Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| 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 |
| buffering capacity of bacilli that grow at different ph ranges. | cytoplasmic buffering capacities and buffering by whole cells were examined in six bacterial species: bacillus acidocaldarius, bacillus stearothermophilus, escherichia coli, bacillus subtilis, bacillus alcalophilus, and bacillus firmus rab. acid-base titrations were conducted on whole cells and cells permeabilized with triton x-100 or n-butanol. in all of the species examined, the buffering capacity of intact cells was generally a significant proportion of the total buffering capacity, but the m ... | 1985 | 3886633 |
| 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 |
| 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 |
| 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 |
| use of microbial beta-lactamase to destroy penicillin added to milk. | a simple method is described for destruction of penicillin residues in bulk milk to an undetectable amount (less than .003 u/ml) with commercially available crude beta-lactamase enzyme. milk containing .1 or .5 u/ml penicillin g was treated with .01 to 1.0 mu/ml of beta-lactamase (bacillus cereus) for up to 96 h. the bacillus stearothermophilus var. calidolactis assay was used to quantify penicillin in milk between .003 to 1.0 u/ml. the .5 u/ml of penicillin g was reduced to an undetectable amou ... | 1985 | 3930584 |
| 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 |
| the immobilization of enzymes and cells of bacillus stearothermophilus onto poly(maleic anhydride/styrene)-co-polyethylene and poly(maleic anhydride/vinyl acetate)-co-polyethylene. | the graft copolymer, poly(maleic anhydride/styrene)-co-polyethylene was prepared. the copolymer immobilized bovine serum albumin (bsa), but the amount coupled appeared to be effected by the amount of styrene in the graft copolymer, temperature, and ph of the coupling medium. competition existed between hydrolysis of the grafted anhydride groups and the protein. a graft copolymer with 66% add-on immobilized 4.5 mg/glucose oxidase/g copolymer, 4.6 mg alkaline phosphates/g copolymer and 0.2 mg cell ... | 1985 | 18553712 |
| effects of temperature and dilution rate on the copy number of recombinant plasmid in continuous culture of bacillus stearothermophilus (plp11). | the penicillinase yield y(p/x) (u/g cell) of a transformant bacillus stearothermophilus cu21 (plp11) in continuous culture of lgp broth at 44, 47.5, and 50 degrees c, respectively, depended not only on temperature, theta ( degrees c), but also on dilution rate, d (h(-1)), in a peculiar fashion that could not have been realized if the product were from the gene on the chromosome rather than the plasmid. the gene dosage effect could account for the unusual dependence of y(p/x) on theta and d, beca ... | 1985 | 18553728 |
| high production of thermostable beta-galactosidase of bacillus stearothermophilus in bacillus subtilis. | by cloning the beta-galactosidase gene of bacillus stearothermophilus iam11001 (atcc 8005) into bacillus subtilis, enzyme production was enhanced 50 times. beta-galactosidase could be purified to 80% homogeneity by incubating the cell extract of b. subtilis at 70 degrees c for 15 min, followed by centrifugation to remove the denatured proteins. because of its heat stability and ease of production, beta-galactosidase is suitable for application in industrial processes. | 1985 | 16346825 |
| culture conditions for production of thermostable amylase by bacillus stearothermophilus. | bacillus stearothermophilus grew better on complex and semisynthetic medium than on synthetic medium supplemented with amino acids. amylase production on the complex medium containing beef extract or corn steep liquor was higher than on semisynthetic medium containing peptone (0.4%). the synthetic medium, however, did not provide a good yield of extracellular amylase. among the carbohydrates which favored the production of amylase are, in order starch > dextrin > glycogen > cellobiose > maltohex ... | 1986 | 16347107 |
| immobilization of bsa, enzymes and cells of bacillus stearothermophilus onto cellulose, polygalacturonic acid and starch based graft copolymers containing maleic anhydride. | poly(maleic anhydride styrene) graft copolymers of cellulose, pectin polygalacturonic acid salt, calcium polygalacturonate, and starch were prepared and used to immobilize proteins. the cellulose grafts coupled quite appreciable quantities of acid phosphatase, glucose oxidase, and trypsin. however, the general retention of activity was somewhat disappointing. further investigation with acid phosphatase showed that the amount of enzyme immobilized increased as the amount of anhydride in the graft ... | 1986 | 18553841 |
| stabilization of kappa-carrageenan gel with polymeric amines: use of immobilized cells as biocatalysts at elevated temperatures. | spherical beads of kappa-carrageenan containing entrapped cells were prepared in a two-step process. first, the beads were formed by dispersing a warm carrageenan cell suspension into stirring oil. after cooling (gelation) the beads were cured by treatment with amines. ten amines of various sizes and structures were tested. we evaluated the mechanical strength and the applicability of aminetreated gels as immobilization matrices. the results of critical compression tests indicate that linear and ... | 1986 | 18561217 |
| 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 |
| 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 |
| 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 |
| 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 effect of growth temperature on esterase patterns in psychrotrophic bacillus species and other gram-positive genera. | the growth temperature of bacillus stearothermophilus was previously reported to affect the esterase band pattern obtained after gel electrophoresis. in this study gel electrophoresis of esterases was done on a group of antarctic bacterial strains cultivated at different temperatures to investigate whether band shift due to a change in growth temperature was a general phenomenon or limited only to select groups. most strains studied were in the bacillus genus. standard strains of known bacillus ... | 1986 | 3813524 |
| 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 |
| structural homology between elongation factors ef-tu from bacillus stearothermophilus and escherichia coli in the binding site for aminoacyl-trna. | elongation factor ef-tu (mr approximately equal to 50 000) and elongation factor ef-g (mr approximately equal to 78 000) were isolated from bacillus stearothermophilus in a homogeneous form. the ability of ef-tu to participate in protein synthesis is rapidly inactivated by n-tosyl-l-phenyl-alanylchloromethane (tos-phech2cl). ef-tu x gtp is more susceptible to the inhibition by tos-phech2cl than is ef-tu x gdp. tos-phech2cl forms a covalent equimolar complex with the factor by reacting with a cys ... | 1986 | 3510872 |
| genetic analysis of bacillus stearothermophilus by protoplast fusion. | efficient and reliable protoplasting, regeneration, and fusion techniques were established for the prototrophic strain bacillus stearothermophilus nub36. auxotrophic mutants were isolated, and protoplast fusion was used to construct isogenic mutant strains and for chromosomal mapping. markers were mapped using two-, three-, and four-factor crosses. the order of the markers was hom-1-thr-1-his-1-(gly-1 or gly-2)-pur-1-pur-2. these markers may be analogous to hom, thra, hisa, glyc, and pura marker ... | 1986 | 3512533 |
| escherichia coli tyrosyl- and methionyl-trna synthetases display sequence similarity at the binding site for the 3'-end of trna. | covalent modification of escherichia coli tyrosyl-trna synthetase (tyrrs) by the 2',3'-dialdehyde derivative of trnatyr (trnaox) resulted in a time-dependent inactivation of both atp-ppi exchange and trna aminoacylation activities of the enzyme. in parallel with the inactivation, covalent incorporation of approximately 1 mol of [14c]trnatyrox/mol of the dimeric synthetase occurred. intact trnatyr protected the enzyme against inactivation by the trna dialdehyde. treatment of the tyrrs-[14c]trnaty ... | 1986 | 3513822 |
| internal thermodynamics of position 51 mutants and natural variants of tyrosyl-trna synthetase. | natural variation and evolution impose structural changes on an enzyme that can affect the energetics of catalysis. the energy profile of reaction could, in theory, be altered in three distinct ways: uniform binding changes, differential binding changes, and catalysis of elementary steps. residue threonine-51 of tyrosyl-trna synthetase from bacillus stearothermophilus is subject to natural variation, being replaced by alanine and proline in the enzymes from bacillus caldotenax and escherichia co ... | 1986 | 3518795 |
| a transcription terminator in the 5' non-coding region of the tyrosyl trna synthetase gene from bacillus stearothermophilus. | the 5' non-coding region of the tyrosyl-trna synthetase gene (tyrs) of bacillus stearothermophilus is 324 nucleotides long. it contains a premature terminator and a strong promoter: these were identified in vitro by rna run-off experiments and in escherichia coli by construction of specific mutants. the terminator consists of a stem and loop structure followed by the string of t residues characteristic of rho-independent termination. this is preceded by another stem and loop structure which may ... | 1986 | 3525162 |
| nucleotide sequence and characteristics of the gene for l-lactate dehydrogenase of thermus caldophilus gk24 and the deduced amino-acid sequence of the enzyme. | the gene for l-lactate dehydrogenase (ldh) (ec 1.1.1.27) of thermus caldophilus gk24 was cloned in escherichia coli using synthetic oligonucleotides as hybridization probes. the nucleotide sequence of the cloned dna was determined. the primary structure of the ldh was deduced from the nucleotide sequence. the deduced amino acid sequence agreed with the nh2-terminal and cooh-terminal sequences previously reported and the determined amino acid sequences of the peptides obtained from trypsin-digest ... | 1986 | 3533539 |
| mechanism of translational initiation in prokaryotes. evidence for a direct effect of if2 on the activity of the 30 s ribosomal subunit. | initiation factor if2 from either escherichia coli or bacillus stearothermophilus was found to possess the previously undetected property of stimulating the template-dependent ribosomal binding of aminoacyl-trnas with free alpha-nh2 groups. if1, which had no detectable activity alone, was found to stimulate the activity of e. coli if2 and, to a lesser extent, that of b. stearothermophilus if2. since in the absence of ribosomes not even a weak interaction between the two if2 molecules and the ami ... | 1986 | 3533628 |
| a new way of enhancing the thermostability of proteases. | thermostability of enzyme can be enhanced by single amino acid substitutions. recent advances in genetic engineering have made it possible to create novel proteins in a predictable manner where structural information for the protein is available. this 'protein engineering' has already been used to enhance enzyme thermostability, but it is usually not clear which amino acid substitutions should be made. we consider that the following approach should be helpful in engineering proteins with enhance ... | 1986 | 3540685 |
| characterization of the ultrastructure and the self-assembly of the surface layer of bacillus stearothermophilus strain nrs 2004/3a. | the ultrastructure of the crystalline surface layer (s-layer) of bacillus stearothermophilus strain nrs 2004/3a has been characterized by electron microscopy supplemented by optical and computer image analysis. the s-layer, composed of glycoprotein subunits, has oblique symmetry, and can be extracted by guanidine hydrochloride. upon dialysis, this extract produced both flat and cylindrical mono- and double-layer self-assembly products. optical diffraction analysis of negatively stained preparati ... | 1986 | 3453374 |
| crystal structure of a prokaryotic ribosomal protein. | the structure of ribosomal protein l30 from bacillus stearothermophilus has been solved to a resolution of 2.5 a. the molecule is somewhat elongated and contains two helices and a three-stranded, anti-parallel beta-pleated sheet. the protein fold, in which helices pack on the same side of the sheet, generates a simple helix-sheet, two-layered motif. it is possible to distinguish three hydrophobic patches on the molecular surface, and one end has six isolated arginine and lysine residues. it is p ... | 1986 | 3463963 |
| free energy of hydrolysis of tyrosyl adenylate and its binding to wild-type and engineered mutant tyrosyl-trna synthetases. | the equilibrium constant for the formation of tyrosyl adenylate and pyrophosphate from atp and tyrosine in solution has been measured by applying the haldane relationship to wild-type and three mutant tyrosyl-trna synthetases from bacillus stearothermophilus. the formation constant (=[tyr-amp] [ppi]/[atp] [tyr]) at ph 7.78, 25 degrees c, and 10 mm mgcl2 is (3.5 +/- 0.5) x 10(-7). this corresponds to a free energy of hydrolysis of tyrosyl adenylate at ph 7.0 and 25 degrees c of -16.7 kcal mol-1. ... | 1986 | 3466647 |
| the two alcohol dehydrogenases of zymomonas mobilis. purification by differential dye ligand chromatography, molecular characterisation and physiological roles. | the two alcohol dehydrogenases found in zymomonas mobilis have each been purified using dye-ligand chromatography and affinity elution with nucleotides. the isoenzyme with lower electrophoretic mobility (zadh-1) is a zinc enzyme with properties essentially similar to preparations described elsewhere. the faster isoenzyme (zadh-2) accounted for some 90% of the ethanol-oxidizing activity in freshly prepared extracts and corresponded to the iron-activated enzyme previously described. this enzyme wa ... | 1986 | 2935393 |
| drug residues in milk after intrauterine injection of oxytetracycline, lincomycin-spectinomycin, and povidone-iodine in cows with metritis. | a study was conducted to document the maximum retention times of antimicrobial residues in milk after their use in intrauterine treatment of metritis in lactating cows and to evaluate several risk factors hypothesized to influence the retention time of these drugs. oxytetracycline (3 g), lincomycin-spectinomycin (2 g of one-third lincomycin and two-thirds spectinomycin), or povidone-iodine (6 g) were given to cows with metritis by intrauterine route. the bacillus stearothermophilus var calidolac ... | 1986 | 2942064 |
| crystallographic structure of allosterically inhibited phosphofructokinase at 7 a resolution. | the structure of the allosterically inhibited form of phosphofructokinase from bacillus stearothermophilus has been determined by x-ray crystallography to 7 a resolution by molecular replacement using the known structure of the active state as a starting model. comparing the inhibited state with the active state, the tetramer is twisted about its long axis such that one pair of subunits in the tetramer rotates relative to the other pair by about 8 degrees around one of the molecular dyad axes. t ... | 1986 | 2949086 |
| temperature-sensitive binding of alpha-glucans by bacillus stearothermophilus. | bacillus stearothermophilus was found to bind strongly to starch and related alpha-glucans at 25 degrees c but not at 55 degrees c. the binding at the lower temperature could be assayed either by binding of fluorescein-labeled amylopectin to washed cell suspensions or through the reversible retention of bacteria by affinity chromatography in matrices containing immobilized starch. the bacteria exhibited amylopectin-dependent agglutination. the binding and agglutination were highest in bacteria g ... | 1986 | 3082861 |
| structure of a beta-galactosidase gene of bacillus stearothermophilus. | the nucleotide sequence of the bgab gene, which encodes the thermostable beta-galactosidase i of bacillus stearothermophilus, and its flanking region was determined. a 2,016-base-pair open reading frame observed was concluded to be for beta-galactosidase i (mr 78,051) from observations that the amino acid composition of the enzyme and the sequence of 14 amino acids from the amino-terminus of the enzyme coincided with those deduced from this open frame. a 107-base-pair haeiii-alui fragment just u ... | 1986 | 3086288 |
| cloning and nucleotide sequencing of genes for small, acid-soluble spore proteins of bacillus cereus, bacillus stearothermophilus, and "thermoactinomyces thalpophilus". | as found previously with other bacillus species, spores of b. stearothermophilus and "thermoactinomyces thalpophilus" contained significant levels of small, acid-soluble spore proteins (sasp) which were rapidly degraded during spore germination and which reacted with antibodies raised against b. megaterium sasp. genes coding for a b. stearothermophilus and a "t. thalpophilus" sasp as well as for two b. cereus sasp were cloned, their nucleotide sequences were determined, and the amino acid sequen ... | 1986 | 3087949 |
| construction of high, intermediate and low-copy-number promoter-probe plasmids for bacillus subtilis. | we constructed promoter-probe plasmids for bacillus subtilis based on the promoterless gene for penicillinase (penp) of bacillus licheniformis. a bacillus stearothermophilus dna fragment, which contained translational stop codons for all three reading frames and transcription terminators of the alpha-amylase gene, was inserted upstream from the shine-dalgarno sequence for the penp gene. a low-copy-number plasmid (about 10 copies per chromosome), ppf101, carried tetracycline-resistance gene, wher ... | 1986 | 3091455 |
| differential scanning calorimetry of bacteria. | thermograms obtained by differential scanning calorimetry of a range of bacteria of different heat resistances were compared. equations were derived to calculate the rate at which the numbers of viable organisms in a calorimeter decline as the temperature is raised at a constant rate. vegetative bacteria scanned at 10 degrees c min-1 showed multi-peaked thermograms with four major peaks (denoted m, n, p and q) occurring in the regions 68-73, 77-84, 89-99 and 105-110 degrees c respectively. excep ... | 1986 | 3093634 |
| the purification and characterization of glucokinase from the thermophile bacillus stearothermophilus. | homogeneous glucokinase (ec 2.7.1.2) from the thermophile bacillus stearothermophilus was isolated on the large scale by using four major steps: precipitation of extraneous material at ph 5.5, ion-exchange chromatography on deae-sepharose, pseudo-affinity chromatography on procion brown h-3r-sepharose 4b and gel filtration on ultrogel aca 34. the purified enzyme had a specific activity of about 330 units/mg of protein and was shown to exist as a dimer of subunit mr 33,000. kinetic parameters for ... | 1986 | 3099754 |
| isolation of a thermostable enzyme variant by cloning and selection in a thermophile. | we developed a method for rapidly generating thermostable enzyme variants. our strategy is to introduce the gene coding for a given enzyme from a mesophilic organism into a thermophile, bacillus stearothermophilus. variants that retain the enzymatic activity at the higher growth temperatures of the thermophile are then selected. this strategy was applied to kanamycin nucleotidyltransferase, which confers resistance to the antibiotic kanamycin. b. stearothermophilus carrying the wild-type enzyme ... | 1986 | 3003740 |
| construction of heterodimer tyrosyl-trna synthetase shows trnatyr interacts with both subunits. | the tyrosyl-trna synthetase (ec 6.1.1.1) from bacillus stearothermophilus is a dimer of two identical subunits. the dimer shows "half-of-the-sites" reactivity in that only one molecule of tyrosyladenylate is formed and one molecule of trnatyr binds per dimer. to identify whether the trnatyr binds to a single subunit in the dimer, or to both subunits, heterodimers were constructed by mixing two variant dimers together in 8 m urea. as the unfolded protein is electrophoresed into a native polyacryl ... | 1986 | 3006039 |
| structural genes encoding the thermophilic alpha-amylases of bacillus stearothermophilus and bacillus licheniformis. | the genes encoding the thermostable alpha-amylases of bacillus stearothermophilus and b. licheniformis were cloned in escherichia coli, and their dna sequences were determined. the coding and deduced polypeptide sequences are 59 and 62% homologous to each other, respectively. the b. stearothermophilus protein differs most significantly from that of b. licheniformis in that it possesses a 32-residue cooh-terminal tail. transformation of e. coli with vectors containing either gene resulted in the ... | 1986 | 3009417 |
| natural variation of tyrosyl-trna synthetase and comparison with engineered mutants. | we report the cloning and sequence analysis of the gene for the tyrosyl-trna synthetase from bacillus caldotenax and properties of the gene product. the amino acid sequence of the tyrosyl-trna synthetase was found to be 99% homologous with the corresponding enzyme from b. stearothermophilus, with only four amino acid differences. two of these natural variations were found to involve active site residues of the enzyme and correspond to mutations that have been engineered previously in vitro. one, ... | 1986 | 3011073 |
| thermostable alanine racemase from bacillus stearothermophilus: molecular cloning of the gene, enzyme purification, and characterization. | the alanine racemase (ec 5.1.1.1) gene of a thermophilic bacterium, bacillus stearothermophilus, was cloned and expressed in escherichia coli c600 with vector plasmid picr301, which was constructed from pbr322 and the l-alanine dehydrogenase gene derived from b. stearothermophilus. a coupled assay method with l-alanine dehydrogenase and tetrazolium salts was used to detect visually the alanine racemase activity in the clones. alanine racemase overproduced in a clone carrying the plasmid picr4, 1 ... | 1986 | 3015202 |
| rapid purification of glucokinase and glycerokinase from bacillus stearothermophilus by hydrophobic interaction chromatography. | 1986 | 3025236 | |
| molecular cloning and sequence of the bacillus stearothermophilus translational initiation factor if2 gene. | the structural gene for the bacillus stearothermophilus initiation factor if2 was localized to a 6 kb hindiii restriction fragment by cross-hybridization with the ssti-smai fragment of the escherichia coli infb gene. this fragment corresponds to the central region of the molecule containing the gtp-binding domain which is homologous in e. coli if2, ef-tu, ef-g and the human ras1 oncogene protein. after cloning into pacyc177, the hindiii fragment was further analysed by restriction mapping and cr ... | 1986 | 3025563 |
| cloning and complete nucleotide sequence of the bacillus stearothermophilus tryptophanyl trna synthetase gene. | the bacillus stearothermophilus nca 1503 tryptophanyl trna synthetase (wts; ec 6.1.1.2) gene has been cloned in escherichia coli and the amino acid (aa) sequence of the enzyme deduced unequivocally from the dna sequence of the cloned gene. the predicted aa sequence of the wts enzyme agrees with the previously determined aa sequence except that the dna sequence indicates a third arg residue at the c terminus of the enzyme over the two arg residues indicated by sequencing the protein itself. the t ... | 1986 | 3026925 |
| cloning, expression and complete nucleotide sequence of the bacillus stearothermophilus l-lactate dehydrogenase gene. | the structural gene for l-lactate dehydrogenase (ldh; ec 1.1.1.27) from bacillus stearothermophilus nca 1503 has been cloned in escherichia coli and its complete nucleotide sequence determined. the predicted amino acid (aa) sequence of the ldh enzyme agrees with the previously determined aa sequence except to three positions: aa 125 and 126, ser-glu, are inverted whilst his at position 130 has been replaced by ser in our sequence. the lct gene consists of an open reading frame (orf) commencing f ... | 1986 | 3026926 |
| effects of temperature on plasmid stability and penicillinase productivity of a transformant of bacillus stearothermophilus. | 1986 | 3488008 | |
| initial-rate studies of a thermophilic glucokinase from bacillus stearothermophilus. | the initial rates of phosphorylation of glucose catalysed by glucokinase from bacillus stearothermophilus were measured over a wide range of glucose, mgatp2-, mgadp- and glucose 6-phosphate concentrations. the results of the effects of the inhibitors on the initial rates suggest that the reaction mechanism is essentially the ordered bi bi, in which glucose adds to the enzyme before mgatp2- and glucose 6-phosphate is released from the enzyme after the dissociation of mgadp-, and also suggest that ... | 1987 | 3501715 |
| [extracellular thermostable alpha-amylase from bacillus stearothermophilus q8]. | the alpha-amylase of bacillus stearothermophilus q8, previously isolated in our laboratory, was purified by ammonium sulfate precipitation and cm-sephadex chromatography. the activity the of partial purified alpha-amylase was to be protected by bovine serum albumin ca2+ and mg2+. the enzyme showed 100% activity at ph 9.0; 98%, at ph 8.0 and 41%; at ph 10.0. it expressed optimal reaction temperature at 90 degrees c, 81% of the activities remained at 100 degrees c. after 15 min incubation at 100 d ... | 1987 | 3502403 |
| the use of a diffusion test for the detection of antibiotics in the tissues of slaughter stock. | the brilliant black reduction test kit (br test), which is widely used to detect antimicrobial residues in milk, was adapted to detect residues in the meat and tissues of slaughter stock. the adaptation consisted of placing kidney and muscle tissue samples into 2.5ml diffusion cups containing 0.4ml media plus bacillus stearothermophilus spores and brilliant black indicator. a preliminary trial undertaken to test the lower limits of sensitivity of the adapted br test to a number of the more commo ... | 1987 | 3508463 |
| a novel method for the synthesis of kyotorphin, tyr-arg, and 3h-tyr-arg, catalyzed by tyrosyl-trna synthetase from bacillus stearothermophilus. | a novel method of dipeptide synthesis is described that can be carried out in aqueous solution and does not require complicated protecting and deprotecting procedures. an analgesic neuropeptide named kyotorphin, h-tyr-arg-oh, was synthesized from unprotected tyrosine and arginine in a new enzymatic reaction catalyzed by immobilized tyrosyl-trna synthetase from bacillus stearothermophilus. the reaction could be a useful tool in the syntheses of radioisotope-labeled oligopeptides to be used in rec ... | 1987 | 3509140 |
| investigation of transition-state stabilization by residues histidine-45 and threonine-40 in the tyrosyl-trna synthetase. | we have analyzed various mutations involving residues thr-40 and his-45 in the tyrosyl-trna synthetase of bacillus stearothermophilus. the utilization of binding energy in catalysis of tyrosyl adenylate formation from tyrosine and atp was determined from the free energy profiles for the mutant enzymes. our results confirm that the side chains of thr-40 and his-45 provide a binding site for the pyrophosphoryl portion of the transition state of this reaction and for pyrophosphate in the reverse re ... | 1987 | 3126804 |
| the primary structure of the ribosomal a-protein (l12) from the halophilic eubacterium haloanaerobium praevalens. | the ribosomal a-protein, equivalent to the ribosomal protein l12 from escherichia coli, has been sequenced from the anaerobic halophilic eubacterium haloanaerobium praevalens (dsm 2228). the protein contains 122 amino acids, has a composition of asp6, asn2, thr2, ser6, glu22, pro2, gly13, ala19, val12, met4, ile5, leu11, phe3, lys14, arg1 and has a molecular weight of 12,691. the hydrophilicity profile was determined for this protein. a phylogenetic or cluster tree was calculated from computer a ... | 1987 | 3126821 |
| sequence of the bacillus caldotenax and bacillus stearothermophilus lctb genes. | 1987 | 3029705 | |
| cloning and nucleotide sequencing of genes for a second type of small, acid-soluble spore proteins of bacillus cereus, bacillus stearothermophilus, and "thermoactinomyces thalpophilus". | the nucleotide sequences of the single genes coding for the b-type small, acid-soluble spore proteins (sasp) of bacillus cereus, b. stearothermophilus, and "thermoactinomyces thalpophilus" were determined, and the amino acid sequences of all b-type sasp were compared. while this type of sasp showed significant sequence conservation around the two spore protease cleavage sites, alignment of these sequences required the introduction of gaps, and even then only 19 of the residues were conserved exa ... | 1987 | 3036769 |
| 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 |
| 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 |
| 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 |
| [dependence of microbiologic test results of formaldehyde gas sterilization methods on the nature of the test material]. | the efficiency of a formaldehyde gas sterilization procedure was evaluated with the aid of test pieces consisting of various materials. both rigid and flexible tubes served as test pieces. the tubes were 75 cm long with an inner diameter of 1 mm and were sealed at one end. the bioindicators were placed inside the tubes close to the sealed end. dried spores of bacillus stearothermophilus adhering to linen threads served as test organisms. the test results varied according to the material of the t ... | 1987 | 3113100 |
| a strong carboxylate-arginine interaction is important in substrate orientation and recognition in lactate dehydrogenase. | using site-directed mutagenesis, arginine-171 at the substrate-binding site of bacillus stearothermophilus, lactate dehydrogenase has been replaced by lysine. in the closely homologous eukaryotic lactate dehydrogenase, this residue binds the carboxylate group of the substrate by forming a planar bifurcated bond. the mutation diminishes the binding energy of pyruvate, alpha-ketobutyrate and alpha-ketovalerate (measured by kcat/km) by the same amount (about 6 kcal/mol). for each additional methyle ... | 1987 | 3113484 |
| pathogenic capacity of proteases from serratia marcescens and pseudomonas aeruginosa and their suppression by chicken egg white ovomacroglobulin. | the pathogenicities of three proteases from serratia marcescens, two proteases from pseudomonas aeruginosa, and one thermolysin from bacillus stearothermophilus were examined. all proteases tested caused acute liquefactive necrosis of the cornea and descemetocele formation in guinea pig eyes after intrastromal injection, with the exception of the 60-kilodalton protease from s. marcescens, which produced only an opaque lesion. when injected into guinea pig skin, the protease also enhanced vascula ... | 1987 | 3115900 |
| synthesis and secretion of a heat-stable carboxymethylcellulose from clostridium thermocellum in bacillus subtilis and bacillus stearothermophilus. | the cellulase gene cela of clostridium thermocellum coding for the thermostable endoglucanase a was transferred from escherichia coli to bacillus subtilis 168 and b. stearothermophilus cu21 using plasmids derived from the bacillus vector pub110. when the structural part of the gene was joined to a pub110 promoter the recombinant plasmids (pse102, pse105) were stably maintained and expressed carboxymethylcellulose (cmcase) activity. in b. stearothermophilus cu21 (pse105) the clostridial cmcase wa ... | 1987 | 3118140 |
| antimicrobial activity of lysozyme against bacteria involved in food spoilage and food-borne disease. | egg white lysozyme was demonstrated to have antibacterial activity against organisms of concern in food safety, including listeria monocytogenes and certain strains of clostridium botulinum. we also found that the food spoilage thermophile clostridium thermosaccharolyticum was highly susceptible to lysozyme and confirmed that the spoilage organisms bacillus stearothermophilus and clostridium tyrobutyricum were also extremely sensitive. several gram-positive and gram-negative pathogens isolated f ... | 1987 | 3118808 |
| three-dimensional crystals of ribosomes and their subunits from eu- and archaebacteria. | ordered three-dimensional crystals of 70s ribosomes as well as of 30s and 50s ribosomal subunits from various bacteria (e. coli, bacillus stearothermophilus, thermus thermophilus and halobacterium marismortui) have been grown by vapour diffusion in hanging drops using mono- and polyalcohols. a new compact crystal form of 50s subunits has been obtained, and it is suitable for crystallographic studies at medium resolution. in addition, from one crystal form large crystals could be grown in x-ray c ... | 1987 | 3124853 |
| 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 |
| structure-activity relationships in engineered proteins: characterization of disruptive deletions in the alpha-ammonium group binding site of tyrosyl-trna synthetase. | residues asp-78 and gln-173 of the tyrosyl-trna synthetase of bacillus stearothermophilus form part of the binding site for tyrosine by making hydrogen bonds with the alpha-ammonium group. asp-38 is close enough to the group to make an important electrostatic contribution. unlike other residues in the active site that have been studied by site-directed mutagenesis, asp-38, asp-78, and gln-173 are part of hydrogen-bonded networks. each of these residues has been mutated to an alanine, and the res ... | 1987 | 3480006 |
| isolation and structure determination of a diacetamidodideoxyuronic acid-containing glycan chain from the s-layer glycoprotein of bacillus stearothermophilus nrs 2004/3a. | a glycan isolated from the surface-layer glycoprotein of bacillus stearothermophilus strain nrs 2004/3a was shown by 1h- and 13c-n.m.r. spectroscopy to have the tetrasaccharide repeating-unit ----4)-beta-manpa2,3(nac)2-(1----3)-alpha-glcpnac-(1----4)-beta- manpa2,3(nac)-(1----6)-alpha-glcp(1----. | 1987 | 3427581 |
| the active site of manganese-containing superoxide dismutase from bacillus stearothermophilus studied by 1h and 19f magnetic relaxation. | the dependence of the magnetic relaxation rates of 1h and 19f- on temperature, frequency, ph and n-3 concentration, were measured in solutions of manganese-containing superoxide dismutase of bacillus stearothermophilus, and were compared to activity measurements, in order to obtain some information on the structure and dynamics at mn(iii) present in the active site of the enzyme. the experimental data lead us to hypothesize the presence of two binding sites in the coordination sphere of the enzy ... | 1987 | 3428245 |
| relation of the heat resistance of bacterial spores to chemical composition and structure. ii. relation to cortex and structure. | the relation between the amount of cortex, measured as total hexosamine, as diaminopimelic acid and as muramic lactam, and the heat resistance of spores of five different strains of bacillus stearothermophilus was studied. electron micrographs of thin sections of the spores were made to relate the structure of the spores to chemical and thermal characteristics. it was found that the amount of the cortex was significantly related to heat resistance of the spores. strains with more electron-dense ... | 1987 | 3429356 |
| the growth of ordered two-dimensional sheets of ribosomal particles from salt-alcohol mixtures. | a procedure for the in vitro growth of well-ordered two-dimensional sheets from ribosomal particles using salts and salt-alcohol mixtures has been developed. employing this procedure, ordered two-dimensional sheets of the wild type as well as of mutated 50 s ribosomal subunits from bacillus stearothermophilus can readily be obtained. these sheets, stained with uranyl acetate or gold-thioglucose, are suitable for three-dimensional image reconstruction. they consist of relatively small unit cells ... | 1987 | 3434787 |
| reconstitution and crystallisation experiments with isolated split proteins from bacillus stearothermophilus ribosomes. | six proteins (b-l1, b-l6, b-l10, b-l11, b-l12 and b-l16) were removed from 50s ribosomal subunits of bacillus stearothermophilus by treatment with ethanol and ammonium chloride. the proteins were isolated in a pure form, and one of them (b-l6) was crystallized. five of the six proteins (in various combinations) were added back to the core particles, resulting in 50s subunits lacking one protein. the biological activities of these ribosomal particles as determined in the poly(u)-system varied ove ... | 1987 | 3435552 |
| order of binding of substrate to valyl-trna synthetase from bacillus stearothermophilus in amino acid activation reaction. | amino acid activation reaction with valyl-trna synthetase (ec 6.1.1.9) from bacillus stearothermophilus was studied kinetically by measuring atp-ppi exchange to find the order of the binding of substrate to the enzyme. the effects of the concentration of the substrates (l-valine and atp) and two dead-end inhibitors (l-valinol and adenosine) on the reaction rate were analyzed. the results indicate that l-valine and atp are bound to the enzyme in a random sequence. this conclusion is consistent wi ... | 1987 | 3453086 |
| 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 |
| protein processing to form extracellular thermostable alpha-amylases from a gene fused in a bacillus subtilis secretion vector. | a thermostable alpha-amylase gene (amyt631) from bacillus stearothermophilus a631 was cloned into pbr322 and recloned into pub110: the resulting plasmid was designated ptub607. to investigate the processing from preproenzyme to mature enzyme, amyt631 from ptub607, after digestion with bal31, was introduced into the b. subtilis alpha-amylase secretion vector ptub285. three chimaeric plasmids, ptub613, ptub616, and ptub617, were isolated. the fused alpha-amylases expressed from the three plasmids ... | 1987 | 3128640 |