatment for retained placenta in cattle. | to test whether oxytetracycline inactivates collagenase when combined as a potential treatment for retained fetal membranes in cattle and to determine whether oxytetracycline passes to blood from fetal membranes after intraplacental injection. | 1996 | 8712518 |
| structures of prokaryotic ribosomal proteins: implications for rna binding and evolution. | after a long hiatus, the pace of determination of the structures of ribosomal proteins has accelerated dramatically. we discuss here the structures of five ribosomal proteins from bacillus stearothermophilus: s5, s17, l6, l9, and l14. these structures represent several new motifs. each of these structures has revealed new insights, and we have developed criteria for recognizing rna-binding regions of each protein and correlating the structures with such properties as antibiotic resistance. the i ... | 1995 | 8722013 |
| translational regulation of infc operon in bacillus stearothermophilus. | a bacillus stearothermophilus in vitro translational system has been developed to study the expression of the three cistrons (infc, rpml, and rplt) constituting the infc operon of this bacterium. when directed by homologous in vitro transcribed infc tricistronic mrna, this system, which consists of partially purified and purified components of the b. stearothermophilus translational apparatus, synthesizes with high efficiency and specificity the three gene products (if3, l35, and l20) in a ratio ... | 1995 | 8722023 |
| regulation of the escherichia coli s10 ribosomal protein operon by heterologous l4 ribosomal proteins. | we have cloned the l4 ribosomal protein genes from morganella morganii and haemophilus influenza. the sequences of these genes were compared with published sequences for escherichia coli, yersinia pseudotuberculosis, and bacillus stearothermophilus. all five of these l4 genes were expressed in e. coli and shown to function as repressors of both transcription and translation of the e. coli s10 operon. possible implications for regulation of r-protein synthesis in species other e. coli are discuss ... | 1995 | 8722027 |
| structural and functional implications in the eubacterial ribosome as revealed by protein-rrna and antibiotic contact sites. | contact sites between protein and rrna in 30s and 50s ribosomal subunits of escherichia coli and bacillus stearothermophilus were investigated at the molecular level using uv and 2-iminothiolane as cross-linkers. thirteen ribosomal proteins (s3, s4, s7, s14, s17, l2, l4, l6, l14, l27, l28, l29, and l36) from these organisms were cross-linked in direct contact with the rnas, and the peptide stretches as well as amino acids involved were identified. further, the binding sites of puromycin and spir ... | 1995 | 8722036 |
| purification and characterization of the alcohol dehydrogenase from a novel strain of bacillus stearothermophilus growing at 70 degrees c. | the biocatalysts isolated from thermophilic microorganisms are the object of ever-growing scientific interest for (i) the comprehension of the molecular basis of their thermal tolerance, and (ii) their use in different bio-industrial fields. here we report the purification and characterization of an alcohol dehydrogenase (designated adh-ht) from the novel strain lld-r of bacillus stearothermophilus which grows at 70 degrees c. adh-ht was obtained in pure form by anion exchange chromatography and ... | 1996 | 8729010 |
| thermostable bst dna polymerase i lacks a 3'-->5' proofreading exonuclease activity. | a thermostable dna polymerase, the bst dna polymerase i, from bacillus stearothermophilus n3468 was prepared to near-homogeneity. the dominant species of the bst dna polymerase i preparation sized about 97 kda when analyzed on sds polyacrylamide gels. the bst pola gene that codes for bst polymerase i was cloned and sequenced. comparative sequence analysis showed that all three conserved 3'-->5' exonuclease motifs found in e. coli dna polymerase i were missing in bst dna polymerase i. this cast d ... | 1996 | 8740835 |
| lysyl-trna synthetase from bacillus stearothermophilus. purification, and fluorometric and kinetic analysis of the binding of substrates, l-lysine and atp. | lysyl-trna synthetase [l-lysine:trna(lys)ligase (amp forming); ec 6.1.1.6] was purified from bacillus stearothermophilus nca1503 approximately 1,100-fold to homogeneity in page. the enzyme is a homodimer of m(r) 57,700 x 2. the molar absorption coefficient, epsilon, at 280 nm is 71,600 m-1.cm-1 at ph8.0. enzyme activity in the trna aminoacylation reaction and the atp-ppi exchange reaction increases up to 50 degrees c at ph 8.0, but is lost completely at 70 degrees c. the ph-optima of the two rea ... | 1996 | 8743569 |
| identification of significant residues in the substrate binding site of bacillus stearothermophilus farnesyl diphosphate synthase. | farnesyl diphosphate synthases have been shown to possess seven highly conserved regions (i-vii) in their amino acid sequences [koyama et al. (1993) j. biochem. (tokyo) 113, 355-363]. site-directed mutants of farnesyl diphosphate synthase from bacillus stearothermophilus were made to evaluate the roles of the conserved aspartic acids in region vi and lysines in regions i, v, and vi. the aspartate at position 224 was changed to alanine or glutamate (mutants designated as d224a and d224e, respecti ... | 1996 | 8755734 |
| the bacillus stearothermophilus nub36 sura gene encodes a thermophilic sucrase related to bacillus subtilis saca. | the complete nucleotide sequence of the sura gene, encoding a sucrase from bacillus stearothermophilus nub36, was determined. sura was composed of 1338 bp and encoded 445 amino acid residues. the deduced polypeptide of m(r) 51519 showed strong sequence similarity to sucrose and sucrose phosphate hydrolases from bacillus subtilis, klebsiella pneumoniae and vibrio alginolyticus, and contained the 'sucrose box' residues thought to be important for catalysis of the transfer of fructose from sucrose. ... | 1996 | 8757729 |
| a concerted tryptophanyl-adenylate-dependent conformational change in bacillus subtilis tryptophanyl-trna synthetase revealed by the fluorescence of trp92. | a semi-conserved tryptophan residue of bacillus subtilis tryptophanyl-trna synthetase (trprs) was previously asserted to be an essential residue and directly involved in trnatrp binding and recognition. the crystal structure of the bacillus stearothermophilus trprs tryptophanyl-5'-adenylate complex (trp-amp) shows that the corresponding trp91 is buried and in the dimer interface, contrary to the expectations of the earlier assertation. here we examine the role of this semi-conserved tryptophan r ... | 1996 | 8757806 |
| microbiologic assay for detection of antimicrobial agents in urine. | antimicrobial therapy can be a confounding factor in the diagnosis of urinary tract infection and is not always reported to laboratories by physicians. we developed a microbiologic assay for screening urine specimens for antimicrobial agents. bacillus stearothermophilus was used as the indicator bacteria. a total of 1,921 urine specimens from three hospitals in taiwan were screened using this assay. of the samples assayed, 1,293 were positive for antimicrobial agents. agreement between informati ... | 1996 | 8772053 |
| solution structure of the lipoyl domain of the 2-oxoglutarate dehydrogenase complex from azotobacter vinelandii. | the three-dimensional solution structure of the lipoyl domain of the 2-oxoglutarate dehydrogenase complex from azotobacter vinelandii has been determined from nuclear magnetic resonance data by using distance geometry and dynamical simulated annealing refinement. the structure determination is based on a total of 580 experimentally derived distance constraints and 65 dihedral angle constraints. the solution structure is represented by an ensemble of 25 structures with an average root-mean-square ... | 1996 | 8780784 |
| purification and characterization of purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase from bacillus stearothermophilus th 6-2. | the purine nucleoside phosphorylase (pu-npase) and the pyrimidine nucleoside phosphorylase (py-npase) have been purified from bacillus stearothermophilus th 6-2. the pu-npase is a trimer of 30-kda subunits and the py-npase is a dimer of 46-kda subunits. the isoelectric points of pu-npase and py-npase were ph 4.3 and 4.6, respectively. the pu-npase could catalyze the phosphorolysis of inosine and guanosine, but not adenosine. the py-npase could phosphorolyze both uridine and thymidine. | 1996 | 8782414 |
| interaction of the bacillus stearothermophilus ribosomal protein s15 with 16 s rrna: i. defining the minimal rna site. | the ribosomal rna binding site of bacillus stearothermophilus ribosomal protein s15 (bs15) was analyzed using synthetic rna oligonucleotides derived from the 16 s rrna central domain. native gel electrophoresis mobility shift assays demonstrate that bs15 can specifically interact with an rna oligonucleotide containing nucleotides 585 to 756 (helices 20 to 23) of 16 s rrna with an apparent dissociation constant of 35 nm. a series of deletion mutants of the rrna fragment that contains the bs15 spe ... | 1996 | 8794875 |
| interaction of the bacillus stearothermophilus ribosomal protein s15 with 16 s rrna: ii. specificity determinants of rna-protein recognition. | s15 is a primary ribosomal protein that interacts specifically with a three-way junction in the central domain of 16 s rrna, whose binding induces a conformational change in the rna. in the accompanying paper, we demonstrated that s15 binds with high affinity to a 61 nucleotide rna corresponding to the minimal rrna binding site. here, the sequence and structural determinants for the rna in the bacillus stearothermophilus s15-rrna interaction have been probed using site-directed mutagenesis, chem ... | 1996 | 8794876 |
| cumulative stabilizing effects of hydrophobic interactions on the surface of the neutral protease from bacillus subtilis. | using genetically engineered mutants of the neutral protease from bacillus stearothermophilus (bstenp), it had been shown that the surface-exposed structural motif constituted by phe63 embedded in a four amino acid hydrophobic pocket is critical for the thermal stability of the thermophilic neutral proteases from bacilli. to measure the stabilizing contribution of each hydrophobic interaction taking place between phe63 and the hydrophobic pocket, we grafted this structural motif in the neutral p ... | 1996 | 8795044 |
| heat resistance of bacillus stearothermophilus spores in alginate-mushroom puree mixture. | thermal resistance of bacillus stearothermophilus spores has been established inoculating spores in alginate-mushroom puree mixture (ungelled) and in alginate-mushroom puree mixture set in calcium chloride (gelled). data are compared with those obtained suspending the spores in distilled water, mushroom extract and in calcium chloride. results indicated that, in general, d values obtained in gelled mixture were higher than d values obtained in distilled water, mushroom extract or in ungelled mix ... | 1996 | 8796439 |
| secretion of bacillus alpha-amylase from yeast directed by glucoamylase i signal sequence of saccharomyces diastaticus. | for the secretion of bacillus stearothermophilus alpha-amylase from yeast, a recombinant plasmid pgat17 was constructed by fusing b. stearothermophilus alpha-amylase structural gene in frame to the promoter and signal sequence of saccharomyces diastaticus glucoamylase i gene (sta1). the secretion of the heterologous alpha-amylase from s. diastaticus transformed with pgat17 was confirmed by the halo formation around colonies on selective starch agar medium. about 80% of the total alpha-amylase ac ... | 1996 | 8799340 |
| increase of cytochrome b mrna by bis(2-hydroxyethyl) trisulfide in j774a.1 cells. | the effect of a cytotoxic substance (bis(2-hydroxyethyl) trisulfide; bs-1), isolated from bacillus stearothermophilus uk563, on messenger ribonucleic acid (mrna) level in mouse macrophage-like j774a.1 cells was investigated by the method of differential display. the treatment of j774a.1 cells with bs-1 led us to detect the inducible gene. the sequence analysis revealed that the gene was identical to mouse mitochondrial cytochrome b. in fact, northern blot analysis showed that cytochrome b mrna i ... | 1996 | 8799491 |
| the arginine operon of bacillus stearothermophilus: characterization of the control region and its interaction with the heterologous b. subtilis arginine repressor. | mechanisms of gene regulation have not yet been extensively studied in thermophilic bacteria. in previous studies we showed that the bacillus stearothermophilus argcjbd gene cluster is subject to specific repression by arginine. here we report the cloning by colony hybridization, and characterization of the proximal part of the argc gene together with the adjacent control region of the cluster. the promoter was identified by primer extension mapping of the argc transcription startpoint: a sequen ... | 1996 | 8804405 |
| the crystal structure of ribosomal protein l14 reveals an important organizational component of the translational apparatus. | detailed structural information on ribosomal proteins has increased our understanding of the structure, function and evolution of the ribosome. l14 is one of the most conserved ribosomal proteins and appears to have a central role in the ribonucleoprotein complex. studies have indicated that l14 occupies a central location between the peptidyl transferase and gtpase regions of the large ribosomal subunit. | 1996 | 8805509 |
| protein-protein interactions in the pyruvate dehydrogenase multienzyme complex: dihydrolipoamide dehydrogenase complexed with the binding domain of dihydrolipoamide acetyltransferase. | the ubiquitous pyruvate dehydrogenase multienzyme complex is built around an octahedral or icosahedral core of dihydrolipoamide acetyltransferase (e2) chains, to which multiple copies of pyruvate decarboxylase (e1) and dihydrolipoamide dehydrogenase (e3) bind tightly but non-covalently. e2 is a flexible multidomain protein that mediates interactions with e1 and e3 through a remarkably small binding domain (e2bd). | 1996 | 8805537 |
| structural evidence for specific s8-rna and s8-protein interactions within the 30s ribosomal subunit: ribosomal protein s8 from bacillus stearothermophilus at 1.9 a resolution. | prokaryotic ribosomal protein s8 is an important rna-binding protein that occupies a central position within the small ribosomal subunit. it interacts extensively with 16s rrna and is crucial for the correct folding of the central domain of the rrna. s8 also controls the synthesis of several ribosomal proteins by binding to mrna. it binds specifically to very similar sites in the two rna molecules. | 1996 | 8805594 |
| thermal resistance of bacillus stearothermophilus spores on strips previously treated with calcium. | moist thermal resistance parameters of spores of b. stearothermophilus atcc 7953, inoculated in aqueous suspensions and on strips, were evaluated at 118 degrees c and 121 degrees c, by the proposed serum-bottle-mini-autoclave method. the present work studied the effect of alkaline calcium exchange-inducing storage treatments on the viability and moist heat-resistance of b. stearothermophilus spores, stored at -18 degrees c, following impregnation into filter paper strips which had been previousl ... | 1996 | 8810838 |
| effects of nad+ binding on the luminescence of tryptophans 84 and 310 of glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | the individual fluorescence and phosphorescence properties of w84 and w310 in bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase were identified through the construction of a single tryptophan mutant (w84f) and by comparison of the emission between mutant and wild-type enzymes. the results show that the luminescence of w310 is red-shifted and substantially quenched relative to that of w84. it displays an average subnanosecond fluorescence lifetime (tau f) and a very short, 50 m ... | 1996 | 8823192 |
| cloning, expression, and isolation of the mannitol transport protein from the thermophilic bacterium bacillus stearothermophilus. | a mannitol phosphotransferase system (pts) was identified in bacillus stearothermophilus by in vitro complementation with escherichia coli ei, hpr, and iia(mtl). degenerate primers based on regions of high amino acid similarity in the e. coli and staphylococcus carnosus eii(mt1) were used to develop a digoxigenin-labeled probe by pcr. using this probe, we isolated three overlapping dna fragments totaling 7.2 kb which contain the genes mtla, mtlr, mtlf, and mtld, encoding the mannitol iicb,a regu ... | 1996 | 8824601 |
| characterization and expression of the plasmid-borne bedd gene from pseudomonas putida ml2, which codes for a nad+-dependent cis-benzene dihydrodiol dehydrogenase. | the catabolic plasmid phmt112 in pseudomonas putida ml2 contains the bed gene cluster encoding benzene dioxygenase (bedc1c2ba) and a nad+-dependent dehydrogenase (bedd) required to convert benzene into catechol. analysis of the nucleotide sequence upstream of the benzene dioxygenase gene cluster (bedc1c2ba) revealed a 1,098-bp open reading frame (bedd) flanked by two 42-bp direct repeats, each containing a 14-bp sequence identical to the inverted repeat of is26. in vitro translation analysis sho ... | 1996 | 8824602 |
| evidence that an n-terminal s-layer protein fragment triggers the release of a cell-associated high-molecular-weight amylase in bacillus stearothermophilus atcc 12980. | during growth on starch medium, the s-layer-carrying bacillus stearothermophilus atcc 12980 and an s-layer-deficient variant each secreted three amylases, with identical molecular weights of 58,000, 122,000, and 184,000, into the culture fluid. only the high-molecular-weight amylase (hmwa) was also identified as cell associated. extraction and reassociation experiments showed that the hmwa had a high-level affinity to the peptidoglycan-containing layer and to the s-layer surface, but the interac ... | 1996 | 8824603 |
| heterologous expression and self-assembly of the s-layer protein sbsa of bacillus stearothermophilus in escherichia coli. | the cell surface of bacillus stearothermophilus pv72 is covered by a regular surface layer (s-layer) composed of single species of protein, sbsa, with a molecular weight of 130,000. recently, the sequence of the corresponding gene (sbsa) has been determined. the sbsa coding region including the signal sequence was cloned as a polymerase chain reaction (pcr) product into a low-copy-number vector under the transcriptional control of the lambda pl promoter. expression of sbsa was shown to be therma ... | 1996 | 8830240 |
| comparison of ion plasma, vaporized hydrogen peroxide, and 100% ethylene oxide sterilizers to the 12/88 ethylene oxide gas sterilizer. | the performance of a standard gas sterilizer, which uses a mixture of 12% ethylene oxide (eto) and 88% chlorofluorocarbon as the sterilizing gas (12/88), was compared to selected gas, ion plasma, and vaporized hydrogen peroxide (h2o2) sterilizers that do not use chlorofluorocarbons. the effect of serum and salt on sterilizer performance was evaluated. | 1996 | 8835444 |
| stereospecific production of the herbicide phosphinothricin (glufosinate): purification of aspartate transaminase from bacillus stearothermophilus, cloning of the corresponding gene, aspc, and application in a coupled transaminase process. | we have isolated and characterized an aspartate transaminase (glutamate:oxalacetate transaminase, ec 2.6.1.1) from the thermophilic microorganism bacillus stearothermophilus. the purified enzyme has a molecular mass of 40.5 kda by sodium dodecyl sulfate gel analysis, a temperature optimum of 95 degrees c, and a ph optimum of 8.0. the corresponding gene, aspc, was cloned and overexpressed in escherichia coli. the recombinant glutamate:oxalacetate transaminase protein was used in immobilized form ... | 1996 | 8837436 |
| cytochrome bd-type quinol oxidase in a mutant of bacillus stearothermophilus deficient in caa3-type cytochrome c oxidase. | gram-positive thermophilic bacillus species contain cytochrome caa3-type cytochrome c oxidase as a terminal oxidase in the respiratory chain. to identify alternative oxidases, we isolated b. stearothermophilus mutants defective in the caa3-type oxidase activity. one mutant contained little cytochrome a and had low cytochrome c oxidase activity. however, growth and the respiratory activity of membranes in the presence of nadh were close to normal, suggesting that the mutant contains an alternativ ... | 1996 | 8837467 |
| discussion of the z-value to use in calculating the f0-value for high-temperature sterilization processes. | the appropriate z-value to use in integrating heat process time-temperature data in the temperature range of 120.0-140.0 degrees c (248.0-284.0 degrees f) is discussed. we conclude that for control of clostridium botulinum there is little risk in extrapolating a public health f0-value for c. botulinum to temperatures in the 132.0-138.0 degrees c (270.0-280.0 degrees f) range using a z-value of 10.0 degrees c (18.0 degrees f). it would seem prudent, at this time, when extrapolating data to condit ... | 1996 | 8846059 |
| biological indicators for low temperature steam and formaldehyde sterilization: investigation of the effect of change in temperature and formaldehyde concentration on spores of bacillus stearothermophilus ncimb 8224. | five strains of bacillus stearothermophilus have been studied to identify a spore strain to be used as a biological indicator organism for low temperature steam and formaldehyde sterilization. three strains gave poor reproducibility of batch size and growth index and were discarded. the other two strains gave good reproducibility with a high growth index and gave rise to linear survivor curves when exposed to 5% aqueous formaldehyde. however, only ncimb 8224 sporulates on a simpler medium and as ... | 1996 | 8852673 |
| inactivation of bacteria and yeasts on agar surfaces with high power nd:yag laser light. | near infrared light from a high-powered, 1064 nm, neodymium:yttrium aluminium garnet (nd:yag) laser killed a variety of gram-positive and gram-negative bacteria and two yeasts, lawned on nutrient agar plates. a beam (cross-sectional area, 1.65 cm2) of laser light was delivered in 10 j, 8 ms pulses at 10 hz, in a series of exposure times. for each microbial species, a dose/response curve was obtained of area of inactivation vs energy density (j cm-2). the energy density that gave an inactivation ... | 1996 | 8862017 |
| thermal resistance of bacillus stearothermophilus spores in different heating systems containing some approved food additives. | the effects of different heating systems on the heat resistance of bacillus stearothermophilus spores (atcc 7953, 12980, 15951 and 15952) were investigated. spores were heated in distilled water, sorensen buffer (0.18 mol 1-1), mcilvaine buffer (0.0025-0.18 mol 1-1), and several solutions containing sodium chloride (0.06-12%), sodium nitrite (125 ppm), potassium sorbate (0.1%) and sodium benzoate (0.1%) over a wide range of temperatures (115-140 degrees c). d-values obtained for mcilvaine and so ... | 1996 | 8862025 |
| detection of m. tuberculosis dna using thermophilic strand displacement amplification. | strand displacement amplification (sda) is an isothermal, in vitro method of amplifying dna that is based upon the combined action of a dna polymerase and restriction enzyme. previously, a form of sda was developed which utilizes the exonuclease deficient klenow fragment of e. coli polymerase i (exo klenow) and the restriction enzyme hincii to achieve 10(8)-fold amplification in 2 h at 37 degrees c (walker, g.t., 1993, pcr methods and applications 3; 1-6). a new thermophilic form of sda is repor ... | 1996 | 8865173 |
| use of graph theory for secondary structure recognition and sequential assignment in heteronuclear (13c, 15n) nmr spectra: application to hu protein from bacillus stearothermophilus. | a computer-assisted procedure, based upon a branch of mathematics known as graph theory, has been developed to recognize secondary structure elements in proteins from their corresponding nuclear overhauser effect spectroscopy (noesy)-type spectra and to carry out their sequential assignment. in the method, noe connectivity templates characteristic of regular secondary structures are identified in the spectra. resonance assignment is then achieved by connecting these noe patterns of secondary str ... | 1996 | 8875823 |
| properties of diacetyl (acetoin) reductase from bacillus stearothermophilus. | the cells of bacillus stearothermophilus contain an nadh-dependent diacetyl (acetoin) reductase. the enzyme was easily purified to homogeneity, partially characterised, and found to be composed of two subunits with the same molecular weight. in the presence of nadh, it catalyses the stereospecific reduction of diacetyl first to (3s)-acetoin and then to (2s,3s)-butanediol; in the presence of nad+, it catalyses the oxidation of (2s,3s)- and meso-butanediol, respectively, to (3s)-acetoin and to (3r ... | 1996 | 8879540 |
| mathematical model for the combined effect of temperature and ph on the thermal resistance of bacillus stearothermophilus and clostridium sporogenes spores. | two mathematical models have been studied to establish the relationship between the ph, treatment temperature and thermal destruction constant (k) of bacillus stearothermophilus and clostridium sporogenes spores. the study was carried out by heating the spores in mushroom extract acidified with two different acidulants (citric acid and glucono-delta-lactone). among the models studied, the one that best described the inactivation was a second order polynomial equation, the precision of which depe ... | 1996 | 8880342 |
| functional characterization of an extreme thermophilic class ii fructose-1,6-bisphosphate aldolase. | fructose-1,6-bisphosphate aldolase activity has been isolated and purified to homogeneity from the extreme thermophile eubacteria thermus aquaticus. the homogeneous enzyme is a class ii aldolase as fructose-1,6-bisphosphate cleavage activity was strongly inhibited by edta, and activated by co2+ metal ion. taq aldolase is a stable tetramer with estimated molecular mass of 165 kda. the enzyme is thermostable and is not inactived after heating at 90 degrees c for 2 h but looses 80% of activity afte ... | 1996 | 8898912 |
| refolding and reassociation of glycerol dehydrogenase from bacillus stearothermophilus in the absence and presence of groel. | the refolding of the tetrameric, metalloenzyme glycerol dehydrogenase (gdh) from bacillus stearothermophilus has been investigated using stopped-flow fluorescence and circular dichroism spectroscopy. the effects of metal ions on the refolding of the native enzyme and the refolding of a monomeric mutant ([a208]gdh) have also been studied. the refolding process of the wild-type enzyme is at least biphasic; 70% of the respective signal changes occur in the first 2 ms followed by a slower process wi ... | 1996 | 8917453 |
| recognition of a surface loop of the lipoyl domain underlies substrate channelling in the pyruvate dehydrogenase multienzyme complex. | in the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus, the interaction between the pyruvate decarboxylase (e1p) component and the lipoyl domain of the dihydrolipoyl acetyltransferase (e2) component was investigated using a combination of site-directed mutagenesis and nmr spectroscopy. residues 11 to 15 (egihe) of the lipoyl domain, part of a surface loop close in space to the beta-turn containing the lipoyl-lysine residue (position 42), were deleted or replaced. the mu ... | 1996 | 8918601 |
| crystal structure of a dexx box dna helicase. | there are a wide variety of helicases that unwind helical dna and rna substrates. the twelve helicases that have been identified in escherichia coli play a role in almost all cellular processes involving nucleic acids. we have solved the crystal structure of a monomeric form of a dna helicase from bacillus stearothermophilus, alone and in a complex with adp, at 2.5 and 2.9 a resolution, respectively. the enzyme comprises two domains with a deep cleft running between them. the atp-binding site, w ... | 1996 | 8934527 |
| the sterilization of endodontic hand files. | several different methods of file sterilization were analyzed to determine the best method of providing complete file sterility, including the metal shaft and plastic handle. six test groups of 15 files were studied using bacillus stearothermophilus as the test organism. groups were "sterilized" by glutaraldehyde immersion, steam autoclaving, and various techniques of salt sterilization. only proper steam autoclaving reliably produced completely sterile instruments. salt sterilization and glutar ... | 1996 | 8934994 |
| a role of the amino acid residue located on the fifth position before the first aspartate-rich motif of farnesyl diphosphate synthase on determination of the final product. | farnesyl diphosphate (fpp) synthase catalyzes consecutive condensations of isopentenyl diphosphate with allylic substrates to give fpp, c-15 compound, as a final product and does not catalyze a condensation beyond fpp. recently, it was observed that, in bacillus stearothermophilus fpp synthase, a replacement of tyrosine with histidine at position 81, which is located on the fifth amino acid before the first aspartate-rich motif, caused the mutated fpp synthase to catalyze geranylgeranyl diphosph ... | 1996 | 8940054 |