| a single amino acid substitution in lactate dehydrogenase improves the catalytic efficiency with an alternative coenzyme. | using site-directed mutagenesis, the nadh-linked lactate dehydrogenase from bacillus stearothermophilus has been specifically altered at a single residue to shift the coenzyme specificity towards nadph. the single change is at position 53 in the amino acid sequence where a conserved aspartate has been replaced by a serine. this substitution was made to reduce steric hindrance on binding of the extra phosphate group of nadph and to remove the negative charge of the aspartate group. the resultant ... | 1990 | 2302233 |
| a new type ii restriction endonuclease, bsma i, from bacillus stearothermophilus. | | 1990 | 2308865 |
| colorimetric glucose assay using thermostable glucokinase. | a method for assaying glucose in serum or plasma samples using a thermostable glucokinase was developed. glucokinase from bacillus stearothermophilus was coupled with glucose-6-phosphate dehydrogenase to produce nadph, which reduced the tetrazolium dye mtt to its formazan. detection of the product at 660 nm allowed samples containing up to 30 mmol/l glucose to be assayed with an endpoint method. use of the optimal wavelength for formazan detection, 570 nm, increased sensitivity for nadph detecti ... | 1990 | 2310155 |
| direct phase determination for the molecular envelope of tryptophanyl-trna synthetase from bacillus stearothermophilus by x-ray contrast variation. | monoclinic crystals of bacillus stearothermophilus tryptophanyl-trna synthetase grown in the presence of substrate tryptophan (space group p2(1)) display evidence of a low-resolution trigonal space group (p321). the origin and averaging transformations for the local 32 point group of this unusually clear sixfold non-crystallographic symmetry may be inferred without prior estimation of the electron density. this local symmetry was exploited in conjunction with solvent density contrast variation t ... | 1990 | 2310535 |
| electron transferase activity of diaphorase (nadh: acceptor oxidoreductase) from bacillus stearothermophilus. | electrochemical kinetic measurements were carried out for electron-transfer between nadh and the oxidized forms of mediators (ferrocenylmethanol (fma), ferrocenyl-1-ethanol (fea), n,n,n',n'-tetramethylphenylenediamine (tmpd), co(phen)2+(3) and fe(cn)4-(6)) catalyzed by diaphorase (nadh: acceptor oxidoreductase, ec 1.6.99.-) purified from bacillus stearothermophilus. cyclic voltammograms for the mediators with excess nadh in the presence of diaphorase gave steady-state currents. the quantitative ... | 1990 | 2317516 |
| random mutagenesis used to probe the structure and function of bacillus stearothermophilus alpha-amylase. | mutations that cover the sequence of bacillus stearothermophilus alpha-amylase were produced by an efficient in vitro enzymatic random mutagenesis method and the mutant alpha-amylases were expressed in escherichia coli, which also secreted the product. ninety-eight mutants were identified by sequencing and their enzyme activities were classified into three classes: wild-type, reduced or null. a molecular model of the enzyme was constructed using the coordinates of takaamylase a and a consensus a ... | 1990 | 2330367 |
| structure determination and refinement of bacillus stearothermophilus lactate dehydrogenase. | structures have been determined of bacillus stearothermophilus "apo" and holo lactate dehydrogenase. the holo-enzyme had been co-crystallized with the activator fructose 1,6-bisphosphate. the "apo" lactate dehydrogenase structure was solved by use of the known apo-m4 dogfish lactate dehydrogenase molecule as a starting model. phases were refined and extended from 4 a to 3 a resolution by means of the noncrystallographic molecular 222 symmetry. the r-factor was reduced to 28.7%, using 2.8 a resol ... | 1990 | 2330370 |
| metal ion dependence of phosphorothioate atp analogues in the bacillus stearothermophilus tyrosyl-trna synthetase reaction. | pre-steady-state kinetic analyses on the formation of tyrosyl adenylate from tyrosine and each of the four diastereomers of alpha- and beta-phosphorothioate adenosine triphosphates [atp alpha s and atp beta s; eckstein, f., & goody, r. (1976) biochemistry 15, 1685-1691; yee, d., armstrong, v. w., & eckstein, f. (1979) biochemistry 18, 4116-4123] were performed in the presence of mg2+, co2+, and cd2+ as the divalent metal ion cofactor. a modest preference of 5.5-fold in kappa 3/ka' (where kappa 3 ... | 1990 | 2334722 |
| composition of polar lipid acyl chains of bacillus stearothermophilus as affected by temperature and calcium. | bacillus stearothermophilus was grown within the temperature range of 48 to 68 degrees c in a complex medium and in the range of 45 to 72 degrees c in the presence of 2.5 mm ca2+. the main fatty acids of lipid extracts contain 15 to 17 carbon atoms, mostly branched-chain species. the most prevalent saturated straight-chain fatty acid is n-c16. the total amount of branched-chain species decreases with increasing temperature of growth from 48 to 68 degrees c, whereas the straight-chain species inc ... | 1990 | 2369582 |
| monitoring dental sterilizers' effectiveness using biological indicators. | the purpose of this study was to evaluate the effectiveness of dental office sterilizers as measured by their ability to kill bacterial spores present on biological indicator strips. the biological indicators used in this study contained two different spores, bacillus stearothermophilus and bacillus subtilis (spordi, amsco/medical products). ten spore test strips were sent to 87 dental offices; 51 sterilizers were tested. office personnel were instructed to place four strips in the center of a n ... | 1990 | 2370583 |
| [expression of xyle gene in bacillus stearothermophilus]. | using the gene expression vector pfdc11 for bacillus stearothermophilus cu21, a recombinant plasmid pfdx1 was constructed, which carries the pseudomonas-derived xyle gene coding for catechol 2,3-dioxygenase (cato2ase). cato2ase activity can be detected from cu21 (pfdx1) cells grown at 48 degrees c. this result indicates that the promoterless xyle gene can be expressed in a thermophilic host under the direction of a promoter from a thermophilic bacterium. by selecting kmr mutants at elevated grow ... | 1990 | 2372405 |
| studies on the interactions of glycerol dehydrogenase from bacillus stearothermophilus with zn2+ ions and nadh. | the interactions of the essential divalent cation, zn2+, with the binary complex formed between glycerol dehydrogenase (glycerol:nad+ 2-oxidoreductase, ec 1.1.1.6) and its coenzyme nadh have been examined by fluorescence spectroscopy. both the metallo and non-metallo form of the enzyme bind the coenzyme nadh. the addition of zn2+ ions to a solution of the binary complex formed between metal-depleted enzyme and nadh results in a rapid increase in fluorescence emission at 430 nm. this has been use ... | 1990 | 2378897 |
| nucleotide sequence of the alpha-amylase-pullulanase gene from clostridium thermohydrosulfuricum. | the nucleotide sequence of the gene (apu) encoding the thermostable alpha-amylase-pullulanase of clostridium thermohydrosulfuricum was determined. an open reading frame of 4425 bp was present. the deduced polypeptide (mr 165,600), including a 31 amino acid putative signal sequence, comprised 1475 amino acids, with no cysteine residues. the structural gene was preceded by the consensus promoter sequence ttgaca tataat, a putative regulatory sequence and a putative ribosome-binding sequence aaagggg ... | 1990 | 2391488 |
| [defective function of bioindicators for steam sterilization]. | it can be shown, that under certain conditions commercially available indicators with bacillus stearothermophilus and packages of native spores from soil prepared according to din 58 946/4 react differently to treatment in a lab-type steam sterilizer. the differences were most evident when incomplete evacuation of air had to be supposed. these results lead to the conclusion that some bioindicators are not able to show the inefficient function of steam sterilizers caused by local residuals of air ... | 1990 | 2393488 |
| bacillus thiaminolyticus sp. nov., nom. rev. | the name "bacillus thiaminolyticus" kuno 1951 was not included on the approved lists of bacterial names and has lost standing in bacteriological nomenclature. the genetic homogeneity of "bacillus thiaminolyticus" was assessed by determining guanine-plus-cytosine contents by the buoyant density method and by measuring dna relatedness by using spectrophotometric reassociation procedures. of the 26 strains which i studied, 24 had guanine-plus-cytosine contents in the range from 52 to 54 mol%. the c ... | 1990 | 2397192 |
| complete amino-acid sequence of glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic eubacterium thermotoga maritima. | 1. the complete amino-acid sequence of d-glyceraldehyde-3-phosphate dehydrogenase (gapdh) from the extreme thermophilic eubacterium thermotoga maritima has been determined by classical automated sequence analysis of peptides derived by chemical fragmentation with cyanogen bromide and enzymatic cleavages with specific proteases. 2. the protein contains 332 amino acids per subunit. its sequence is as follows: (sequence; see text) 3. comparing the given sequence with those of the enzymes from the m ... | 1990 | 2401296 |
| characterization of the bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in saccharomyces cerevisiae. | recombinant clones containing the manganese superoxide dismutase (mnsod) gene of bacillus stearothermophilus were isolated with an oligonucleotide probe designed to match a part of the previously determined amino acid sequence. complementation analyses, performed by introducing each plasmid into a superoxide dismutase-deficient mutant of escherichia coli, allowed us to define the region of dna which encodes the mnsod structural gene and to identify a promoter region immediately upstream from the ... | 1990 | 2407726 |
| complete nucleotide sequence of a gene coding for heat- and ph-stable alpha-amylase of bacillus licheniformis: comparison of the amino acid sequences of three bacterial liquefying alpha-amylases deduced from the dna sequences. | the gene coding for the heat-stable and ph-stable alpha-amylase of bacillus licheniformis 584 (atcc 27811) was cloned in escherichia coli and the nucleotide sequence of a dna fragment of 1,948 base pairs containing the entire amylase gene was determined. as inferred from the dna sequence, the b. licheniformis alpha-amylase had a signal peptide of 29 amino acid residues and the mature enzyme comprised 483 amino acid residues, giving a molecular weight of 55,200. the amino acid sequence of b. lich ... | 1985 | 2418011 |
| structural analysis of 5s rrna, 5s rrna-protein complexes and ribosomes employing rnase h and d(gttcgg). | the hybridization of d(gttcgg) to eubacterial 5s rrnas, 5s rrna-protein complexes, 70s ribosomes and 50s and 30s ribosomal subunits was investigated. this oligonucleotide, which may be considered to be an analogue of the t psi cg loop of trnas, was chosen in order to investigate a possible interaction between trnas with ribosomal components during protein synthesis. the hybridization was analysed by rnase h hydrolysis studies and, in the case of the ribosomes and ribosomal subunits, in addition ... | 1987 | 2434327 |
| the sequence of the 6s rna gene of pseudomonas aeruginosa. | from the gram-negative eubacterium pseudomonas aeruginosa we have isolated a stable 6s rna, approximately 180 nucleotides in length. the rna was partially sequenced and identified by comparison with the known escherichia coli 6s rna sequence. southern hybridizations revealed a single copy gene coding for the 6s rna. dna from other prokaryotes, i.e. e. coli, thermus thermophilus, bacillus subtilis, bacillus stearothermophilus and halobacterium maris mortui, did not give detectable hybridization s ... | 1987 | 2438656 |
| evidence for two activated forms of pyruvate kinase from bacillus stearothermophilus in the presence of ribose 5-phosphate. | allosteric activation of pyruvate kinase from a thermophilic bacterium, bacillus stearothermophilus, by ribose 5-phosphate (r5p) was kinetically examined. two activated forms of this enzyme could be distinguished, depending on the r5p concentration. one form (form i) was observed at about 10(-5) m r5p. it showed a slightly negative cooperativity for phosphoenolpyruvate (pep). the other form (form ii) was observed at more than 10(-3) m r5p and showed michaelis-menten kinetics for pep. the pep and ... | 1987 | 2439493 |
| molecular sieving through s layers of bacillus stearothermophilus strains. | the permeability properties and the exclusion limits of the crystalline surface layers (s layers) of two selected strains of bacillus stearothermophilus were investigated. measurements were performed of passive solute uptake into the intracellular space of native or glutaraldehyde-treated sacculi. native sacculi were prepared from whole cells by extracting the cytoplasmic membrane with triton x-100 under conditions which preserved the integrity of the s layer and the peptidoglycan-containing lay ... | 1987 | 2442137 |
| the secondary structure of ribonuclease p rna, the catalytic element of a ribonucleoprotein enzyme. | secondary structure models for the ribonuclease (rnaase) p rnas of bacillus subtilis and e. coli were derived by a phylogenetic comparative analysis of published sequences as well as four novel ones. the rnaase p rna genes from bacillus megaterium, bacillus brevis, bacillus stearothermophilus, and pseudomonas fluorescens were cloned, sequenced, and compared with the other available sequences. regions of pairing were identified by the occurrence of homologous complementary sequences that vary amo ... | 1988 | 2449969 |
| three-dimensional image reconstruction from ordered arrays of 70s ribosomes. | a better understanding of the molecular mechanism of protein biosynthesis still awaits a reliable model for the ribosomal particle. we describe here the application of a diffraction technique, namely three-dimensional image reconstruction from two-dimensional sheets of 70s ribosomes from bacillus stearothermophilus at 47 a resolution. the three-dimensional model obtained by these studies shows clearly the two subunits, the contact points between them, an empty space large enough to accommodate t ... | 1987 | 2450592 |
| in vitro incorporation of eubacterial, archaebacterial and eukaryotic 5s rrnas into large ribosomal subunits of bacillus stearothermophilus. | bacillus stearothermophilus large ribosomal subunits were reconstituted in the presence of 5s rrnas from different origins and tested for their biological activities. the results obtained have shown that eubacterial and archaebacterial 5s rrnas can easily substitute for b. stearothermophilus 5s rrna in the reconstitution, while eukaryotic 5s rrnas yield ribosomal subunits with reduced biological activities. from our results we propose an interaction between nucleotides 42-47 of 5s rrna and nucle ... | 1988 | 2453840 |
| three-dimensional location of ribosomal protein bl2 from bacillus stearothermophilus, a key component of the peptidyl transferase center. | protein bl2 from bacillus stearothermophilus has been localized by immunoelectron microscopy on the interface side of the 50 s subunit, beneath the angle formed between the central protuberance and the l1 protuberance. the immuno-electron microscopic data suggest that the interface region of the 50 s particle is not as flat as most of the proposed three-dimensional models suggest, but instead there is a significant concavity. since several studies demonstrated that bl2 is implicated in peptidyl ... | 1988 | 2454842 |
| effect of growth temperature and media composition on the fatty acid composition of bacillus stearothermophilus an 002. | the influence of growth temperature, media composition and cell age on the chemical composition of bacillus stearothermophilus strain an 002 has been determined. the total cellular protein decreased and the free amino acid content increased with growth temperature, in both exponential and stationary growth phase. the protein and free amino acid contents of cells were higher in the stationary phase than in the exponential phase, irrespective of growth temperature and media composition. the rna co ... | 1988 | 2455474 |
| crystals of wild-type, mutated, derivatized and complexed 50 s ribosomal subunits from bacillus stearothermophilus suitable for x-ray analysis. | three-dimensional single crystals of wild-type and mutated 50 s ribosomal subunits from bacillus stearothermophilus, as well as crystals of reconstituted subunits containing heavy-atom clusters and complexes of these subunits with trna and a short nascent polypeptide chain, were grown from polyethylene glycol in the presence of salts at low concentrations. within experimental error, all these crystals are isomorphous, packed with monoclinic symmetry (c2) in unit cells of a = 300 a, b = 546 a, c ... | 1989 | 2467005 |
| structural and kinetic bases for the recognition of trnatyr by tyrosyl-trna synthetase. | the aminoacylation of transfer rna is a key step of translation since it relates amino acids to anticodons. to understand how the tyrosyl-trna synthetase (tyrts) from bacillus stearothermophilus recognizes trna(tyr), we constructed 14 new mutant tyrts by site-directed mutagenesis, determined their kinetic properties and used these and previous data to construct a detailed structural model of the complex between tyrts and the acceptor arm of trna(tyr). in the model arg207, lys208, asn 146 and glu ... | 1989 | 2467006 |
| nucleotide sequence of the neopullulanase gene from bacillus stearothermophilus. | the gene (nplt) for a new type of pullulan-hydrolysing enzyme, neopullulanase, from bacillus stearothermophilus trs40 was sequenced. the dna sequence revealed only one large open reading frame, composed of 1764 bases and 588 amino acid residues (mr 69144). although the thermostable neopullulanase contained eight cysteine residues, they did not provide conformational stability by disulphide bonds. a comparison was made of the amino acid sequences of alpha-amylase, neopullulanase, isoamylase, pull ... | 1989 | 2482332 |
| expression of the insecticidal protein gene from bacillus thuringiensis subsp. aizawai in bacillus subtilis and in the thermophile bacillus stearothermophilus by using the alpha-amylase promoter of the thermophile. | expression of the insecticidal protein gene from bacillus thuringiensis subsp. aizawai ipl7 in b. subtilis mi113 and b. stearothermophilus sic1 was examined. production of the protein (130 kilodaltons [kda]) was analyzed by its reaction with antibody against the insecticidal proteins of the parental b. thuringiensis. when the original gene containing its own promoter was subcloned in b. subtilis, only a small amount of the protein was produced. therefore, both the promoter for the b. stearotherm ... | 1989 | 2482704 |
| expression and nucleotide sequence of the lactobacillus bulgaricus beta-galactosidase gene cloned in escherichia coli. | the lactobacillus bulgaricus beta-galactosidase gene was cloned on a ca. 7-kilobase-pair hindiii fragment in the vector pkk223-3 and expressed in escherichia coli by using its own promoter. the nucleotide sequence of the gene and approximately 400 bases of 3'- and 5'-flanking sequences was determined. the amino acid sequence of the beta-galactosidase, deduced from the nucleotide sequence of the gene, yielded a monomeric molecular mass of ca. 114 kilodaltons, slightly smaller than the e. coli lac ... | 1989 | 2492511 |
| isolation and characterisation of the glycerol dehydrogenase from bacillus stearothermophilus. | a protocol for the rapid purification of the glycerol dehydrogenase (glycerol: nad+ 2-oxidoreductase, ec 1.1.1.6) from the thermophile bacillus stearothermophilus has been developed using a combination of chromatographic techniques including affinity chromatography on a sepharose-immobilised triazine dye (procion red, he3b, ici). substrate specificity has been examined and km values determined. the protein has been shown to have an oligomeric mr of approx. 180,000 and consists of four identical ... | 1989 | 2493267 |
| mechanism of inactivation of alanine racemase by beta, beta, beta-trifluoroalanine. | the alanine racemases are a group of plp-dependent bacterial enzymes that catalyze the racemization of alanine, providing d-alanine for cell wall synthesis. inactivation of the alanine racemases from the gram-negative organism salmonella typhimurium and gram-positive organism bacillus stearothermophilus with beta, beta, beta-trifluoroalanine has been studied. the inactivation occurs with the same rate constant as that for formation of a broad 460-490-nm chromophore. loss of two fluoride ions per ... | 1989 | 2496744 |
| viable starter culture, beta-galactosidase activity, and lactose in duodenum after yogurt ingestion in lactase-deficient humans. | ten lactose malabsorbers were intubated and given fresh or heated yogurt to which polyethylene-glycol (peg) and spores of bacillus stearothermophilus (sbs) had been added as internal standards. in duodenal samples taken after fresh yogurt ingestion, viable starter culture was detected for 60 min in 6 of 7 subjects and the ratio of microbial beta-galactosidase activity to sbs remained similar during this period to its value in the preingested yogurt. in the two groups ingesting fresh and heated y ... | 1989 | 2497633 |
| on the effect on specificity of thr246----gly mutation in l-lactate dehydrogenase of bacillus sterothermophilus. | the function of the amino acid thr246 in l-lactate dehydrogenase from bacillus stearothermophilus has been investigated by site-directed replacement with glycine. kinetic experiments with a number of 2-oxo acids showed strongly reduced activity for the mutated enzyme. however, the mutant enzyme shows a relative preference for the large hydrophobic sidechains of alpha-keto acids and an even higher specific activity than the wild-type lactate dehydrogenase for the polar oxaloacetate substrate. gra ... | 1989 | 2499337 |
| activation of hageman factor and prekallikrein and generation of kinin by various microbial proteinases. | activation of the hageman factor-kallikrein-kinin system by serratial 56-kda proteinase was previously demonstrated (matsumoto, k., yamamoto, t., kamata, t., and maeda, h. (1984) j. biochem. (tokyo) 96, 739-749; kamata, r., yamamoto, t., matsumoto, k., and maeda, h. (1985) infect. immun. 48, 747-753). to investigate whether the activation of the system is specific for 56-kda proteinase or is found similarly with other microbial proteinases, 11 proteinases of microbial origins were studied; the 5 ... | 1989 | 2499581 |
| structural comparison of 26s rrna-binding ribosomal protein l25 from two different yeast strains and the equivalent proteins from three eubacteria and two chloroplasts. | the sequences of saccharomyces carlsbergensis ribosomal protein (r-protein) sl25* and its equivalents from candida utilis (cl25), escherichia coli (el23), bacillus stearothermophilus (bl23), mycoplasma capricolum (ml23), marchantia polymorpha chloroplasts (mcpl23), and nicotiana tabacum chloroplasts (ncpl23) were examined using a computer program that evaluates the extent of sequence similarity by calculating correlation coefficients for each pair of residues in two proteins from a number of phy ... | 1989 | 2501503 |
| site-directed mutagenesis of glyceraldehyde-3-phosphate dehydrogenase reveals the role of residue ser148. | a mutant of bacillus stearothermophilus d-glyceraldehyde-3-phosphate dehydrogenase, ser148----ala, was produced by oligonucleotide-directed mutagenesis. the study of the catalytic properties of this mutant has shown that this mutation significantly affects the michaelis constant of inorganic phosphate and to a lesser extent that of 1,3-diphosphoglycerate and d-glyceraldehyde-3-phosphate. this result is consistent with model-building studies which show that, for the phosphorylation step of cataly ... | 1989 | 2501780 |
| overproduction of thermostable leucine dehydrogenase of bacillus stearothermophilus and its one-step purification from recombinant cells of escherichia coli. | we have cloned the leucine dehydrogenase (ec 1.4.1.9) gene from a thermophile, bacillus stearothermophilus, into escherichia coli mv1184 with a vector plasmid, puc119. the cloned cells produced a large amount of the thermostable enzyme, which corresponds to about 60% of the total soluble protein. the enzyme was purified to more than 95% homogeneity by only one step, heat treatment of the cell-extracts, with an average yield of 75 mg/g of wet cells (obtained from 100 ml of the culture). | 1989 | 2503013 |
| structure of tyrosyl-trna synthetase refined at 2.3 a resolution. interaction of the enzyme with the tyrosyl adenylate intermediate. | the crystal structure of tyrosyl-trna synthetase (ec 6.1.1.1) from bacillus stearothermophilus has been refined to a crystallographic r-factor of 22.6% at 2.3 a resolution using a restrained least-squares procedure. in the final model the root-mean-square deviation from ideality for bond distances is 0.018 a and for angle distances is 0.044 a. each monomer consists of three domains: an alpha/beta domain (residues 1 to 220) containing a six-stranded beta-sheet, an alpha-helical domain (248 to 318 ... | 1989 | 2504923 |
| site-directed mutagenesis in bacillus stearothermophilus fructose-6-phosphate 1-kinase. mutation at the substrate-binding site affects allosteric behavior. | arg252 of fructose-6-phosphate 1-kinase (pfk) from bacillus stearothermophilus has been proposed to be involved in the binding of the substrate fru-6-p. we demonstrate here that mutation of this residue to alanine converts the enzyme to a form with characteristics similar to those of its allosterically tight form. the mutant enzyme exhibits a high affinity for its inhibitor phosphoenolpyruvate (a 68-fold difference compared to wild type) and a dramatically decreased fru-6-p affinity (1500-fold i ... | 1989 | 2521215 |
| serine and tyrosine protein kinase activities in streptococcus pyogenes. phosphorylation of native and synthetic peptides of streptococcal m proteins. | two forms of protein kinase activity were isolated from crude extracts of streptococcus pyogenes and partially purified by ion exchange chromatography and affinity chromatography. the phosphorylation activities were shown to be insensitive to camp, required the presence of divalent cations, and eluted from a sephadex g-200 column with approximate molecular masses of 60 and 45 kda, respectively. both enzymes were capable of phosphorylating eukaryotic proteins and synthetic polypeptides in additio ... | 1989 | 2521633 |
| crystal structure of unliganded phosphofructokinase from escherichia coli. | in an attempt to characterize the mechanism of co-operativity in the allosteric enzyme phosphofructokinase from escherichia coli, crystals were grown in the absence of activating ligands. the crystal structure was determined to a resolution of 2.4 a by the method of molecular replacement, using the known structure of the liganded active state as a starting model, and has been refined to a crystallographic r-factor of 0.168 for all data. although the crystallization solution would be expected to ... | 1989 | 2527305 |
| complete nucleotide sequences of two phosphoglucoisomerase isozymes from bacillus stearothermophilus. | | 1989 | 2532320 |
| characterization and application of a thermostable primary transport system: cytochrome-c oxidase from bacillus stearothermophilus. | cytochrome-c oxidase from bacillus stearothermophilus has been purified to homogeneity by detergent extraction followed by deae-cellulose, hydroxyapatite- and gel-filtration chromatography. the enzyme is a typical cytochrome-aa3-type oxidase which binds carbon monoxide and is sensitive to classical oxidase inhibitors like cyanide and azide. the purified enzyme is composed of three different subunits (57, 37 and 22 kda). the subunit with intermediate molecular mass contains a covalently attached ... | 1989 | 2536327 |
| structural analysis of prokaryotic and eukaryotic 5s rrnas by rnase h. | the availabilities of single-stranded 5s rrna regions c, d and d' for base pairing interactions were analyzed by using synthetic dna oligomers. hybrid formation was detected by the endonucleolytical mode of the rna-dna specific action of rnase h. provided that the hybrid interaction involved 6 successive base pairs, 5s rrna loop c nucleotides 42-47 displayed accessibility in escherichia coli, bacillus stearothermophilus and thermus thermophilus 5s rrnas as well as in eukaryotic 5s rrnas from sac ... | 1989 | 2561346 |
| mechanism of l-glutamate transport in membrane vesicles from bacillus stearothermophilus. | in the presence of electrochemical energy, several branched-chain neutral and acidic amino acids were found to accumulate in membrane vesicles of bacillus stearothermophilus. the membrane vesicles contained a stereo-specific transport system for the acidic amino acids l-glutamate and l-aspartate, which could not translocate their respective amines, l-glutamine and l-asparagine. the transport system was thermostable (ti = 70 degrees c) and showed highest activities at elevated temperatures (60 to ... | 1989 | 2563364 |
| characterization of amino acid transport in membrane vesicles from the thermophilic fermentative bacterium clostridium fervidus. | amino acid transport was studied in membrane vesicles of the thermophilic anaerobic bacterium clostridium fervidus. neutral, acidic, and basic as well as aromatic amino acids were transported at 40 degrees c upon the imposition of an artificial membrane potential (delta psi) and a chemical gradient of sodium ions (delta microna+). the presence of sodium ions was essential for the uptake of amino acids, and imposition of a chemical gradient of sodium ions alone was sufficient to drive amino acid ... | 1989 | 2567728 |
| the effect of air on the moist-heat resistance of bacillus stearothermophilus spores. | the presence of air in an autoclave chamber and load is generally considered to reduce the lethal effect of the process on bacterial spores. in this study, the heat inactivation of spores of bacillus stearothermophilus (ncib 8224) was measured in the presence and absence of air at 100% relative humidity. the results confirm that air significantly enhances the lethality of moist-heat on this strain of b. stearothermophilus. | 1989 | 2575633 |
| dynamic sterilization of titanium implants with ultraviolet light. | all implantable devices must be sterile. however, autoclaves produce poor surface properties that jeopardize the integration process. the application of a modified ultraviolet light source has proven to enhance bioreactivity by controlling surface properties, but it lacks validation of its sterilization capabilities. forty-eight titanium implants were contaminated with spores of the biological indicator bacillus stearothermophilus and subjected to "dynamic sterilization" by ultraviolet light. fo ... | 1989 | 2599585 |
| relationship between the digestive microflora of the newborn and maternal microflora in rodents as evidenced by a transit marker. | the cutaneous and digestive microflora of the dam, constitute the main sources of bacteria likely to colonize the digestive tract of the newborn. our objective was to study the conditions (delay, level) in relation to the maternal microflora. spores of bacillus stearothermophilus were used as a tracer. they were spread on the mother's udder or given per os to the mother. in our study, we used holoxenic rats and axenic mice. the tracer was then followed in newborn rats and newborn mice. transfer ... | 1989 | 2619205 |
| protoplast transformation of bacillus stearothermophilus nub36 by plasmid dna. | an efficient protoplast transformation system was established for bacillus stearothermophilus nub3621 using thermophilic plasmid ptht15 tcr (4.5 kb) and mesophilic plasmid plw05 cmr (3 kb), a spontaneous deletion derivative of ppl401 cmr kmr. the efficiency of transformation of nub3621 with plw05 and ptht15 was 2 x 10(7) to 4 x 10(8) transformants per micrograms dna. the transformation frequency (transformants per regenerant) was 0.5 to 1.0. chloramphenicol-resistant and tetracycline-resistant t ... | 1989 | 2621450 |
| [isolation, purification and properties of restrictase and methylase bstn1 from bacillus stearothermophilus]. | restriction-methylation enzymes bstn1 from bacillus stearothermophilus were isolated and purified. these enzymes are related to a new class of restriction-methylation enzymes of the second type, whose modifying component is n4-cytosine-dna-methylase. both enzymes recognize the dna sequence cc(a/t)gg. restrictase bstn1 is a protein made up of one subunit with a molecular mass of 25 kda. the molecular mass of native dna-methylase bstn1 is about 55 kda. the temperature optima for restrictase and me ... | 1989 | 2627557 |
| lactoferrin and lysozyme in milk during acute mastitis and their inhibitory effect in delvotest p. | microbiological methods for detection of antibiotic residues in milk give no explanations regarding the identity of the inhibitory substance(s). natural antibacterial substances, present at higher concentrations in mastitic milk and in colostrum, occasionally cause false positive results in antibiotic assays. in an earlier investigation, lysozyme and lactoferrin were shown to inhibit the growth of bacillus stearothermophilus var. calidolactis spores, used as test organism in delvotest p. to stud ... | 1989 | 2628440 |
| structure and properties of malic enzyme from bacillus stearothermophilus. | the malic enzyme (ec 1.1.1.38) gene of bacillus stearothermophilus was cloned in escherichia coli, and the enzyme was purified to homogeneity from the e. coli clone. in addition to the nad(p)-dependent oxidative decarboxylation of l-malate, the enzyme catalyzes the decarboxylation of oxalacetate. the enzyme is a tetramer of mr 200,000 consisting of four identical subunits of mr 50,000. the ph optima for malate oxidation and pyruvate reduction are 8.0 and 6.0, respectively; and the optimum temper ... | 1989 | 2644282 |
| the complete amino acid sequence of ribosomal protein s18 from the moderate thermophile bacillus stearothermophilus. | the amino acid sequence of ribosomal protein s18 from bacillus stearothermophilus has been completely determined by automated sequence analysis of the intact protein as well as of peptides derived from digestion with staphylococcus aureus protease at ph 4.0 and cleavage with cyanogen bromide. the carboxy-terminal region was verified by both amino acid analyses of chymotryptic peptides and by mass spectrometry from the terminal region. the protein contains 77 amino acid residues and has an mr of ... | 1989 | 2647521 |
| nucleotide sequence determination of the dna region coding for bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase and of the flanking dna regions required for its expression in escherichia coli. | the complete nucleotide sequence of a 3541-base pairs (bp) dna fragment from bacillus stearothermophilus able to complement an escherichia coli glyceraldehyde-3-phosphate-dehydrogenase (gapdh) mutant (gapd-) has been determined. the b. stearothermophilus gap gene consists of a 1005-bp open reading frame commencing with an atg start codon and ending with a taa stop codon. upstream from the start codon is a strong shine-dalgarno sequence typical of gram-positive bacteria. only one putative rna pol ... | 1989 | 2656407 |
| probing the substrate-binding sites of aminoacyl-trna synthetases with the procion dye green he-4bd. | a reactive bis-dichloro derivative of the procion dye green he-4bd was shown to inactivate irreversibly methionyl-trna synthetase (mts) from escherichia coli and also tryptophyl-trna synthetase (wts) and tyrosyl-trna synthetase (yts) from bacillus stearothermophilus at ph 8.5 and 37 degrees c. at a 5-fold excess of reactive dye over enzyme subunit concentration mts was quantitatively inactivated within 20 min in the atp/pyrophosphate exchange assay, whereas wts and yts show an 80% loss of activi ... | 1989 | 2658972 |
| (1-aminoethyl)boronic acid: a novel inhibitor for bacillus stearothermophilus alanine racemase and salmonella typhimurium d-alanine:d-alanine ligase (adp-forming). | (1-aminoethyl)boronic acid (ala-b), an analogue of alanine in which a boronic acid group replaces the carboxyl group, has been synthesized and found to inhibit the first two enzymes, alanine racemase (from bacillus stearothermophilus, ec 5.1.1.1) and d-alanine:d-alanine ligase (adp-forming) (from salmonella typhimurium, ec 6.3.2.4), of the d-alanine branch of bacterial peptidoglycan biosynthesis. in both cases, time-dependent, slow binding inhibition is observed due to the generation of long-liv ... | 1989 | 2663072 |
| interactions of escherichia coli so-187 trna(ival) with bacillus stearothermophilus valine-trna synthetase studied by 13c-nmr. | uracil isotopically labelled with 13c at c4 and c5 has been incorporated into nucleic acids of the escherichia coli uracil auxotroph, so-187. [4,5-13c]uracil-labeled trna(ival) was isolated and purified. 13c longitudinal relaxation times measured at 67.8 mhz demonstrated that the c5 dipole caused a 20-50% increase in the c4 relaxation. interactions of this trna with valine-trna synthetase (vts) purified from bacillus stearothermophilus were established by 13c-nmr. specific spectral changes were ... | 1989 | 2667642 |
| replacement of potassium ions by ammonium ions in different micro-organisms grown in potassium-limited chemostat culture. | the biomass concentration extant in potassium-limited cultures of either klebsiella pneumoniae or bacillus stearothermophilus (when growing at a fixed temperature and dilution rate in a glucose/ammonium salts medium) increased progressively as the medium ph value was raised step-wise from 7.0 to 8.5. because the macromolecular composition of the organisms did not vary significantly, this increase in biomass could not be attributed to an accumulation of storage-type polymers but appeared to refle ... | 1989 | 2669673 |
| cloning, nucleotide sequence, and expression in escherichia coli of the bacillus stearothermophilus peroxidase gene (pera). | the gene encoding a thermostable peroxidase was cloned from the chromosomal dna of bacillus stearothermophilus iam11001 in escherichia coli. the nucleotide sequence of the 3.1-kilobase ecori fragment containing the peroxidase gene (pera) and its flanking region was determined. a 2,193-base-pair open reading frame encoding a peroxidase of 731 amino acid residues (mr, 82,963) was observed. a shine-dalgarno sequence was found 9 base pairs upstream from the translational starting site. the deduced a ... | 1989 | 2670897 |
| addition of a methyl group changes both the catalytic velocity and thermostability of the neutral protease from bacillus stearothermophilus. | specific activity was compared between wild-type (wt) neutral protease from bacillus stearothermophilus and mutant protease (m1; gly144 replaced by ala144) with enhanced thermostability. when casein was used as a substrate, m1 showed 1.5-times higher specific activity than that of wt. in contrast, the specific activities of m1 for soluble reduced lysozyme and insulin b chain were lower than those of wt by 17.2 and 13.2%, respectively. after digestion of the insulin a chain by these enzymes, the ... | 1989 | 2673840 |
| integration and expression of alpha-amylase and endoglucanase genes in the lactobacillus plantarum chromosome. | a commercial grass silage starter strain of lactobacillus plantarum was transformed by high-frequency electroporation with plasmids containing an alpha-amylase gene from bacillus stearothermophilus and an endoglucanase gene from clostridium thermocellum. both genes were expressed from their native regulatory signals, and active enzymes were found in the supernatant. however, the segregational stability of the transforming plasmids was rather low. therefore, the transforming genes were inserted i ... | 1989 | 2679379 |
| nucleotide sequences of genes encoding heat-stable and heat-labile glyceraldehyde-3-phosphate dehydrogenases; amino acid sequence and protein thermostability. | the structural gene (gapst) encoding glyceraldehyde-3-phosphate dehydrogenase (gpdh; ec 1.2.1.12) from bacillus stearothermophilus has been cloned in escherichia coli using plasmid pbr322 as a vector; the homologous gene (gapco) from bacillus coagulans was cloned from a phage lambda library. expression of the cloned gap genes revealed that, as in the wild-type (wt) organisms, the gpdh from b. stearothermophilus (gpdh-st) was intrinsically heat stable (hs) and that from b. coagulans (gpdh-co) hea ... | 1989 | 2684782 |
| protection of an unstable reaction intermediate examined with linear free energy relationships in tyrosyl-trna synthetase. | linear free energy relationships (lfers) are powerful tools in the search to understand the relationship between molecular structure and activity. they frequently link the changes in the rate constants for a reaction to changes in the equilibrium constant caused by alterations in structure. in physical-organic chemistry, these have been interpreted to give information on the structure of the transition state. similar phenomena have been observed for reactions catalyzed by a series of engineered ... | 1989 | 2690955 |
| sequence and characteristics of the bifidobacterium longum gene encoding l-lactate dehydrogenase and the primary structure of the enzyme: a new feature of the allosteric site. | the gene ldh, encoding l-lactate dehydrogenase (ldh; ec 1.1.1.27) of bifidobacterium longum am101-2, was cloned in escherichia coli using an oligodeoxyribonucleotide hybridization probe. the amino acid (aa) sequence, deduced from the sequence of the cloned dna, was consistent with the results of protein chemical analysis of b. longum ldh. the transcription start points (tsp) in b. longum were identified by s1 nuclease mapping. a sequence, gtagcaa-(14 bp)-ttataga, which is located a few bp upstre ... | 1989 | 2695396 |
| 4-chloroacetylpyridine adenine dinucleotide. a highly reactive and chromophoric affinity label of glyceraldehyde-3-phosphate dehydrogenase from sturgeon. | the analogue of nad+, 4-chloroacetylpyridine-adenine dinucleotide (clac4pdad+), inactivated the glyceraldehyde-3-phosphate dehydrogenase from sturgeon at a high rate. an affinity labeling was shown to occur with clac4pdad+. the mononucleotide 4-chloroacetylpyridine 1-beta-d-ribose 5'-phosphate (clac4pdmn+) reacted with the enzyme in a second-order reaction whose rate was much smaller than that calculated for clac4pdad+ taken as a second-order rate reagent. the rate of the reaction of clac4pdad+ ... | 1989 | 2714279 |
| molecular and immunological characterization of plastid and cytosolic pyruvate kinase isozymes from castor-oil-plant endosperm and leaf. | 1. monospecific antiserum was raised in rabbits to homogeneous cytosolic pyruvate kinase isolated from 5-day-old germinating endosperm of the castor oil plant, ricinus communis. an earlier study demonstrated that the purified enzyme is putatively heterotetrameric, composed of two subunits which migrate as 57-kda and 56-kda proteins upon sodium dodecyl sulfate/polyacrylamide gel electrophoresis [plaxton, w. c. (1988) plant physiol. (bethesda) 86, 1065-1069]. both proteins were detected on western ... | 1989 | 2714295 |
| isolation of phenol-degrading bacillus stearothermophilus and partial characterization of the phenol hydroxylase. | bacillus stearothermophilus br219, isolated from river sediment, degraded phenol at levels to 15 mm at a rate of 0.85 mumol/h (4 x 10(6) cells). the solubilized phenol hydroxylase was nadh dependent, exhibited a 55 degrees c temperature optimum for activity, and was not inhibited by 0.5 mm phenol. | 1989 | 2719481 |
| [sterilization of non-air evacuated infusion flasks containing little water]. | studies have been carried out in connection with the sterilization of glass infusion bottles from which air had not been evacuated and containing certain limited quantities of water. bottles with a capacity of 500 and 1000 cm3 were filled with 1-3 cm3 distilled water and sterilized in an autoclave for 20 min. set at a temperature of 121 degrees c reaching up to 126 degrees c. the sterility was tested by introducing bags containing bacillus stearothermophilus spores into the centre of the bottles ... | 1989 | 2726985 |
| catalytic properties of thermophilic lactate dehydrogenase and halophilic malate dehydrogenase at high temperature and low water activity. | thermophilic lactate dehydrogenases from thermotoga maritima and bacillus stearothermophilus are stable up to temperature limits close to the optimum growth temperature of their parent organisms. their catalytic properties are anomalous in that km shows a drastic increase with increasing temperature. at low temperatures, the effect levels off. extreme halophilic malate dehydrogenase from halobacterium marismortui exhibits a similar anomaly. increasing salt concentration (nacl) leads to an optimu ... | 1989 | 2753046 |
| cloning and characterization of a gene cluster from bacillus stearothermophilus comprising infc, rpmi and rplt. | using two synthetic deoxyribonucleotide probes encoding segments of the primary structure of initiation factor if3 from bacillus stearothermophilus, we identified and cloned a segment of dna which carries the infc gene. as in escherichia coli, the infc gene begins with the unusual initiation triplet auu, and is followed by the structural genes for ribosomal proteins l35 and l20 (rpmi and rplt, respectively). | 1989 | 2779520 |
| multiple amylase genes in two strains of bacillus stearothermophilus. | plasmid dna fragments from bacillus stearothermophilus atcc29609 (br135) and chromosomal dna fragments from b. stearothermophilus atcc31195 (br132) were cloned into pbr322 and transformed into escherichia coli strain hb101. clones were selected which demonstrate extracellular expression of thermostable amylase. the dna inserts from both strains showed similar restriction maps. examination of the parental strains revealed plasmids of approx. 100 kb and 35 kb for br135 and no discernible plasmids ... | 1989 | 2787266 |
| on the effects of replacing the carboxylate-binding arginine-171 by hydrophobic tyrosine or tryptophan residues in the l-lactate dehydrogenase from bacillus stearothermophilus. | for l-lactate dehydrogenases (ldh's), the interaction of the guanidinium group of their arg 171 residue with the carboxylate group of an alpha-keto acid is of primary importance in orienting the substrate productively at the active site. ldh's such as that of bacillus stearothermophilus (bsldh) are of practical importance for the preparation of chiral 2-hydroxy acids used as synthons in asymmetric synthesis but would even be more valuable in this regard if their specificities were broader. with ... | 1989 | 2790015 |
| phosphorescence properties of trp-84 and trp-310 in glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | the phosphorescence spectra of trp-84 and trp-310 in glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus in an aqueous glass show distinct 0,0 vibrational bands with peaks at 406.5 and 410.5 nm. with the aid of external heavy-atom perturbation of iodide and the thermal quenching profile, it is concluded that although both chromophores are effectively buried, only one, viz., the 406.5 nm component, is embedded in a sufficiently rigid core of the protein to phosphoresce in fl ... | 1989 | 2804244 |
| subchronic toxicity studies in dogs and in utero rats fed diets containing bacillus stearothermophilus alpha-amylase from a natural or recombinant dna host. | beagle dogs and fischer 344 rats were fed diets containing 0, 36 or 72 units bacillus stearothermophilus alpha-amylase (bsa)/g food or of bacillus subtilis alpha-amylase (cbsa)/g food. the dogs (four/sex/group) received treated diets for 13 wk. for the rat studies, the parental (f0) generation (12 males and 24 females/group for the bsa study, and 26 rats/sex/group for the cbsa study) received treated diets for 13 or 4 wk, respectively, before breeding and through weaning of the f1 pups; 20 f1 ra ... | 1989 | 2807104 |
| novel procedures for derivatization of ribosomes for crystallographic studies. | undecagold and tetrairidium clusters have been used for the preparation of heavy-metal derivatives of ribosomal particles, necessary for the evaluation of phases in the x-ray structure determination of these large particles. to obtain specific binding, monofunctional reagents of the clusters were prepared and were covalently bound to free sulfhydryl groups on the surface of the ribosome. in addition, a mutant of bacillus stearothermophilus which lacks one ribosomal protein (bl11) was grown. the ... | 1989 | 2808418 |
| effects of engineering complementary charged residues into the hydrophobic subunit interface of tyrosyl-trna synthetase. appendix: kinetic analysis of dimeric enzymes that reversibly dissociate into inactive subunits. | wild-type tyrosyl-trna synthetase (tyrts) from bacillus stearothermophilus is a symmetrical dimer. four different heterodimeric enzymes have been produced by site-directed mutagenesis at the subunit interface so that the monomers are linked by a potential salt bridge in a hydrophobic environment. the two phe-164 residues of wild-type tyrts are on the axis of symmetry and interact in a hydrophobic region of the subunit interface. mutation of phe-164 to aspartate or glutamate in full-length tyrts ... | 1987 | 2820484 |
| the inhibition of glucokinase and glycerokinase from bacillus stearothermophilus by the triazine dye procion blue mx-3g. | glucokinase from bacillus stearothermophilus was irreversibly inactivated by the reactive dichlorotriazinyl dye procion blue mx-3g at ph 8.0. the enzyme was protected from inactivation by the substrate mgatp. kinetic data implied that the dye occupied the mgatp-binding site. the apparent km values for mgatp and d-glucose were found to be 70 microm and 210 microm respectively, and the kd of the pure reactive dye was 16 microm; 1 mol of the pure reactive dye bound to 1 mol of glucokinase subunit. ... | 1987 | 2823796 |
| nucleotide sequence and linkage map position of the genes for ribosomal proteins l14 and s8 in the maize chloroplast genome. | the nucleotide sequence of a 1287-base-pair segment of the maize (zea mays) chloroplast dna, encoding chloroplast ribosomal proteins l14, s8 and the c-terminal part of l16, has been determined using the dideoxy-chain-termination method. these data from a monocot plant are compared to the corresponding data from a dicot and a lower plant and from two bacteria. the deduced amino acid sequence of maize chloroplast l14 shows 80%, 81%, 51% and 52% and that of s8 shows 75%, 58%, 39% and 38% sequence i ... | 1988 | 2828044 |
| thermostability and superhelicity of plasmid dna in bacillus stearothermophilus. | the thermostability of the staphylococcal plasmids pc194 and pub110 and their antibiotic-resistance determinants was examined upon transfer to bacillus stearothermophilus cu21. plasmid pgs13, a pub110 derivative carrying the chloramphenicol acetyltransferase (cat) gene of pc194, could be maintained up to the maximum growth temperature (68 degrees c) by selection for chloramphenicol resistance. in the absence of selective pressure, pgs13 was lost at temperatures above 60 degrees c. segregational ... | 1987 | 2828885 |
| nucleotide sequence and linkage map position of the secx gene in maize chloroplast and evidence that it encodes a protein belonging to the 50s ribosomal subunit. | the nucleotide sequence of the segment of maize chloroplast dna lying between the map coordinate positions 32.59 and 32.98 kb and containing the secx gene has been determined. the derived amino acid sequence of maize chloroplast secx is 95%, 87% and 62% identical to the corresponding derived amino acid sequences from two plant chloroplasts and escherichia coli, respectively. it is also 70% identical to the experimentally determined amino acid sequence of a protein isolated from bacillus stearoth ... | 1987 | 2829909 |
| thermostable alanine racemase from bacillus stearothermophilus: dna and protein sequence determination and secondary structure prediction. | the nucleotide sequence of the alanine racemase (ec 5.1.1.1) gene from a thermophile, bacillus stearothermophilus, was determined by the dideoxy chain termination method with universal and synthetic site-specific primers. the amino acid sequence of the enzyme predicted from the nucleotide sequence was confirmed by peptide sequence information derived from the n-terminal amino acid residues and several tryptic fragments. the alanine racemase gene consists of 1158 base pairs encoding a protein of ... | 1988 | 2835089 |
| cation-induced thermostability of yeast and escherichia coli pyrophosphatases. | inorganic pyrophosphatases (ppiases) from both yeast and escherichia coli were found to be stable against heat denaturation in the presence of mg2+, as previously observed with the enzymes from thermophilic bacteria. no loss of activity was observed after 1 h of incubation at 50 degrees c and phs between 6 and 9 in the yeast enzyme, and at 60 degrees c and phs between 7.2 and 9.2 in the e. coli enzyme. such an induced thermostability of the e. coli enzyme was detected when mn2+, co2+, ca2+, cd2+ ... | 1988 | 2835971 |
| bsri, a unique restriction endonuclease from bacillus stearothermophilus which recognizes 5' actgg3'. | | 1988 | 2838810 |
| isolation and characterization of restriction endonuclease bstyi from bacillus stearothermophilus y406. | bstyi, an isoschizomer of xhoii and mfli, has been purified from bacillus stearothermophilus y406. this enzyme recognized 5'...pu/gatcpy...3' in dna and cleaved between pu and g in this sequence. bstyi can be easily isolated and purified by heparin-agarose column chromatography in a high yield (8000 units bstyi can be obtained per g wet wt of cells). | 1988 | 2839359 |
| transduction in bacillus stearothermophilus. | temperate and virulent bacteriophages isolated from soil were shown to carry out generalized transduction of bacillus stearothermophilus nub36. a transducing frequency of 1 x 10(-5) to 7 x 10(-4) was obtained for temperate phages tp-42 and tp-56. the transducing frequency for virulent phage tp-68 was two to three orders of magnitude lower. cotransfer analysis with the three phages showed that hom-1 is linked to thr-1 and that gly-1 is linked to his-1. | 1988 | 2841302 |
| [a new type of cleavage of the recognition site by the site-specific endonuclease bst 4.4i from bacillus stearothermophilus 4.4]. | a site-specific endonuclease bst 4.4i was isolated from the cell extract of bacillus stearothermophilus 4.4 and partially purified by chromatography on ultragel aca-44 and heparin-sepharose. it was shown that the endonuclease cleaves lambda and m13 dna yielding distinct fragments just as endonucleases of ii and iii types but, in contrast to them can produce two two-strand cuts separated with 30 to 32 nucleotides in the region of the recognition site. | 1988 | 2847759 |
| hydrogen-1 nuclear magnetic resonance of the nitrogenase iron protein (cp2) from clostridium pasteurianum. | proton nmr spectra (250 mhz) of the nitrogenase iron protein from clostridium pasteurianum (cp2) were found to display 9 or 10 paramagnetically shifted resonances in the 15-50 ppm range. the most shifted resonances belonged to two approximately equal subsets having temperature dependences of opposite sign. the latter occurrence is consistent with the interaction of the corresponding protons with an antiferromagnetically coupled metal center. the number of proton resonances of cp2, their position ... | 1988 | 2847787 |
| cloning and characterization of the gene for a methanol-utilising alcohol dehydrogenase from bacillus stearothermophilus. | the cloning and characterization of the alcohol dehydrogenase (adh) gene (adh) from bacillus stearothermophilus strain dsm2334, an obligate aerobe, are described. the clone directed the synthesis of adh as judged on western blots, activity gels and tetrazolium plates. it specified an enzyme that oxidised methanol as well as ethanol. the enzyme was found to be encoded by a single gene in b. stearothermophilus which did not cross-hybridize to adh clones from escherichia coli, yeast or maize. the c ... | 1988 | 2851486 |
| the activity of glutaraldehyde against clostridium difficile. | the sporicidal activity of 2% glutaraldehyde against bacillus subtilis var. globigii and bacillus stearothermophilus was compared with that against clostridium difficile. the aerobic species, normally chosen for test purposes, survived for 2 h but cl. difficile was killed in under 10 min. in view of the impractical and lengthy immersions required to satisfy standard tests using non-pathogenic spores, a less stringent standard would probably be more appropriate. | 1985 | 2859321 |
| excretion of penicillins and cephalexin in bovine milk following intramammary administration. | the elimination into bovine milk of beta-lactam antibiotic residues (procaine penicillin g, cloxacillin, ampicillin, oxacillin, cephalexin) following intramammary administration of 10 preparations marketed in france was studied. the quantitative analysis of residues was carried out by a microbiological agar diffusion method using bacillus stearothermophilus. sensitivity ranged from 0.001 i.u./ml for procaine penicillin g and 0.001 micrograms/ml for ampicillin to 0.02 micrograms/ml for cephalexin ... | 1989 | 2912796 |
| pattern of action of bacillus stearothermophilus neopullulanase on pullulan. | the action of neopullulanase from bacillus stearothermophilus on many oligosaccharides was tested. the enzyme hydrolyzed not only alpha-(1----4)-glucosidic linkages but also specific alpha-(1----6)-glucosidic linkages of several branched oligosaccharides. when pullulan was used as a substrate, panose, maltose, and glucose, in that order, were produced as final products at a final molar ratio of 3:1:1. according to these results, we proposed a model for the pattern of action of neopullulanase on ... | 1989 | 2914851 |
| nucleotide sequence of the phosphofructokinase gene from bacillus stearothermophilus and comparison with the homologous escherichia coli gene. | the gene (bs-pfk) for phosphofructokinase (pfk) from bacillus stearothermophilus has been cloned and sequenced. the deduced amino acid sequence is nearly identical to the sequence which was previously determined by peptide analysis. the elevated g + c content of bs-pfk relative to the homologous ec-pfka from escherichia coli is consistent with previous observations concerning genes from thermophilic prokaryotes. a significant degree of homology exists when the deduced amino acid sequence of b. s ... | 1987 | 2956156 |
| high-level expression of bacillus stearothermophilus 6-phosphofructo-1-kinase in escherichia coli. | the 6-phosphofructo-1-kinase (pfk) gene from bacillus stearothermophilus has been expressed at high levels in escherichia coli. this expression has been demonstrated by complementation studies, sodium dodecyl sulfate (sds)-polyacrylamide gel electrophoresis, and pfk assays of cell extracts. a level of b. stearothermophilus pfk expression corresponding to 20% of the total extracted protein was calculated from densitometric scans of an sds-polyacrylamide gel. the high level of recombinant gene exp ... | 1987 | 2963782 |
| site-directed mutagenesis at the regulatory site of fructose 6-phosphate-1-kinase from bacillus stearothermophilus. | we have mutated arg-25, asp-59 and arg-211 to alanine; and asp-59 also to methionine, in fructose 6-phosphate-1-kinase from b. stearothermophilus (designated as ra25, da59, ra211 and dm59 respectively). all four mutants did not change the affinity of the enzyme for atp. ra25 has half the affinity for fructose 6-phosphate and exhibits sigmoidicity with respect to this substrate (hill # = 2.0). da59 has the same affinity for phosphoenolpyruvate (pep) as the wild type whereas dm59 has 3-fold the af ... | 1988 | 2972287 |
| crystallization of and preliminary crystallographic data for bacillus stearothermophilus cyclodextrin glucanotransferase. | cyclodextrin glucanotransferase from bacillus stearothermophilus tc-91 has been crystallized from an ammonium sulfate solution by the dialysis equilibrium method. the crystals belong to the orthorhombic system, space group p2(1)2(1)2(1), with cell dimensions of a = 125.5 a, b = 88.1 a, and c = 81.5 a. the crystals appear to be suitable for x-ray structure analysis, diffracting to at least 2.1 a and being resistant to radiation damage. | 1988 | 2975653 |
| crystal structure of the complex of phosphofructokinase from escherichia coli with its reaction products. | the crystal structure of escherichia coli phosphofructokinase complexed with its reaction products fructose 1,6-bisphosphate (fru1,6p) and adp/mg2+, and the allosteric activator adp/mg2+, has been determined at 2.4 a resolution. the structure was solved by molecular replacement using the known structure of bacillus stearothermophilus phosphofructokinase, and has been refined to a crystallographic r-factor of 0.165 for all data. the crystallization mixture contained the substrate fructose 6-phosp ... | 1988 | 2975709 |