| synthesis of p1,p4-di(adenosine 5'-) tetraphosphate by leucyl-trna synthetase, coupled with atp regeneration. | a simple and practical procedure for the synthesis of p1,p4-di(adenosine 5'-) tetraphosphate from atp by the catalysis of leucyl-trna synthetase from bacillus stearothermophilus is described. km for leucine was 6.7 microm and for atp was 3.3 mm. the reaction yielded not only diadenosine tetraphosphate, but various byproducts such as p1,p3-(diadenosine 5'-) triphosphate, adp and amp. by coupling the reaction with an atp regeneration system by acetate kinase and adenylate kinase with acetylphospha ... | 1987 | 3606615 |
| the importance of arginine 171 in substrate binding by bacillus stearothermophilus lactate dehydrogenase. | a variant of lactate dehydrogenase from bacillus stearothermophilus has been engineered by site-directed mutagenesis in which an active-site arginine residue at position 171 in the protein sequence is replaced by lysine. replacement of this arginine by lysine has no effect on co-enzyme binding, a relatively small effect on the rate of turnover of the enzyme, but causes a 2000-fold increase in the michaelis constant for pyruvate, a 6000-fold increase in the dissociation constant for oxamate and r ... | 1987 | 3606622 |
| the complete amino acid sequence of the ribosomal 'a' protein (l12) from bacillus stearothermophilus. | the complete amino acid sequence of the ribosomal 'a' protein (bst l12) has been determined from bacillus stearothermophilus. the protein contains 122 amino acids and has a composition of asp4, ans3, thr6, glu20, gln2, pro3, gly9, ala23, val13, met2, ile11, leu8, phe2, lys15, arg1 and a molecular mass of 12737 da. | 1987 | 3609321 |
| effect of plate preincubation on bacillus stearothermophilus disc assay zone diameters. | effect of plate preincubation on the bacillus stearothermophilus disc assay zone size was studied. each of three raw milk samples were subdivided and treated with five levels of us pharmacopeial reference standard penicillin g. freshly prepared, 2.5- and 5-d old seeded disc assay culture plates were preincubated at 64 degrees c for 0 (control), 20, 40, 60, 80, 100, and 120 min prior to placement of discs on the medium surface. plate age did not have a significant effect on the size of the zone d ... | 1987 | 3611471 |
| crystal structure of a deletion mutant of a tyrosyl-trna synthetase complexed with tyrosine. | the crystal structure of a deletion mutant of tyrosyl-trna synthetase from bacillus stearothermophilus has been determined at 2.5 a resolution using molecular replacement techniques. the genetically engineered molecule catalyses the activation of tyrosine with kinetic properties similar to those of the wild-type enzyme but no longer binds trnatyr. it contains 319 residues corresponding to the region of the polypeptide chain for which interpretable electron density is present in crystals of the w ... | 1987 | 3612807 |
| study of the interaction between uncharged yeast trnaphe and elongation factor tu from bacillus stearothermophilis. | proton nmr studies are presented on the interaction of nonaminoacylated yeast trnaphe and elongation factor tu x gtp from bacillus stearothermophilis. from experiments in which transfer of magnetization is observed between proton spins of trna and the protein, it is concluded that complex formation takes place. amino acid residues of the protein come into close contact with the base pair a5u68 and/or u52a62 of the acceptor t psi c limb of the trna molecule. from the line broadening of trna reson ... | 1986 | 3636156 |
| 1-aminocyclopropanephosphonate: time-dependent inactivation of 1-aminocyclopropanecarboxylate deaminase and bacillus stearothermophilus alanine racemase by slow dissociation behavior. | 1-aminocyclopropanephosphonate (acpp) was synthesized, and its effects on the pyridoxal 5'-phosphate linked enzymes 1-aminocyclopropanecarboxylate (acpc) deaminase from pseudomonas sp. acpc and alanine racemase from bacillus stearothermophilus were studied. acpp was found to be a potent inhibitor of both enzymes with km/ki ratios of 500 and 2000, respectively. inhibition for both enzymes was characterized by slow-binding (second-order rate constants less than 150 m-1 s-1) slow-dissociating behav ... | 1987 | 3651390 |
| the isolation of a peptide from the catalytic domain of bacillus stearothermophilus tryptophyl-trna synthetase. the interaction of brown mx-5br with tyrosyl-trna synthetase. | tryptophyl-trna synthetase is irreversibly inactivated by procion brown mx-5br with an apparent dissociation constant (kd) of 8.8 microm and maximum rate of inactivation k3 0.192 s-1. the specificity of the interaction is supported by two previously reported observations. firstly, brown mx-5br inactivation of tryptophyl-trna synthetase is inhibited by substrates, and secondly, the animated derivative of brown mx-5br is a competitive inhibitor of tryptophyl-trna synthetase with a ki of 2 x 10(-4) ... | 1987 | 3663097 |
| the engineering of a more thermally stable lactate dehydrogenase by reduction of the area of a water-accessible hydrophobic surface. | a site-directed mutant of bacillus stearothermophilus lactate dehydrogenase (lactate:nad+ oxidoreductase, ec 1.1.1.27) has been engineered in which the conserved hydrophobic residue isoleucine-250 has been replaced by the more hydrophilic residue asparagine. this isoleucine forms a large part of a water-accessible, hydrophobic surface in the active site of the apo-enzyme which is covered by the b-face of the nicotinamide ring when coenzymes are bound. reduction in the area of this hydrophobic su ... | 1987 | 3663683 |
| heat-stable dna polymerase i large fragment resolves hairpin structure in dna sequencing. | a heat-stable large fragment was obtained by subtilisin digestion of dna polymerase, prepared from bacillus stearothermophilus. the dideoxy sequencing method, combined with the use of m13 vector has proved to be the most powerful one for obtaining the sequences of large genomes. however, the hairpin structure formed along the single-stranded dna template often prevents the dna polymerase from moving on, with the result that no sequence information can be obtained. the heat-stable large fragment ... | 1987 | 3672099 |
| structure and function of l-lactate dehydrogenases from thermophilic and mesophilic bacteria, vi. nucleotide sequences of lactate dehydrogenase genes from the thermophilic bacteria bacillus stearothermophilus, b. caldolyticus and b. caldotenax. | based on the previously determined amino-acid sequence of lactate dehydrogenase from b. stearothermophilus, an oligonucleotide probe was synthesized and used to clone the structural genes for lactate dehydrogenase from b. stearothermophilus, b. caldolyticus and b. caldotenax. the nucleotide sequences of the entire ldh genes from these three thermophilic bacilli were determined by the method of maxam and gilbert. the nucleotide sequence of the ldh gene from b. stearothermophilus is exactly identi ... | 1987 | 3675869 |
| x-ray crystallographic studies of the alanine-specific racemase from bacillus stearothermophilus. overproduction, crystallization, and preliminary characterization. | to facilitate large-scale purification and crystallographic study, we have subcloned the gene for the alanine racemase of bacillus stearothermophilus from picr401 (inagaki, k., tanizawa, k., badet, b., walsh, c. t., tanaka, h., and soda, k. (1986) biochemistry 25, 3268-3274) and overproduced the enzyme in escherichia coli w3110 laciq using the tac promoter of pkk223-3. this system yields alanine racemase as 6% of the bacterial cytosolic protein. purification by a modification of the procedure of ... | 1987 | 3680197 |
| semi-preparative hplc purification of ribosomal proteins from bacillus stearothermophilus and sequence determination of the highly conserved protein s19. | several proteins from the bacillus stearothermophilus 30s ribosomal subunit which could not be isolated by conventional open-column chromatography were purified by high-performance liquid chromatography using a semi-preparative reverse-phase c4 column. protein s19 was purified by this technique and the complete amino acid sequence determined. protein s19 was fragmented and the peptides isolated in picomole quantities were sequenced by an improved manual 4-n,n-dimethylaminoazobenzene-4'-isothiocy ... | 1987 | 3691516 |
| mapping motion in large proteins by single tryptophan probes inserted by site-directed mutagenesis: lactate dehydrogenase. | | 1987 | 3691954 |
| purification and properties of pyruvate kinase from bacillus stearothermophilus. | pyruvate kinase was purified to homogeneity from a moderate thermophile, bacillus stearothermophilus. the molecular weight of the enzyme was found to be 250,000 on gel filtration and 242,000 on sedimentation analysis. the enzyme consisted of four identical subunits of a molecular weight of 62,000-64,000. there were no remarkable differences between the thermophilic enzyme and mesophilic enzymes in amino acid composition, secondary structure, mono- and di-valent cation requirements for activity o ... | 1986 | 3711058 |
| characterization of single crystals of the large ribosomal particles from bacillus stearothermophilus. | single, three-dimensional crystals of the 50 s ribosomal subunit from bacillus stearothermophilus (strain nca) have been characterized using a synchrotron x-ray source. the crystals are orthorhombic with unit cell dimensions: a = 350 a, b = 670 a, c = 905 a, and contain at least one 2-fold screw axis. with cooling to -2 degrees c, the large crystals (1.0 mm x 0.2 mm x 0.1 mm) diffract to 15 to 18 a resolution and are stable in the synchrotron beam for several hours. despite the large cell dimens ... | 1986 | 3712439 |
| comparative thermoresistance of two biological indicators for monitoring steam autoclaves. 3. comparison performed at 121 degrees c in a hospital prevacuum steam sterilizer. | according to pharmacopoea nordica, steam autoclaves should be regularly monitored by a specific swedish preparation of bacillus stearothermophilus spores. if another biological indicator (bi) is used for such a control, it should first be calibrated against the swedish bi (sbi) and the two bis should be equally thermoresistant. attest no. 1262 bi (abi) has previously been shown to be more thermoresistant than the sbi at 134 degrees c, saturated steam. the purpose of the present study was to comp ... | 1986 | 3728026 |
| time-dependent inhibition of bacillus stearothermophilus alanine racemase by (1-aminoethyl)phosphonate isomers by isomerization to noncovalent slowly dissociating enzyme-(1-aminoethyl)phosphonate complexes. | an alanine racemase encoded by a gene from the thermophilic gram-positive bacterium bacillus stearothermophilus is overproduced to 0.3% of the soluble protein when carried on plasmid picr4 in escherichia coli [inagaki, k., tanizawa, k., badet, b., walsh, c. t., tanaka, h., & soda, k. (1986) biochemistry (third paper of four in this issue)]. purification of large quantities (50 mg) of racemase permits study of time-dependent inactivation by d and l isomers of the antibacterial (1-aminoethyl)phosp ... | 1986 | 3730360 |
| protein engineering of homodimeric tyrosyl-trna synthetase to produce active heterodimers. | heterodimers of tyrosyl-trna synthetase from bacillus stearothermophilus have been produced by mutagenesis at the subunit interface. oppositely charged groups have been engineered into the subunits so that they can form a complementary pair. wild-type tyrosyl-trna synthetase is a symmetrical dimer in which the side chains of the 2 phe-164 residues interact at the subunit interface. phe-164 was mutated to asp in tyrosyl-trna synthetase and to lys in a truncated enzyme (des-(321-419)tyrosyl-trna s ... | 1986 | 3733687 |
| [control and use of biological indicators in steam sterilization in the validation of the process and in growth cycles]. | | 1986 | 3741644 |
| dna binding by the bacteriophage spo1-encoded type ii dna-binding protein, transcription factor 1. site-specific binding requires 5-hydroxymethyluracil-containing dna. | the bacteriophage spo1-encoded type ii dna-binding protein, transcription factor 1 (tf1), forms complexes with specific sites in spo1 dna. we have investigated the binding of tf1 to one of its preferred sites in which the normal 5-hydroxymethyluracil (hmura) of spo1 dna has been replaced by thymine and have also investigated the binding of a bacterial type ii dna-binding protein (from bacillus stearothermophilus) to the hmura- and thymine-containing forms of the same dna segment. our results sho ... | 1986 | 3745214 |
| detection of penicillin, cephapirin, and cloxacillin in commingled raw milk by the spot test. | the objective of this study was to compare spot test results with the results of the bacillus stearothermophilus disc assay. commingled raw milk samples were subdivided and spiked with penicillin, cephapirin, or cloxacillin. all subsamples, including unspiked subsamples, were analyzed by the spot test (3 or 9 replicates) and disc assay (9 replicates). mean zone diameter for every subsample was calculated; subsamples having a mean zone diameter greater than or equal to 16 mm were considered posit ... | 1986 | 3745569 |
| structure of a rhamnan from the surface-layer glycoprotein of bacillus stearothermophilus strain nrs 2004/3a. | the structure of a glycan from the surface-layer glycoprotein of bacillus stearothermophilus strain nrs 2004/3a has been studied by 1h- and 13c-n.m.r. spectroscopy. the results indicate the glycan to be a polymer of the trisaccharide repeating-unit ----2)-alpha-l-rhap-(1----2)-alpha-l-rhap-(1----3)-beta-l-++ +rhap-(1----. | 1986 | 3756958 |
| molecular cloning of the gene encoding the valyl-trna synthetase from bacillus stearothermophilus. | the vals gene from the thermophile bacillus stearothermophilus encoding the valyl-trna synthetase has been cloned on a 13.8-kb plasmid. the gene product and its kinetic properties are comparable with those of the native enzyme. | 1986 | 3770480 |
| dipeptide synthesis catalyzed by aminoacyl-trna synthetases from bacillus stearothermophilus. | a new approach to enzymatic peptide synthesis by using aminoacyl-trna synthetase (ars) as a catalyst has been investigated. four arss (asprs, hisrs, leurs and tyrrs) have been purified from a thermophilic bacterium, bacillus stearothermophilus. by using tyrrs as a catalyst, tyrosine and leucinamide were shown to be condensed in the presence of atp to give tyrosylleucinamide. in this manner, all of the arss investigated catalyzed the peptide synthesis reactions. tyrrs did not have strict specific ... | 1986 | 3771102 |
| detergent solubilization of the respiratory nitrate reductase of bacillus stearothermophilus. | four detergents (octyl glucoside, zwittergent, triton x-100, and nonidet p-40) were examined with regard to their efficiency in solubilizing and retaining the activity of the nitrate reductase of bacillus stearothermophilus. at a concentration of between 0.4 and 0.6%, the non-ionic detergent octyl glucoside solubilized only 64% of the membrane proteins. however, about 100% of the nitrate reductase activity was recovered in the soluble fraction. in contrast, the zwitterionic detergent 3-(alkyldim ... | 1986 | 3784923 |
| isolation and identification of two protein-protein crosslinks within the two subunits of bacillus stearothermophilus ribosomes. | bacillus stearothermophilus 30s and 50s ribosomal subunits were isolated and crosslinked with diepoxybutane. the crosslinked proteins were extracted with licl or with 67% acetic acid and purified by a combination of different high performance liquid chromatography techniques. the protein fractions were analysed by two-dimensional and diagonal polyacrylamide gel electrophoresis and by immunological methods. two crosslinked protein pairs, one from the large and one from the small subunit, consisti ... | 1986 | 3790260 |
| temperature-dependent plasmid integration into and excision from the chromosome of bacillus stearothermophilus. | a transformant of bacillus stearothermophilus carrying a recombinant plasmid, plp11 (9.5 mda), on which the penicillinase gene (penp) and kanamycin resistance gene (kan) were located was subjected to mutagenesis, and a mutant plasmid (9.5 mda; penp kan), designated ptra117, was obtained. a transformant of b. stearothermophilus carrying ptra117 could grow at 63 degrees c in medium containing kanamycin, whereas a transformant carrying plp11 could not. although ptra117 was detected as covalently cl ... | 1986 | 3794644 |
| site-directed mutagenesis reveals role of mobile arginine residue in lactate dehydrogenase catalysis. | the binding of substrates to lactate dehydrogenases induces a marked rearrangement of the protein structure in which a 'loop' of polypeptide (residues 98-110) closes over the active site of the enzyme. in this rearrangement, arginine 109 (a basic residue conserved in all known lactate dehydrogenase sequences and in the homologous malate dehydrogenases) moves 0.8 nm from a position in the solvent to one in the active site where its guanidinium group resides within hydrogen bonding distance of bot ... | 1986 | 3796734 |
| the growth of ordered two-dimensional sheets of 70 s ribosomes from bacillus stearothermophilus. | well ordered two-dimensional sheets of intact 70 s ribosomes from bacillus stearothermophilus have been obtained in vitro using salt-alcohol mixtures. these sheets consist of relatively small unit cells with dimensions of 200 +/- 20 a and 400 +/- 30 a. diffraction patterns of electron micrographs of these sheets stained with uranyl acetate contain features to 42 a resolution. | 1986 | 3803568 |
| the isolation and characterization of bacteriophages infecting obligately thermophilic strains of bacillus. | twenty-four thermophilic bacteriophages have been isolated from diverse sources such as compost, soil, silage and rotting straw. although considerable individual host specificity was observed, the phages were able to infect most of the major taxonomic groups of bacillus thermophiles. the phages varied considerably in morphology and size; the phage heads were either cylindrical or polyhedral with tails varying in length between 15 and 500 nm. most of the phages were stable at 50 degrees c for 4-5 ... | 1986 | 3806055 |
| structural and chemical characterization of s-layers of selected strains of bacillus stearothermophilus and desulfotomaculum nigrificans. | the structures, amino acid- and neutral sugar compositions of the crystalline surface layers (s-layers) of four selected strains each of bacillus stearothermophilus and desulfotomaculum nigrificans were compared. among the four strains of each species a remarkable diversity in the molecular weights of the s-layer subunits and in the geometry and constants of the s-layer lattices was apparent. the crystalline arrays included hexagonal (p6), square (p4) and oblique (p2) lattices. in vitro self-ass ... | 1986 | 3813772 |
| characterization of single crystals of the large ribosomal particles from a mutant of bacillus stearothermophilus. | single, three-dimensional crystals of 50 s ribosomal subunits, from a mutant of bacillus stearothermophilus that lacks the protein l11, have been characterized using a synchrotron x-ray source. the crystals of the mutated particles grow under the same conditions and are isomorphous to those of the wild type of the same bacteria. they are orthorhombic, contain at least one 2-fold screw axis, and have unit cell dimensions of a = 350(+/- 10) a, b = 670(+/- 10) a, and c = 910(+/- 10) a. they diffrac ... | 1986 | 3820303 |
| hydrogen bonding and biological specificity analysed by protein engineering. | the role of complementary hydrogen bonding as a determinant of biological specificity has been examined by protein engineering of the tyrosyl-trna synthetase. deletion of a side chain between enzyme and substrate to leave an unpaired, uncharged hydrogen-bond donor or acceptor weakens binding energy by only 0.5-1.5 kcal mol-1. but the presence of an unpaired and charged donor or acceptor weakens binding by a further approximately 3 kcal mol-1. | 1985 | 3845322 |
| two-step purification of tryptophan-accepting trna from bacillus stearothermophilus. | tryptophan-accepting trna has been purified essentially to homogeneity from bacillus stearothermophilus. crude trna was chromatographed first on benzoylated deae-cellulose and then on sepharose 4b with reverse salt gradient elution. the product has tryptophan acceptor activity in excess of 2 nmol [14c]tryptophan per a260 unit. this procedure avoids costly aminoacylation, a step characteristic of other one- and two-step procedures. in two separate purifications 7 and 11 mg of trnatrp were prepare ... | 1985 | 3854274 |
| redesigning enzymes by site-directed mutagenesis. | the systematic alteration of protein structure has now become possible with genetic engineering. recent developments in techniques for the chemical synthesis of dna fragments and in recombinant dna technology have enabled the facile modification of proteins by highly specific mutagenesis of their genes. enzymes with novel properties may be produced in large quantities from the mutant genes. kinetic analysis of the mutant enzymes can be combined with high-resolution structural data from protein x ... | 1985 | 3860366 |
| transition-state stabilization in the mechanism of tyrosyl-trna synthetase revealed by protein engineering. | the principal catalytic factor in the activation of tyrosine by the tyrosyl-trna synthetase is found to be improved binding of atp in the transition state. the activation reaction involves the attack of the tyrosyl carboxylate on the alpha-phosphate group of atp to generate a pentacoordinate transition state. model building of this complex located a binding site for the gamma-phosphate group of atp, consisting of hydrogen bonds with the side chains of thr-40 and his-45. removal of these groups b ... | 1985 | 3865201 |
| complete nucleotide sequence of a thermophilic alpha-amylase gene: homology between prokaryotic and eukaryotic alpha-amylases at the active sites. | the nucleotide sequence of a thermophilic, liquefying alpha-amylase gene cloned from b. stearothermophilus was determined. the nh2-terminal amino acid sequence analysis of the b. stearothermophilus alpha-amylase confirmed that the reading frame of the gene consisted of 1,644 base pairs (548 amino acids). the b. stearothermophilus alpha-amylase had a signal sequence of 34 amino acids, which was cleaved at exactly the same site in e. coli. the mature enzyme contained two cysteine residues, which m ... | 1985 | 3876333 |
| cloning and expression of the alpha-amylase gene from bacillus stearothermophilus in several staphylococcal species. | the plasmid-coded alpha-amylase gene of bacillus stearothermophilus (amy) was cloned in staphylococcus carnosus using plasmid pca43 as a vector. the amy gene was located on a 5.4-kb hindiii dna fragment of the hybrid plasmid pamy7. when transformed into other staphylococcal species, plasmid pamy7 exhibited marked differences in the production of alpha-amylase (alpha amy). most active for heterospecific alpha amy production was staphylococcus aureus. in its culture supernatant nearly half as much ... | 1985 | 3876968 |
| electron microscopy of the secondary structure in partially denatured rrnas of escherichia coli and bacillus stearothermophilus. | partially denatured 16s and 23s rrnas from the thermophile bacillus stearothermophilus show characteristic loop patterns when observed by electron microscopy. the patterns are very similar to those seen in rrnas from escherichia coli. at least 2 of 4 most stable interactions in 16s rrna and 8 of 12 interactions in 23s rrna are in common for the two species. these interactions correspond well to features of secondary structure in models inferred for rrna from phylogenetic sequence comparisons and ... | 1985 | 3882675 |
| comparison of eubacterial and eukaryotic 5s rna structures: a chemical modification study. | the 5s rnas from bacillus stearothermophilus and saccharomyces cerevisiae were probed by nucleotide-specific reagents, with a view to compare and contrast their higher order structures. the progressive unfolding of the rnas during heating, in the presence and absence of magnesium, was monitored. evidence was provided for the double-helical segments which occur in the secondary structural models of both rnas. the results also placed constraints on the possible structuring of the remainder of the ... | 1985 | 3884039 |
| on a sequence similarity between ribosomal protein s5 and dna binding protein ii. | | 1985 | 3884370 |
| a note on an antimicrobial agent in disc paper and a modified assay procedure for detecting antimicrobials in milk. | commercially available disc paper used for antibiotic sensitivity tests was shown to contain an unidentified, water-soluble agent inhibitory to bacillus stearothermophilus which gave false positive results in an antimicrobial assay for milk. the agent however, was found to be inactive against a range of common pathogenic bacteria. it was easily removed by washing the discs in distilled water before use. | 1985 | 3888940 |
| evolutionary conservation of protein binding sites on high-molecular-mass ribosomal rna. | | 1985 | 3899176 |
| two-dimensional polyacrylamide gel electrophoresis of ribosomal proteins in the nanogram range. | a two-dimensional gel electrophoresis system to identify and check the purity of ribosomal proteins from different organisms with nanogram quantities is described. this procedure combines the method of geyl et al. for the separation of ribosomal proteins of escherichia coli, and the microscale electrophoresis system for proteins described by neuhoff and poehling, with several modifications. the first gel dimension is carried out in capillaries and the second in the form of slab gels, both are ru ... | 1985 | 3907663 |
| characteristic views of e. coli and b. stearothermophilus 30s ribosomal subunits in the electron microscope. | large sets of electron microscopic images of the 30s ribosomal subunits of bacillus stearothermophilus (914 molecules) and escherichia coli (422 molecules) were analysed with image processing techniques. using computer alignment and a new multivariate statistical classification scheme, three predominant views of the subunit were found for both species. these views, which together account for approximately 90% of the population of images, were determined to a reproducible resolution of up to 1.7 ... | 1985 | 3908096 |
| the primary structure of ribosomal protein l2 from bacillus stearothermophilus. | the complete amino acid sequence of ribosomal protein l2 from the moderate thermophile bacillus stearothermophilus has been determined. this has been achieved by the sequence analysis of peptides derived by enzymatic digestion with staphylococcus aureus protease, trypsin and chymotrypsin, as well as by chemical cleavage with o-iodosobenzoic acid. the protein contains 275 amino acid residues and has a calculated molecular mass of 30201 da. comparison of this sequence with sequences of the corresp ... | 1985 | 3908098 |
| engineering of tyrosyl trna synthetase. | the gene encoding the enzyme tyrosyl trna synthetase from bacillus stearothermophilus has been systematically altered using synthetic oligonucleotides as mutagens. the construction of mutations has been facilitated by using strains of bacteria defective in mismatch repair and also by utilising a genetic marker in the m13 strain (such as an amber mutation, or an ecok or ecob site) which allows selection for the progeny of m13 replication derived from the minus (mutagenized) strand. several mutati ... | 1985 | 3910110 |
| purification and characterization of heat-stable proteases from bacillus stearothermophilus rm-67. | the extracellular proteases of bacillus stearothermophilus rm-67 were purified by ammonium sulfate fractionation (40 to 70% saturation), gel filtration through sephadex g-100, and diethylaminoethyl-sephadex a-50 ion-exchange chromatography. gel filtration resulted in separation of the enzyme preparation into one minor (protease i) and one major (protease ii) peak. the three-step purification scheme resulted in 39.5-fold purification and an overall recovery of 8.1% of protease i and 87.8-fold pur ... | 1985 | 3912410 |
| isolation and characterization of antibiotic-resistance plasmids in thermophilic bacilli. | four antibiotic-resistance plasmids isolated from thermophilic bacilli were characterized in detail. three tetracycline-resistance (tc1) plasmids were designated as ptht9 (7.7 kilobases (kb], ptht15 (4.5 kb) and ptht22 (8.4 kb). from the results of restriction endonuclease analysis and the subsequent southern hybridization, these were found to possess extensive genetic homology in the regions that include the replication origin and the tcr gene. detailed restriction maps of the smallest tcr plas ... | 1985 | 3924385 |
| nucleotide sequence of the bacillus stearothermophilus alpha-amylase gene. | the nucleotide sequence of the bacillus stearothermophilus alpha-amylase gene and its flanking regions was determined. an open reading frame was found, comprising a total of 1,647 base pairs (549 amino acids) and starting from a gug codon as methionine. it was shown by nh2-terminal amino acid sequence analysis that the extracellular amylase consisted of 515 amino acid residues, which corresponded to a molecular weight of 58,779. thus the nh2-terminal portion of the gene encodes 34 amino acid res ... | 1985 | 3924897 |
| new perspectives on bacterial ferredoxin evolution. | recent evidence indicates that a gene transposition event occurred during the evolution of the bacterial ferredoxins subsequent to the ancestral intrasequence gene duplication. in light of this new information, the relationships among the bacterial ferredoxins were reexamined and an evolutionary tree consistent with this new understanding was derived. the bacterial ferredoxins can be divided into several groups based on their sequence properties; these include the clostridial-type ferredoxins, t ... | 1985 | 3932661 |
| spore heat resistance and specific mineralization. | spores of bacillus megaterium atcc 19213, bacillus subtilis niger and bacillus stearothermophilus atcc 7953 were converted to fully demineralized, but viable, h forms by controlled acid titration. h forms were more heat sensitive than were native forms, but z values were greater for killing of h spores than those for native spores. therefore, the differences in heat sensitivity between native and h forms decreased with increasing killing temperature. the increase in heat sensitivity associated w ... | 1985 | 3937495 |
| a method for calculating thermal sterilization conditions based upon process parametrics. | | 1985 | 3937893 |
| high-multiplicity spin states of 2[4fe-4se]+ clostridial ferredoxins. | the electron paramagnetic resonance (epr) spectra of the reduced selenium-substituted 2-[4fe-4se]+ ferredoxins from three bacteria of the clostridium genus display low-field signals at g = 5.17, g = 10.11, and g = 12.76. the positions, shapes, and temperature dependencies of these signals have allowed their assignments to the three excited states of an s = 7/2 spin multiplet, the fundamental state of which is observed as unusual features in low-temperature (t less than or equal to 20 k) mössbaue ... | 1986 | 3955006 |
| a model of synthetase/transfer rna interaction as deduced by protein engineering. | the recognition of transfer-rna by their cognate aminoacyl-trna synthetases is the crucial step in the translation of the genetic code. in order to construct a structural model of the complex between the tyrosyl-trna synthetase (tyrts) from bacillus stearothermophilus and trnatyr, 40 basic residues at the surface of the tyrts dimer have been mutated by site-directed mutagenesis and heterodimers created in vitro by recombining subunits derived from different mutants. as reported here a cluster of ... | 1986 | 3960121 |
| identity of the metal ligands in the manganese- and iron-containing superoxide dismutases. | alignment of the amino acid sequence of peptides obtained following digestion of photobacterium leiognathi iron superoxide dismutase with the known sequence of bacillus stearothermophilus manganese superoxide dismutase shows that the residues found to form ligands to the manganese are conserved in the iron enzyme. this indicates that the metal ligands in both proteins are identical. | 1985 | 3967757 |
| changes in the state of subunit association of lactate dehydrogenase from bacillus stearothermophilus. | time-resolved measurements of the fluorescence anisotropy of an extrinsic dye-group attached to lactate dehydrogenase from b. stearothermophilus revealed that the rotational correlation time of the enzyme at low concentrations is 55 ns, while at high enzyme concentrations or in the presence of fructose 1,6-bisphosphate (fru-1,6-p2) the correlation time increases to 95 ns. these correlation times are consistent with a change in mr from 85 000 +/- 12 000 (dimer) to 150 000 +/- 22 000 (tetramer) an ... | 1985 | 3986214 |
| protoplast dehydration correlated with heat resistance of bacterial spores. | water content of the protoplast in situ within the fully hydrated dormant bacterial spore was quantified by use of a spore in which the complex of coat and outer (pericortex) membrane was genetically defective or chemically removed, as evidenced by susceptibility of the cortex to lysozyme and by permeability of the periprotoplast integument to glucose. water content was determined by equilibrium permeability measurement with 3h-labeled water (confirmed by gravimetric measurement) for the entire ... | 1985 | 3988704 |
| the effect of growth temperature on the thermotropic behavior of the membranes of a thermophilic bacillus. composition-structure-function relationships. | the following study was carried out with the aim of widening our understanding of the thermoadaptive mechanisms of the membrane of thermophiles, using bacillus stearothermophilus var. nondiastaticus as test-organism. the phospholipids and their acyl chain composition of this bacillus studied in relation to the physical properties of its membrane from bacteria grown at various temperatures. phospholipids account for 68-75 weight% of the total lipid in cells grown at 45, 55 or 65 degrees c. phosph ... | 1985 | 3995029 |
| comparative thermoresistance of two biological indicators for monitoring steam autoclaves. 2. comparison performed at 134 degrees c in a hospital prevacuum steam sterilizer. | the thermoresistance of various lots of two biological indicators (bis) for steam sterilization control, a scandinavian bi (sbi) and the attest bi (abi), were compared during sterilization cycles in a hospital prevacuum (pressure-pulsing) steam autoclave at 134 degrees c, saturated steam. abi no. 1242, abi no. 1262 (its replacement) and incremental heat exposure times between 0 s and 180 s were used. the intrachamber temperature and pressure were continuously measured and monitored throughout th ... | 1985 | 4013742 |
| the complete primary structure of ribosomal proteins l1, l14, l15, l23, l24 and l29 from bacillus stearothermophilus. | the amino acid sequences of ribosomal proteins l1, l14, l15, l23, l24 and l29 from bacillus stearothermophilus have been completely determined. this has been achieved by sequence analyses of peptides derived from enzymatic digestions of the proteins with trypsin, chymotrypsin, pepsin, staphylococcus aureus protease, and armillaria mellea protease as well as by chemical cleavage with hydroxylamine and cyanogen bromide. based on the primary structures of the six proteins, their secondary structure ... | 1985 | 4018095 |
| artificial and natural thermostabilization of subunit enzymes. do they have similar mechanism? | rabbit skeletal muscle glyceraldehyde-3-phosphate dehydrogenase was stabilized by intramolecular intersubunit crosslinking with diimidoesters. half-inactivation temperature for optimal cross-linker-treated enzyme preparation increased by 11 degrees c. stabilization effect correlated with the content of crosslinked fractions in enzyme preparation, as proved by sds gel-electrophoresis. it is proposed that artificial crosslinks stabilize the enzyme in a similar fashion to salt bridges in the thermo ... | 1985 | 4029485 |
| thermophilic nad-dependent glutamate dehydrogenase from bacillus stearothermophilus. | a rapid purification procedure for glutamate dehydrogenase (gdh) from bacillus stearothermophilus var calidolactis was developed. the homogeneous enzyme with a total molecular weight of approximately 240,000 daltons, contained 6 identical subunits. no high molecular weight form of gdh present in crude extracts was found after elution of the enzyme from a 5'amp-sepharose column with 4 m urea. the purified enzyme functions in both directions i.e. amination and deamination and is strictly specific ... | 1985 | 4038315 |
| stability and self-assembly of the s-layer protein of the cell wall of bacillus stearothermophilus. | the surface layer of the cell envelope of bacillus stearothermophilus consists of a regular array of protein subunits. as shown by dodecyl sulfate polyacrylamide gel-electrophoresis and ultracentrifugation, the fully solubilized s-layer protein represents a homogeneous entity with a subunit molecular mass of 115 +/- 5 kda. solubilization of the protein may be accomplished at acid ph, or using high concentrations of urea or guanidine x hcl. it is accompanied by (partial) denaturation, thus interf ... | 1985 | 4041240 |
| purification and properties of glucose-6-phosphate dehydrogenase from bacillus stearothermophilus. | glucose-6-phosphate dehydrogenase (d-glucose-6-phosphate: nadp+ 1-oxidoreductase, ec 1.1.1.49) from the thermophilic bacteria, bacillus stearothermophilus, was purified and its properties were examined. the enzyme was shown to consist of four identical subunits, each of about mr 50,000. this enzyme utilized both nadp+ and nad+ as cofactors, and the maximum velocity for both cofactors was similar. however, the km values were quite different from each other, being 0.016 and 1.64 mm for nadp+ and n ... | 1985 | 4055554 |
| probing histidine-substrate interactions in tyrosyl-trna synthetase using asparagine and glutamine replacements. | we have analyzed the interactions of a histidine residue with a substrate using site-directed mutagenesis. previous studies on tyrosyl-trna synthetase from bacillus stearothermophilus have shown that a histidine residue (his-48) makes an interaction with atp, which is improved on mutating thr-51----pro-51. we find on replacing his-48 in wild-type enzyme with either asparagine or glutamine that asn-48 is equally as good as his-48 but his-48----gln-48 leads to a far lower activity. the side chain ... | 1985 | 4074680 |
| location of the binding region for 23s ribosomal rna on ribosomal protein l2 from bacillus stearothermophilus. | tryptic and chymotryptic digestions of a complex between ribosomal protein l2 and the 23s rna of bacillus stearothermophilus produced two protected fragments (designated tf and cf). these protein fragments remained tightly bound to the rna and were able to rebind specifically to 23s rna. amino acid sequence analysis of the isolated fragments revealed that both fragments tf and cf originated from the central half of the protein l2 (positions 60-206 and 58-201) with molecular masses of 15918 da an ... | 1985 | 4076177 |
| diffusion test for the detection of streptomycin residues in milk. | | 1985 | 4078118 |
| reversible dissociation of dimeric tyrosyl-trna synthetase by mutagenesis at the subunit interface. | dimeric tyrosyl-trna synthetase from bacillus stearothermophilus exhibits half-of-the-sites reactivity and negative cooperativity in binding of tyrosine. protein engineering has been applied to the enzyme to determine whether it can be reversibly dissociated into monomers and if the monomers are active. the target for mutation is the residue phe-164. the side chain of phe-164 in one subunit interacts with its symmetry-related partner in the other. mutation of phe-164----asp-164 gives a mutant [t ... | 1985 | 4084496 |
| determination of the heat resistance of spores using a solid heating block system. | a simple and accurate technique for the determination of the heat resistance of spores is described. the technique combines a modified capillary tube method with a solid heating block. the come-up time of spore suspensions was found to be short and simple and accurate technique is suggested for the correction of the come-up times. experimental results are presented for the destruction of spores of bacillus stearothermophilus at 120 degrees which indicates the accuracy and reproducibility of the ... | 1985 | 4086408 |
| the release of dipicolinic acid during heating and its relation to the heat destruction of bacillus stearothermophilus spores. | the rates of dipicolinic acid (dpa) release and the rates of death were studied for spores of five strains of bacillus stearothermophilus. it was observed that a highly significant relationship exists between the rate of dpa release and rate of spore death for the four out of five strains tested and for all test temperatures. at 115 degrees c the rate of dpa release was found to be faster than the rate of death, equal at 120 degrees c and slower at 125 degrees c. the role of dpa in heat resistan ... | 1985 | 4086410 |
| ultraviolet spectrophotometric method for determination of cholinesterase activity with acetylcholine as a substrate. | an enzymatic method for the determination of serum cholinesterase (che) activity is described. the method is based on the liberation of acetate from acetylcholine as a substrate by che and the conversion of the acetate to acetylphosphate and adp in the presence of atp by acetate kinase. the produced adp is coupled with pyruvate kinase and lactate dehydrogenase in the presence of phosphoenolpyruvate and nadh. the amount of nadh consumed is determined by absorbance at 340 nm. the reaction proceeds ... | 1985 | 4093350 |
| [estimation of the phylogenetic distance between prokaryotic species through the use of ribosomal proteins]. | | 1974 | 4218537 |
| [determination of kinetic in activation data on vapor-heated, dried spores in a suitable thermoresistometer]. | | 1974 | 4446694 |
| [sterilization of solutions with reference to the decay kinetics of microorganisms and substrate destruction]. | | 1974 | 4460007 |
| the thermal resistance of bacillus stearothermophilus spores. the effects of temperature and ph of the heating medium. | | 1974 | 4462138 |
| evolution of ribosomal proteins. | | 1974 | 4619938 |
| the fine-structural properties of the amino-terminal region of the 50s ribosomal "a" protein from bacillus stearothermophilus. | | 1974 | 4619940 |
| location of protein s4 on the small ribosomal subunit of e. coli and b. stearothermophilus with protein- and hapten-specific antibodies. | in spite of considerable effort there is still serious disagreement in the literature about the question of whether epitopes of ribosomal protein s4 are accessible for antibody binding on the intact small ribosomal subunit. we have attempted to resolve this issue using three independent approaches: (i) a re-investigation of the exposure and the location of epitopes of ribosomal protein s4 on the surface of the 30s subunit and 30s core particles of the e. coli ribosome, including rigorous control ... | 1984 | 6083434 |
| electron-paramagnetic-resonance studies of manganese(ii) complexes with elongation factor tu from bacillus stearothermophilus. observation of a gtp hydrolysis intermediate state complex. | changes in the coordination of mn2+ to nucleotide, water and protein at the active site of elongation factor tu (ef-tu) have been studied by electron paramagnetic resonance (epr) spectroscopy. from the time dependence of the mn2+ spectrum after addition of gtp to ef-tu x mn, it was apparent that three complexes with different epr linewidths could be detected. using additional information from the kinetics of 32pi production and release from ef-tu x mn x [gamma-32p]gtp these were identified as ef ... | 1984 | 6086334 |
| enzymatic and nucleotide sequence studies of a kanamycin-inactivating enzyme encoded by a plasmid from thermophilic bacilli in comparison with that encoded by plasmid pub110. | the product of a kanamycin resistance gene encoded by plasmid ptb913 isolated from a thermophilic bacillus was identified as a kanamycin nucleotidyltransferase which is similar to that encoded by plasmid pub110 from a mesophile, staphylococcus aureus. the enzyme encoded by ptb913 was more thermostable than that encoded by pub110. in view of a close resemblance of restriction endonuclease cleavage maps around the bglii site in the structural genes of both enzymes, ca. 1,200 base pairs were sequen ... | 1984 | 6090428 |
| two replication determinants of an antibiotic-resistance plasmid, ptb19, from a thermophilic bacillus. | two different replication determinants were found on an antibiotic resistance plasmid, ptb19, from a thermophilic bacillus. one replication determinant (designated repa) was functional only in bacillus subtilis, whereas the other (designated repb) functioned in both b. subtilis and bacillus stearothermophilus. a deletion plasmid, ptb90, carrying the repb derived from ptb19 coincidentally contained the specific 1.0 mdal ecori fragment of a cryptic plasmid pbso2 from b. stearothermophilus. the pre ... | 1984 | 6090582 |
| continuous atp regeneration process with stable acetate kinase. | heat-stable acetate kinase (ak) from bacillus stearothermophilus was successfully immobilized covalently to sepharose resin by several conventional methods including carbodiimide, hydroxysuccinimide, cyanogen bromide, and glutaraldehyde and also by a new method which utilizes a bifunctional adp derivative as a spacer. the latter method gave a higher yield in terms of enzyme activity than the conventional methods. the properties and kinetics of the immobilized ak were studied batchwise and in a c ... | 1984 | 6092323 |
| nucleotide sequence of the triose phosphate isomerase gene of escherichia coli. | we report here the complete nucleotide sequence of the e. coli triose phosphate isomerase gene. the gene encodes a polypeptide of 255 amino acids which is approximately 46% homologous to eukaryotic triose phosphate isomerases, and approximately 38% homologous to the enzyme from a thermophilic bacterium, bacillus stearothermophilus. the nucleotide sequence is 55% homologous to that of the corresponding gene in the yeast saccharomyces cerevisiae. to our knowledge, this is the first report of the s ... | 1984 | 6092857 |
| complete nucleotide sequence of a 23s ribosomal rna gene from bacillus stearothermophilus. | a 23s ribosomal (rrna) gene from bacillus stearothermophilus has been cloned in pbr322, and its nucleotide sequence determined. the corresponding mature 23s rrna is predicted to contain 2928 nucleotides. we compare the primary and secondary structures of 23s rrna from escherichia coli and b. stearothermophilus, and discuss their potential contributions to thermal stability. | 1984 | 6096097 |
| reversible acid dissociation of thermostable inorganic pyrophosphatase from bacillus stearothermophilus. | | 1980 | 6101557 |
| primary structure of triosephosphate isomerase from bacillus stearothermophilus. | 1. triosephosphate isomerase from bacillus stearothermophilus is a dimeric enzyme comprising two chemically identical polypeptide chains. 2. the nearly complete amino acid sequence of the subunit polypeptide chain has been established from sequences of tryptic, chymotryptic and lysine-blocked tyrptic fragments of s-[2-14c]carboxymethylated enzyme. overlaps not established by experimental data have been provisionally established from considerations of sequence homology with previously established ... | 1980 | 6105959 |
| evaluation of a prototype micro-electronic autoclave cycle integrator. | instability and non-uniformity of spore preparations and the non-conformity of chemical indicators to the temperature coefficients of spore inactivation are problems associated with current methods of autoclave cycle monitoring. a prototype micro-electronic instrument, which largely overcomes these problems, is described. it monitors autoclave cycles in terms of the integral f0 and nabla functions. its thermometric and integrating accuracy is demonstrated. a discussion of the problems associated ... | 1981 | 6111601 |
| phosphofructokinase: structure and control. | phosphofructokinase from bacillus stearothermophilus shows cooperative kinetics with respect to the substrate fructose-6-phosphate (f6p), allosteric activation by adp, and inhibition by phosphoenolpyruvate. the crystal structure of the active conformation of the enzyme has been solved to 2.4 a resolution, and three ligand-binding sites have been located. two of these form the active site and bind the substrates f6p and atp. the third site binds both allosteric activator and inhibitor. the comple ... | 1981 | 6115424 |
| evaluation of glycine as an inactivator of glutaraldehyde. | glycine was evaluated as an inactivator of the sporicidal activity of glutaraldehyde. spores from glucose depleted cultures of bacillus stearothermophilus grown in a chemically defined medium were used. when glycine is used as an inactivator of glutaraldehyde, it lowers the ph value of the solution. glycine 1% failed to inactivate 0.5% or higher concentrations of alkaline glutaraldehyde in sporicidal studies. if viable counts cannot be performed within the first hour after inactivation, the con ... | 1982 | 6124593 |
| a comparison of heat resistance in commercially available bacillus stearothermophilus spore preparations used for monitoring steam sterilization. | | 1982 | 6124601 |
| composition and properties of trypsin-cleaved elongation factor tu. | native elongation factor tu from escherichia coli, ef-tu, is initially attacked by trypsin at three adjacent sites in the primary structure. these are arginine-44, arginine-58, and lysine-56. the rates of hydrolysis at the two arginine residues are about the same but that at the lysine residue is much slower. the products of the tryptic digestion have been analysed by edman degradation and polyacrylamide gel electrophoresis. the peptide from alanine-45 to arginine-58 is eventually excised and do ... | 1980 | 6157532 |
| differential staining of the subunits of dna-dependent rna polymerase. | | 1980 | 6160057 |
| the production of alpha-amylase in batch and chemostat culture by bacillus stearothermophilus. | the production of alpha-amylase (alpha-1,4-glucan 4-glucanohydrolase; ec 3.2.1.1) by a strain of bacillus stearothermophilus isolated from leaf litter was investigated in a tryptone-maltose medium at 55 degrees c in batch and chemostat culture. amylase production was growth-limited and restricted to the exponential phase in batch culture. the enzyme yield was reduced by 40% when the culture ph was maintained at ph 7.2. amylase production in chemostat culture was influenced by the growth rate thr ... | 1980 | 6160812 |
| stringent response of bacillus stearothermophilus: evidence for the existence of two distinct guanosine 3',5'-polyphosphate synthetases. | bacillus stearothermophilus reacted to pseudomonic acid-induced inhibition of isoleucine-transfer ribonucleic acid (rna) acylation and to energy downshift caused by alpha-methylglucoside addition with accumulation of guanosine 3',5'-polyphosphates [(p)ppgpp] and restriction of rna synthesis. in vitro studies indicated that (p)ppgpp was synthesized by two different enzymes. one enzyme, (p)ppgpp synthetase i, was present in the ribosomal fraction, required the addition of a ribosome-messenger rna- ... | 1981 | 6161916 |
| the stringent response to unacylated trna, energy-and temperature-downshift in bacillus stearothermophilus. | the response of the thermophile bacillus stearothermophilus to inhibition of trna acylation, energy starvation and temperature downshift was characterized. we found that b. stearothermophilus, like other prokaryotic organisms, reacts with the so-called stringent response, which includes the accumulation of the unusual nucleotides guanosine 3',5' bis (diphosphate) [ppgpp] and guanosine 3'-diphosphate, 5'-tri-phosphate [pppgpp] and concomitantly the reduction of rna synthesis and growth rate. the ... | 1981 | 6164348 |
| reconstitution of biologically active 50s ribosomal subunits with artificial 5s rna molecules carrying disturbances in the base pairing within the molecular stalk. | bacillus stearothermophilus 50s ribosomal subunits were reconstituted in vitro using artificial 5s rna molecules constructed by combining parts of major and minor type (raué et al. (1976) europ. j. biochem. 68, 169-176) b. licheniformis 5s rna. the artificial 5s rna molecules carry defined disturbances (a.c juxtapositions and extra g.u pairs) in the base pairing between the 5'- and 3'-terminal sequences of the molecule (the molecular stalk region). the biological activity of the reconstituted su ... | 1981 | 6164987 |
| fluorescence modification of escherichia coli 5s rna. | reaction of 5s rna with chlorocetaldehyde leads to the conversion of unpaired adenines to the fluorescent 1,n6-etheno-adenine derivatives. up to 16 of the 23 adenines in free 5s rna can be modified, the fastest reacting are a29, a34, a57-59. partial modification of adenines in this area results in a 20% reduction in the efficiency of 5s rna incorporation into 50s subunits during reconstitution and a 15% reduction in the activity of these subunits in peptide synthesis. fluorescence from 1,n6-ethe ... | 1981 | 6169004 |
| tests on a low temperature steam and formaldehyde autoclave: the miniclave 80. | | 1981 | 6174581 |