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molecular cloning and nucleotide sequence of the glycogen branching enzyme gene (glgb) from bacillus stearothermophilus and expression in escherichia coli and bacillus subtilis.the structural gene for the bacillus stearothermophilus glycogen branching enzyme (glgb) was cloned in escherichia coli. nucleotide sequence analysis revealed a 1917 nucleotide open reading frame (orf) encoding a protein with an mr of 74787 showing extensive similarity to other bacterial branching enzymes, but with a shorter n-terminal region. a second orf of 951 nucleotides encoding a 36971 da protein started upstream of the glgb gene. the n-terminus of the orf2 gene product had similarity to t ...19911745226
determination of peptide regions on the surface of the eubacterial and archaebacterial ribosome by limited proteolytic digestion.limited proteolysis was used in combination with two-dimensional gel electrophoresis, blotting, and amino acid sequence analysis to investigate the surface of intact ribosomal subunits at the peptide and amino acid level. surface sites of 14 ribosomal proteins from escherichia coli 50s subunits were determined using proteases with different specificities. to assess the evolutionary conservation of ribosomal topography among eubacteria, large subunits from bacillus stearothermophilus were also su ...19911751495
sequence and expression of the gene encoding 3-phosphoglycerate kinase from bacillus stearothermophilus.the structural gene (pgk) encoding 3-phosphoglycerate (pgk) from bacillus stearothermophilus nca1503, has been cloned in escherichia coli and its complete nucleotide sequence determined. the gene consists of an open reading frame corresponding to a protein of 394 amino acids (aa) (calculated mr 42,703) and, in common with other prokaryotic pgk genes, is preceded by the structural gene encoding glyceraldehyde-3-phosphate dehydrogenase (gapdh). constructs containing the b. stearothermophilus pgk g ...19911756980
bacillus stearothermophilus disk assay for determining ampicillin residues in fish muscle.the bacillus stearothermophilus disk assay for penicillin in milk (aoac official method) was adapted for the determination of ampicillin in fish muscle. the method was evaluated in 2 species of cultured fish: channel catfish and striped bass. recoveries of ampicillin ranged from 99 to 104% when muscle specimens from both species were spiked at concentrations of 0.025-1.00 micrograms/g. the lower limit of determination (lod) was 0.025 micrograms/g. the assay was applied to monitor the elimination ...19911757413
thermostable alanine racemase of bacillus stearothermophilus: subunit dissociation and unfolding.the guanidine hydrochloride-induced subunit dissociation and unfolding of thermostable alanine racemase from bacillus stearothermophilus have been studied by circular dichroism, fluorescence and absorption spectroscopies, and gel filtration. the overall process was found to be reversible: more than 75% of the original activity was recovered upon reduction of the denaturant concentration. in the range of 0.6 to 1.5 m guanidine hydrochloride, the dimeric enzyme was dissociated into a monomeric for ...19911761523
primary structures of ribosomal proteins from the archaebacterium halobacterium marismortui and the eubacterium bacillus stearothermophilus.approximately 40 ribosomal proteins from each halobacterium marismortui and bacillus stearothermophilus have been sequenced either by direct protein sequence analysis or by dna sequence analysis of the appropriate genes. the comparison of the amino acid sequences from the archaebacterium h marismortui with the available ribosomal proteins from the eubacterial and eukaryotic kingdoms revealed four different groups of proteins: 24 proteins are related to both eubacterial as well as eukaryotic prot ...19911764513
the amino-acid sequences of the bacillus stearothermophilus ribosomal proteins s17 and s21 and their comparison to homologous proteins of other ribosomes.ribosomal proteins s17 and s21 from the moderate thermophile bacillus stearothermophilus were purified by one-step high-performance liquid chromatography from the 30s-subunit protein mixture employing a semi-preparative reversed-phase c4 column. the complete amino-acid sequences of these proteins were determined by a combination of n-terminal sequencing in picomole quantities of the protein and of appropriate peptide fragments. proteins s17 and s21 consist of 86 and 55 amino-acid residues, corre ...19911772592
sequencing with the large fragment of dna polymerase i from bacillus stearothermophilus.the large fragment of dna polymerase i, isolated from bacillus stearothermophilus, was used for dideoxy sequencing. this heat-stable enzyme permits performing sequencing reactions at high temperature to melt secondary structure and results in uniform band intensities and low background on the autoradiogram. the enzyme can be used in the standard sanger one-step protocol or in a two-step protocol which separates the labeling reaction from the elongation-termination reaction. the enzyme can be use ...19911773056
cloning and expression of an amylase gene from bacillus stearothermophilus.in the industrial process of liquefying starch to make glucose or high fructose syrups, it is crucial that the amylase used is stable and active at about 105 degrees c at ph 6.5 or preferentially at a lower ph. the amylase from bacillus licheniformis is well suited for this purpose but it is possible that other amylases might perform even better. therefore, we cloned and characterized amys encoding a heat-stable alpha-amylase from bacillus stearothermophilus. using a newly developed method for c ...19911784818
development and evaluation of a method to determine indicator microorganisms in air emissions and residue from medical waste incinerators.to allow testing of microbial destruction in medical waste incinerators, methods were developed to determine indicator microorganisms (bacillus stearothermophilus spores) in incinerator air emissions and residue. the emission trapping train consisted of a water cooled glass probe and impingers containing a neutral phosphate buffer. in field tests, spores were injected directly into the probe, and results showed that approximately 60 percent of the spores were recovered. spores were analyzed with ...19911789955
large fragment of dna polymerase i from bacillus stearothermophilus (bst polymerase) is stable at ambient temperature. 19911793578
the structural domains in the e2 component of the pyruvate dehydrogenase multienzyme complex from bacillus stearothermophilus. 19911794583
[determination of antibiotic chemicals using microbiological tests: evaluation of the limits of sensitivity].sensitivity of bacillus subtilis bga and bacillus stearothermophilus var. calidolactis disc assays to 53 chemio-antibiotics was tested. test-microorganisms were sown in two different mediums: pm indicator agar, difco u.s.a., and standard ii nutrient agar, merck germany, modified according to nouws. the mediums were used with or without addition of trimethoprim (at a concentration of 0.12 or 0.024 mcg/ml of medium for b. subtilis and for b. stearothermophilus respectively). b. stearothermophilus ...19911804237
isolation of a new antitumor substance from bacillus stearothermophilus.a new antitumor substance, bs-1, was isolated from the autolysate and culture filtrate of bacillus stearothermophilus uk563 by ethylacetate extraction and hplc. bs-1 inhibited the proliferation of mouse macrophage-like cells, p388-d1 (ic50: 4 micrograms/ml) and mouse mastocytoma, p-815 (ic50: 0.6 microgram/ml), but not that of balb/c 3t3.19911804563
sterilization beneath rings on dental instruments.this study determined the effectiveness of standard methods of instrument sterilization beneath instrument rings. sets of three types of dental instruments were contaminated with known amounts of bacterial spores (bacillus stearothermophilus or bacillus subtilis). instrument rings were placed over the contamination and the instruments processed through standard cycles in a steam autoclave, an unsaturated chemical vapor sterilizer, a standard dry heat sterilizer, an ethylene oxide gas sterilizer ...19911814351
stabilization of the neutral protease of bacillus stearothermophilus by removal of a buried water molecule.using site-directed mutagenesis, ala166 in the neutral protease of bacillus stearothermophilus was changed into ser. model building and molecular dynamics simulations of the mutant enzyme indicated that the ser hydroxyl group fits well in a cavity which contains a water molecule in the wild-type enzyme. the ala166----ser mutation was expected to exert a stabilizing effect because of the gain in entropy resulting from the release of water molecule from the folded protein to the solvent. in additi ...19911817257
purification and properties of the phosphofructokinase from lactobacillus bulgaricus. a non-allosteric analog of the enzyme from escherichia coli.phosphofructokinase, the enzyme which catalyzes the conversion of fructose 6-phosphate into fructose 1,6-bisphosphate in lactobacillus bulgaricus (lactobacillus delbrueckii, subspecies bulgaricus) has been purified to homogeneity and some of its structural and functional properties have been studied. the enzyme is a tetramer composed of four 35-kda subunits. its n-terminal sequence determined on 38 residues is homologous to those of the major allosteric enzymes from escherichia coli and bacillus ...19911828763
characterization of bacillus stearothermophilus cyclodextrin glucanotransferase in ascorbic acid 2-o-alpha-glucoside formation.in this study, we characterized cyclodextrin glucanotransferase (cgtase) from bacillus stearothermophilus in l-ascorbic acid-2-o-alpha-d-glucoside (aa-2g) formation and compared its enzymological properties with those of rat intestinal and rice seed alpha-glucosidases which had the ability to form aa-2g. cgtase formed aa-2g efficiently using alpha-cyclodextrin (alpha-cd) as a substrate and ascorbic acid (aa) as an acceptor. several aa-2-oligoglucosides were also formed in this reaction mixture, ...19911829640
bstbsi, a restriction endonuclease from bacillus stearothermophilus bs which recognizes 5'gtatac3'. 19911831259
isolation of bacillus stearothermophilus constituent that suppresses mouse mixed leukocyte reaction in vitro.a novel immunosuppressant, fr.5-b, has been isolated from the debris of bacillus stearothermophilus uk563 autolysate. fr.5-b suppressed mouse mixed leukocyte reaction at doses ranging from 10(-4) to 1 microgram/ml, but not proliferation stimulated by concanavalin a and lipopolysaccharide.19911839973
cloning and analysis of the beta-galactosidase-encoding gene from clostridium thermosulfurogenes em1.clostridium thermosulfurogenes em1 produced a thermostable (up to 70 degrees c) beta-galactosidase (beta gal) with a ph optimum of 7 during growth on lactose. the gene (lacz) encoding this enzyme was cloned and expressed in escherichia coli using puc18 as a vector. the nucleotide sequence of a 2.7-kb psti fragment carrying the lacz gene was determined. the open reading frame for lacz, which encoded a protein of 716 amino acids with a calculated mr of 83,728, was confirmed by the identity of its ...19911840542
methionyl-trna synthetase from bacillus stearothermophilus: structural and functional identities with the escherichia coli enzyme.the mets gene encoding homodimeric methionyl-trna synthetase from bacillus stearothermophilus has been cloned and a 2880 base pair sequence solved. comparison of the deduced enzyme protomer sequence (mr 74,355) with that of the e. coli methionyl-trna synthetase protomer (mr 76,124) revealed a relatively low level (32%) of identities, although both enzymes have very similar biochemical properties (kalogerakos, t., dessen, p., fayat, g. and blanquet, s. (1980) biochemistry 19, 3712-3723). however, ...19911852609
cloning and characterization of a glutamine transport operon of bacillus stearothermophilus nub36: effect of temperature on regulation of transcription.we cloned and sequenced a fragment of the bacillus stearothermophilus nub36 chromosome that contains two open reading frames (orfs) whose products were detected only in cells of cultures grown in complex medium at high temperature. the nucleotide sequence of the two orfs exhibited significant identity to the sequence of the glnq and glnh loci of the glutamine transport system in enteric bacteria. in addition, growth response to glutamine, sensitivity to the toxic glutamine analog gamma-l-glutamy ...19911856180
temperature-induced protein synthesis in bacillus stearothermophilus nub36.cultures of bacillus stearothermophilus subjected to a temperature shift-up or shift-down of 15 degrees c within the normal temperature range of growth (45 to 65 degrees c) enter a transient adaptation period before exponential growth at the new temperature. the de novo synthesis of some proteins coincides with the adaptation period.19911856181
cysteinyl-trna synthetase is a direct descendant of the first aminoacyl-trna synthetase.the gene encoding the cysteinyl-trna synthetase of e. coli was cloned from an e. coli genomic library made in lambda 2761, a lambda vector which can integrate and which carries a chloramphenicol resistance gene. a thermosensitive cyss mutant of e. coli was lysogenised and chloramphenicol-resistant colonies able to grow at 42 degrees c were selected to isolate phages containing the wild-type cyss gene. the sequence of the gene was determined. it codes for a 461 amino-acid protein and includes the ...19911864365
cloning, overexpression, purification and crystallisation of ribosomal protein l9 from bacillus stearothermophilus.the cloning, sequencing and overexpression of the gene coding for bacillus stearothermophilus ribosomal protein l9 is described. the sequence corresponds directly to that presented for the protein itself by classical methods, differing at only a few amino acid positions. the purification and crystallisation of the corresponding l9 protein is presented. the crystals are isomorphous to those described for l9 obtained by conventional methods.19911864369
cloning the bstvi restriction-modification system in escherichia coli.a standard dna modification methyltransferase (mtase) selection protocol was followed to clone the bstvi restriction and modification system from bacillus stearothermophilus in escherichia coli. both genes were contained in a 4.4-kb ecori fragment from b. stearothermophilus v chromosomal dna. the heterologous expression of these genes did not depend on their orientation in the vector, suggesting that the genes are expressed in e. coli under the control of promoters located on the cloned fragment ...19911864512
the primary structure of rat ribosomal protein s18.the amino acid sequence of the rat 40s ribosomal subunit protein s18 was deduced from the sequence of nucleotides in a recombinant cdna. s18 has 152 amino acids and has a molecular weight of 17,707. hybridization of the cdna to digests of nuclear dna suggests that there are 10-13 copies of the s18 gene. the mrna for the protein is about 600 nucleotides in length. rat s18 is identical to mouse s18 (also referred to as ke3) and is related to escherichia coli s13 and to other s13-like ribosomal pro ...19911872840
characterization and comparative sequence analysis of replication origins from three large bacillus thuringiensis plasmids.the replication origins of three large bacillus thuringiensis plasmids, derived from b. thuringiensis hd263 subsp. kurstaki, have been cloned in escherichia coli and sequenced. the replication origins, designated ori 43, ori 44, and ori 60, were isolated from plasmids of 43, 44, and 60 mda, respectively. each cloned replication origin exhibits incompatibility with the resident b. thuringiensis plasmid from which it was derived. recombinant plasmids containing the three replication origins varied ...19911885511
molecular dissection of translation initiation factor if2. evidence for two structural and functional domains.by means of limited proteolysis of bacillus stearothermophilus initiation factor if2 and genetic manipulation of its structural gene, infb, we have been able to produce (or hyperproduce) and purify two polypeptide fragments corresponding to two structurally and functionally separate domains of the protein. the first is the g-domain (approximately 41 kda), which makes up the central part of the molecule and contains the conserved structural elements found in all gtp/gdp-binding sites of g-protein ...19911885570
competitive inhibition of liver glucokinase by its regulatory protein.the regulatory protein of rat liver glucokinase (hexokinase iv or d) behaved as a fully competitive inhibitor of this enzyme when glucose was the variable substrate, i.e. it increased the half-saturating concentration of glucose as a linear function of its concentration without affecting v (velocity at infinite concentration of substrate). the inhibition by the regulatory protein and that by palmitoyl-coa were synergistic with that by n-acetyl-glucosamine, indicating that the two former inhibito ...19911889417
physical studies on membrane lipids of bacillus stearothermophilus temperature and calcium effects.bacillus stearothermophilus was grown at the optimal temperature range (center, 65 degrees c), below it (48 and 55 degrees c), and above it (68 degrees c), in a complex medium with or without 2.5 mm ca2+. the ca(2+)-supplement improves growth at sub- and supraoptimal temperatures and extends it to higher temperatures (jurado et al. (1987) j. gen. microbiol. 133, 507-513). the phospholipid composition of cultures obtained in the different growth conditions was studied. phosphatidylethanolamine wa ...19911898060
evidence that gene g7a in the human major histocompatibility complex encodes valyl-trna synthetase.at least 36 genes have now been located in a 680 kb segment of dna between the class i and class ii multigene families within the class iii region of the human major histocompatibility complex on chromosome 6p21.3. the complete nucleotide sequence of the 4.3 kb mrna of one of these genes, g7a (or bat6), has been determined from cdna and genomic clones. the single-copy g7a gene encodes a 1265-amino-acid protein of molecular mass 140,457 da. comparison of the derived amino acid sequence of the g7a ...19911898367
molecular cloning, sequencing, and identification of a metalloprotease gene from listeria monocytogenes that is species specific and physically linked to the listeriolysin gene.the entire nucleotide sequence of an open reading frame located immediately downstream of the listeriolysin gene from a virulent listeria monocytogenes serotype 1/2a strain was determined. the product of the open reading frame was 510 amino acids with a predicted molecular weight of 57,400. the deduced amino acid sequence of this open reading frame is highly similar to that of a family of secreted metalloproteases produced by various members of the genus bacillus, of which thermolysin is the pro ...19911898903
cloning of a chromosomal alpha-amylase gene from bacillus stearothermophilus.we have cloned and sequenced a gene for a heat-stable alpha-amylase from a natural isolate of bacillus stearothermophilus. previously, it had been shown that b. stearothermophilus amylase genes may be harboured on indigenous plasmids. we have found that our isolate harbours the amylase gene only on the chromosome and not on its indigenous plasmid.19911903751
a highly thermostable neutral protease from bacillus caldolyticus: cloning and expression of the gene in bacillus subtilis and characterization of the gene product.by using a gene library of bacillus caldolyticus constructed in phage lambda embl12 and selecting for proteolytically active phages on plates supplemented with 0.8% skim milk, chromosomal b. caldolyticus dna fragments that specified proteolytic activity were obtained. subcloning of one of these fragments in a protease-deficient bacillus subtilis strain resulted in protease proficiency of the host. the nucleotide sequence of a 2-kb hinfi-mlui fragment contained an open reading frame (orf) that sp ...19911905714
thermostable alanine racemase of bacillus stearothermophilus. construction and expression of active fragmentary enzyme.limited proteolysis studies on alanine racemase suggested that the enzyme subunit is composed of two domains (galakatos, n. g., and walsh, c. t. (1987) biochemistry 26, 8475-8480). we have constructed a mutant gene that tandemly encodes the two polypeptides of the bacillus stearothermophilus enzyme subunit cleaved at the position corresponding to the predicted hinge region. the mutant gene product purified was shown to be composed of two sets of the two polypeptide fragments and was immunologica ...19911906880
duck liver malic enzyme: sequence of a tryptic peptide containing the cysteine residue labeled by the substrate analog bromopyruvate.malic enzyme of duck liver is alkylated by bromopyruvate with half-of-the-sites stoichiometry, and with accompanying loss of oxidative decarboxylase and enhancement of pyruvate reductase activities as was previously shown for the pigeon enzyme (hsu, r.y. (1982) mol. cell. biochem. 43, 3-26). in the present work, the alkylated enzyme is shown to bind nadph, but not l-malate in the presence of mncl2, indicating impairment of the enzyme site for the substrate and/or divalent metal. the enzyme was d ...19911911848
sequence-specific 1h-nmr assignments and secondary structure of the lipoyl domain of the bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex.the lipoyl domain (residues 1-85) of the lipoate-acetyltransferase polypeptide chain of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been subjected to detailed structural analysis by means of two-dimensional (2d) 1h-nmr spectroscopy at 400 mhz. sequence-specific proton resonance assignments were made, but at this field strength not all of the side-chain protons could be assigned, especially from complex spin systems like those of leucine, proline and lysine r ...19911915365
analysis of the active center of bacillus stearothermophilus neopullulanase.the active center of the neopullulanase from bacillus stearothermophilus was analyzed by means of site-directed mutagenesis. the amino acid residues located in the active center of the neopullulanase were tentatively identified according to a molecular model of taka-amylase a and homology analysis of the amino acid sequences of neopullulanse, taka-amylase a, and other amylolytic enzymes. when amino acid residues glu and asp, corresponding to the putative catalytic sites, were replaced by the opp ...19911917847
growth kinetics of bacillus stearothermophilus br219.bacillus stearothermophilus br219, a phenol-resistant thermophile, can convert phenol to the specialty chemical catechol. the growth kinetics of this organism were studied in batch, continuous, and immobilized-cell culture. batch growth was insensitive to ph between 6.0 and 8.0, but little growth occurred at 5.5. in continuous culture on a dilute medium supplemented with 10 mm phenol, several steady states were achieved between dilution rates of 0.25 and 1.3 h-1. phenol degradation was found to ...19911929366
comparison of the structure of archaebacterial ribosomal proteins equivalent to proteins l11 and l1 from escherichia coli ribosomes.the sequences of two ribosomal proteins from two widely divergent species of archaebacteria, halobacterium cutirubrum and sulfolobus solfataricus, have been deduced from the structure of their respective genes. these two proteins were found to be equivalent to the l11 and l1 ribosomal proteins of the eubacterium escherichia coli. sequence comparison revealed that the archaebacterial l11e (equivalent to e. coli l11) proteins are longer than the eubacterial protein due to a c-terminal extension of ...19911946333
influence of an s-layer on surface properties of bacillus stearothermophilus.various aspects of surface properties of the s-layer-carrying bacillus stearothermophilus pv72 and of an s-layer-deficient mutant (strain pv72/t5) have been tested by adsorption assays on solid surfaces, electrostatic interaction chromatography and hydrophobic interaction chromatography. the adsorption assays have shown that cell adhesion of the s-layer-carrying strain was less influenced by environmental changes than it was with the s-layer-deficient mutant. electrostatic interaction chromatogr ...19911953302
effect of simultaneous application of heat and pressure on the survival of bacterial spores.the effect of simultaneous application of moderate hydrostatic pressure (10-300 atm) and heat on the survival of the bacillus stearothermophilus spores in a flow-through system was investigated. a high heterogeneity of the sensitization of spores to heat by pressure was found. a higher degree of reduction of heat resistance was observed at the low than at the high temperatures tested. the simultaneous application of moderate pressure and heat can not be applied for the preservation of liquid foo ...19911955421
single-stranded dna plasmid, vector construction and cloning of bacillus stearothermophilus alpha-amylase in lactobacillus.vector plasmids were constructed by ligating chloramphenicol and erythromycin resistance genes to taqi-digested dna of a cryptic plasmid from lactobacillus plantarum. the minimal region of lactobacillus plasmid dna that was required for dna replication was defined and a single-stranded dna intermediate replication system was observed. homologies with other origins of replication of plasmids from gram-positive bacteria, replicating via rolling circle mechanism, were found. it was shown that the c ...19911961976
cloning, sequencing, and overexpression of genes for ribosomal proteins from bacillus stearothermophilus.although a low resolution model for the arrangement of the proteins of the small and large ribosomal subunits is known, a detailed mechanistic understanding of the function of the ribosome awaits a high resolution structure of its components. while crystals have been obtained of several ribosomal proteins from bacillus stearothermophilus, determination of atomic resolution structures of these proteins is impeded by the difficulty of obtaining large amounts of native proteins for crystallographic ...19911985969
detection and characterization of intermediates in the folding of large proteins by the use of genetically inserted tryptophan probes.l-lactate dehydrogenase from bacillus stearothermophilus was rebuilt by using site-directed mutagenesis to produce an enzymically active, tryptophan-less enzyme by replacing all the wild-type tryptophans (80, 150, and 203) by tyrosines. nine single tryptophan-containing active enzymes were constructed from this enzyme by genetically replacing one of the tyrosines 36, 85, 147, 190, 203, 237, 248, 279, or 285 by tryptophan. the equilibrium and the time-resolved tryptophan fluorescence intensity an ...19911989674
the identification of a structurally important cysteine residue in the glycerol dehydrogenase from bacillus stearothermophilus.evidence is presented to demonstrate that the zn2+ metallo-enzyme glycerol dehydrogenase from the thermophile bacillus stearothermophilus has one cysteine residue per subunit which is only available for reaction with thiol reagents in the metal-depleted form of the enzyme. modification of the metal-depleted enzyme by methyl methanethiosulphonate prevents the reactivation of the enzyme by zn2+ ions and induces dissociation of the oligomer into subunits. the rate of reaction of the cysteine residu ...19912009285
fluidity of bacterial membrane lipids monitored by intramolecular excimerization of 1.3-di(2-pyrenyl)propane.intramolecular excimer formation of 1,3-di(2-pyrenyl)propane was used to study the fluidity of liposomes prepared from membrane polar lipids of bacillus stearothermophilus. on the basis of spectral data, local polarity and polarizability parameters were established suggesting that the probe molecules are located well inside the membranes, but displaced towards the polar head groups of the phospholipid molecules. the excimerization rate is very sensitive to lipid phase transitions and pretransiti ...19912018528
promoter sequence analysis in bacillus and escherichia: construction of strong promoters in e. coli.many derivatives of the nprm promoter of bacillus stearothermophilus and the strong early promoter, a3, of coliphage t3 were designed and chemically synthesized. these promoters consisted of some or all of the at box, consensus sequence, tac promoter sequence, spacer, and lac operator. the promoter activities were assessed by their ability to express the cat gene in escherichia coli. one of the derivatives of the a3 promoter, which contained the lac operator, was much stronger (about 3.5 times) ...19912022318
improving the thermostability of the neutral protease of bacillus stearothermophilus by replacing a buried asparagine by leucine.amino acids buried in the hydrophobic interior of a protein with polar side chain atoms, which are not involved in hydrogen bonding or electrostatic interactions, have an adverse effect on protein stability. replacing such residues by hydrophobic ones may render a protein more stable. asparagine 241, which is buried in the neutral protease of bacillus stearothermophilus, was replaced by leucine by site-directed mutagenesis. this mutation increased the stability of the protein by 0.7 +/- 0.1 degr ...19912026247
prolonged milk residue in two cows after subcutaneous injections of penicillin at an extra-label dose.after cesarian section was done on 2 holstein cows, procaine penicillin g was injected sc at an extra-label-dose. the milk contained inhibitory residue for 21 days (case 1) and 10 days (case 2) after the final penicillin injection. the bacillus stearothermophilus disk assay was used to confirm presence of an inhibitor. analysis to specifically identify the inhibitor substance was not performed. other sources of residues were examined and excluded in these cases. procaine penicillin g was assumed ...19912026539
the cytosolic and glycosomal glyceraldehyde-3-phosphate dehydrogenase from trypanosoma brucei. kinetic properties and comparison with homologous enzymes.the protozoan haemoflagellate trypanosoma brucei has two nad-dependent glyceraldehyde-3-phosphate dehydrogenase isoenzymes, each with a different localization within the cell. one isoenzyme is found in the cytosol, as in other eukaryotes, while the other is found in the glycosome, a microbody-like organelle that fulfils an essential role in glycolysis. the kinetic properties of the purified glycosomal and cytosolic isoenzymes were compared with homologous enzymes from other organisms. both trypa ...19912040304
bsiy i, a novel thermophilic restriction endonuclease that recognizes 5' ccnnnnnnngg 3' and the discovery of a wrongly sequenced site in pacyc177.a new type ii restriction endonuclease designated bsiy i has been purified from a thermophilic soil bacillus stearothermophilus strain. this enzyme recognizes and cleaves the highly degenerate sequence 5' ccnnnnn!nngg 3'. during the identification of the recognition sequence of bsiy i, we discovered that there should be five g nucleotides instead of four at position 1227-1230 of the plasmid pacyc177.19912041772
in vivo transcriptional pattern in the infc operon of bacillus stearothermophilus.by northern blot and primer extension analyses it was shown that in bacillus stearothermophilus the genes infc, rpmi and rplt constitute a single transcriptional unit; the promoter and the transcriptional start-point used in vivo were identified and the half-life of the transcript (1.2 min) was determined. no indication of multiple initiation sites nor of differential stability of different regions of the transcript was found. the results suggest that escherichia coli and b. stearothermophilus h ...19912046659
molecular cloning and nucleotide sequencing of the aspartate racemase gene from lactic acid bacteria streptococcus thermophilus.the gene coding aspartate racemase (ec 5.1.1.13) was cloned from the lactic acid bacteria streptococcus thermophilus iam10064 and expressed efficiently in escherichia coli. the 2.1 kilobase pairs long full length clone had an open reading frame of 729 nucleotides coding for 243 amino acids. the calculated molecular weight of 27,945 agreed well with the apparent molecular weight of 28,000 found in sodium dodecyl sulfate (sds) polyacrylamide gel electrophoresis of the aspartate racemase purified f ...19912054383
cloning and analysis of the nuclear gene for yml33, a protein of the large subunit of the mitochondrial ribosome in saccharomyces cerevisiae.the n-terminal amino acid sequence of a large subunit protein, termed yml33, of the mitochondrial ribosome of the yeast saccharomyces cerevisiae was determined. the data were obtained to synthesize two kinds of oligonucleotide primers, which were used in the polymerase chain reaction to amplify and clone the nuclear gene for this protein. by nucleotide sequencing, the cloned gene, mrp-l33, was found to encode a basic protein of 11 kda with 98 amino acid residues. the protein encoded by this gene ...19912061283
[molecular cloning and structural-functional analysis of the arginine biosynthesis genes of the thermophilic bacterium bacillus stearothermophilus].genes encoding arginine biosynthesis of bacillus stearothermophilus strain 718 were cloned in the mutant arga strain of the escherichia coli k-12. the arg genes were shown to be located on the 3.7 kb dna fragment in the following order: arga--arge--argb. the expression of the arga gene of b. stearothermophilus on the multicopy vehicle is twofold higher in argr- strain of e. coli k-12 than is isogenic argr+ strain. according to hybridization analysis arga genes of b. stearothermophilus and b. sub ...19902074006
thermostable glucokinase from bacillus stearothermophilus and its analytical application. 19902075989
[residues of inhibitory agents in the tissues of slaughter-house animals--comparison of microbiological methods of agar diffusion].three microbiological methods of agar diffusion were compared which are used to detect the residues of inhibitory substances: method using the strain bacillus subtilis atcc 6633 (b.s. 6633), method using the strain bacillus stearothermophilus v. calidolactis c 953 (b. s. v. c. 953), and four-plate method. using the compared methods, minimum inhibitory concentrations were determined for standard solutions of antibiotics and sulphadimidine. inhibitory substances were detected parallely in the samp ...19902087802
nucleotide sequence and cloning in bacillus subtilis of the bacillus stearothermophilus pleiotropic regulatory gene degt.the regulatory gene (degt) from bacillus stearothermophilus nca1503 which enhanced production of extracellular alkaline protease (apr) was cloned in bacillus subtilis with ptb53 as a vector. when b. subtilis mt-2 (npr- [deficiency of neutral protease] apr+) was transformed with the recombinant plasmid, pdt145, the plasmid carrier produced about three times more alkaline protease than did the wild-type strain. in contrast, when b. subtilis db104 (npr- apr-) was used as a host, the transformant wi ...19902104607
vapor-phase hydrogen peroxide as a surface decontaminant and sterilant.the feasibility of utilizing vapor-phase hydrogen peroxide (vphp) as a surface decontaminant and sterilant was evaluated in a centrifuge application. the prototype vphp decontamination system, retrofitted into a beckman l8-m ultracentrifuge, was designed to vaporize a 30% (wt/wt) solution of aqueous hydrogen peroxide continuously injecting and withdrawing vphp in a deep-vacuum flow-through system. vphp cycles of 4, 8, 16, and 32 min were examined for cidal activity against spores of bacillus sub ...19902106287
isolation and molecular genetic characterization of the bacillus subtilis gene (infb) encoding protein synthesis initiation factor 2.western blot (immunoblot) analysis of bacillus subtilis cell extracts detected two proteins that cross-reacted with monospecific polyclonal antibody raised against escherichia coli initiation factor 2 alpha (if2 alpha). subsequent southern blot analysis of b. subtilis genomic dna identified a 1.3-kilobase (kb) hindiii fragment which cross-hybridized with both e. coli and bacillus stearothermophilus if2 gene probes. this dna was cloned from a size-selected b. subtilis plasmid library. the cloned ...19902110148
overproduction of tyrosyl-trna synthetase is toxic to escherichia coli: a genetic analysis.the tyrs genes from escherichia coli and bacillus stearothermophilus were toxic to e. coli when they were carried by plasmids with very high copy numbers (pembl8 and pembl9). we quantified this effect by comparing the efficiencies of plating of e. coli derivatives harboring recombinant plasmids in various experimental conditions. the toxicity was apparent at both 30 and 37 degrees c. it increased with the growth temperature, the strength of the tyrs promoter, and the copy number of the plasmidic ...19902113914
construction and properties of a temperature-sensitive mutation in the gene for the bacteriophage spo1 dna-binding protein tf1.the bacillus subtilis bacteriophage spo1 encodes the dna-binding protein tf1, a homolog of the ubiquitous type ii dna-binding proteins that are components of bacterial chromatin. the known three-dimensional structure of a related protein was used in devising a scheme of site-directed mutagenesis that led to the creation of a temperature-sensitive mutation in the tf1 gene. at the nonpermissive temperature, this mutation disrupted the temporal regulation of viral protein synthesis and processing, ...19902115873
characterization of a thermostable bacillus stearothermophilus alpha-amylase.liquefying-type bacillus stearothermophilus alpha-amylase was characterized. the coding gene was cloned in bacillus subtilis and the enzyme was produced in three different host organisms: b. stearothermophilus, b. subtilis, and escherichia coli. properties of the purified enzyme were similar irrespective of the host. temperature optimum was at 70-80 degrees c and ph optimum at 5.0-6.0. the enzyme was stable for 1 h in the ph range 6.0-7.5 at 80 degrees c. the enzyme was stabilized by ca2+, na+, ...19902119192
glutamyl-trna synthetases of bacillus subtilis 168t and of bacillus stearothermophilus. cloning and sequencing of the gltx genes and comparison with other aminoacyl-trna synthetases.the glutamyl-trna synthetase (glurs) of bacillus subtilis 168t aminoacylates with glutamate its homologous trna(glu) and trna(gln) in vivo and escherichia coli trna(1gln) in vitro (lapointe, j., duplain, l., and proulx, m. (1986) j. bacteriol. 165, 88-93). the gltx gene encoding this enzyme was cloned and sequenced. it encodes a protein of 483 amino acids with a mr of 55,671. alignment of the amino acid sequences of four bacterial glurss (from b. subtilis, bacillus stearothermophilus, e. coli, a ...19902120226
thermal analysis of bacteria by differential scanning calorimetry: relationship of protein denaturation in situ to maximum growth temperature.differential scanning calorimetry (dsc) was used to analyze thermal transitions in two strains of the thermophile bacillus stearothermophilus (atcc 12016 and wat), the mesophile bacillus megaterium and the psychrotroph bacillus psychrophilus. the observed transitions, representing lipid melting and dna and protein unfolding, are compared to the maximum growth temperature (tmax) in each species as a means of identifying critical, thermolabile targets responsible for heat-induced inhibition of gro ...19902121283
growth of bacillus stearothermophilus on glycerol in chemostat culture: expression of an unusual phenotype.bacillus stearothermophilus grew readily on glycerol in carbon-limited chemostat culture and expressed a high carbon conversion efficiency. however, the strain of organism used (probably b. stearothermophilus var. nondiastaticus) proved particularly sensitive to glycerol, both respiration and growth being severely impeded by any surfeit of this compound. sensitivity was found to correlate with an exceptionally high level of expression of glycerol kinase [activities of more than 80 mumol min-1 (m ...19902121901
purification and properties of thermostable xylanase and beta-xylosidase produced by a newly isolated bacillus stearothermophilus strain.we isolated a thermophilic bacterium that produces both xylanase and beta-xylosidase. based on taxonomical research, this bacterium was identified as bacillus stearothermophilus. each extracellular enzyme was separated by hydrophobic chromatography by using a toyopearl hw-65 column, followed by gel filtration with a sephacryl s-200 column. each enzyme in the culture was further purified to homogeneity (62-fold for xylanase and 72-fold for beta-xylosidase) by using a fast protein liquid chromatog ...19902123854
the thermostability of dna-binding protein hu from bacilli.the primary and tertiary structures of dna-binding protein hu from bacillus stearothermophilus are already known. the primary structure has been previously determined for hu from the closely related b. globigii and the determinations of the sequences from b. caldolyticus and b. subtilis are described here. these bacteria have optimum growth temperatures of greater than 70 degrees c (b. caldolyticus), 65 degrees c (b. stearothermophilus), 37 degrees c (b. subtilis) and 30 degrees c (b. globigii). ...19902127103
heat sterilization of bioindicators in propylene glycol and propylene glycol-water mixtures: arrhenius equation, thermodynamic data, and z values.our interest in calculating the thermodynamic data by means of the arrhenius equation was based on two observation: (a) the thermal death time increases considerably when the bioindicators bacillus subtilis var. niger and bacillus stearothermophilus are sterilized in nonaqueous hydrophilic solutions as found in propylene glycol (pg) with low water concentrations; and (b) the inactivation kinetics of bac. stearothermophilus does not follow a first-order reaction. the frequency factor a and the en ...19902128895
dynamic ultraviolet sterilization of different implant types.this paper investigates the use of the dynamic ultraviolet sterilization process with various dental implants, stainless steel orthopedic cortical bone screws, and polysulfone polymer healing caps. these biomaterials were inoculated with the spores of bacillus subtilis and bacillus stearothermophilus. they were then exposed to dynamic ultraviolet radiation in the chamber of a bud ultraviolet device. samples were incubated in trypticase soy broth at 37 degrees c and 56 degrees c, and they were su ...19902133336
presence of the bacterial hemoglobin gene improves alpha-amylase production of a recombinant escherichia coli strain.a recombinant plasmid (pmk57) was constructed by cloning the bacillus stearothermophilus alpha-amylase gene into puc8; plasmid pmk79 was then derived from pmk57 by inserting the bacterial (vitreoscilla) hemoglobin gene into the latter plasmid. both pmk57 and pmk79 were transformed into escherichia coli strain jm 103 to make strains mk57 and mk79, respectively. both mk57 and mk79 produced alpha-amylase and mk79 produced hemoglobin. mk79 outgrew mk57 in shake flasks in lb medium, the advantage of ...19902136531
tetramer-dimer conversion of phosphofructokinase from thermus thermophilus induced by its allosteric effectors.phosphofructokinase (pfkase) was purified from an extreme thermophile. thermus thermophilus. allosteric natures of t. thermophilus pfkase is similar to those of bacillus stearothermophilus pfkase, that is, hyperbolic plots of the activity versus concentration of fructose 6-phosphate (f6p) were changed into a sigmoidal shape by the addition of phosphoenolpyruvate (pep), while further addition of adp caused it to revert to a hyperbolic shape. the native t. thermophilus pfkase has an mr of 148,000 ...19902146397
the rabbit muscle phosphofructokinase gene: cdna cloning and sequencing.a partial cdna for rabbit muscle phosphofructokinase (rm-pfk) has been cloned and sequenced. the nucleotide sequence of the cdna agrees with the previously determined rm-pfk genomic sequence. in addition, the amino acid sequence deduced from the cdna is nearly identical to the rm-pfk sequence previously determined by peptide analysis. a significant degree of homology exists when the amino acid sequence of rm-pfk is compared with the sequences of bacillus stearothermophilus pfk or escherichia col ...19902147292
stereochemistry and lifetime of the gtp hydrolysis intermediate at the active site of elongation factor tu from bacillus stearothermophilus as inferred from the 17o-55mn superhyperfine interaction.electron paramagnetic resonance spectroscopy has been used to obtain information on the structure and stability of the products of gtp cleavage at the active site of elongation factor tu (ef-tu) from bacillus stearothermophilus. using stereospecifically labelled (sp)-(rp)-[beta-17o]gtp (prepared by modification of a previously published procedure which is now also suitable for guanine nucleotides), it was found that only one of the two possible diastereomers (sp) led to detectable line-broadenin ...19902156700
purification and characterization of thermostable beta-mannanase and alpha-galactosidase from bacillus stearothermophilus.bacillus stearothermophilus secretes beta-mannanase and alpha-galactosidase enzymatic activities capable of hydrolyzing galactomannan substrates. expression of the hemicellulase activities in the presence of locust bean gum was sequential, with mannanase activity preceding expression of alpha-galactosidase activity. the hemicellulase activities were purified to homogeneity by a combination of ammonium sulfate fractionation, gel filtration, hydrophobic interaction chromatography, and ion-exchange ...19902176449
overexpression of the methanococcal ribosomal protein l12 in escherichia coli and its incorporation into halobacterial 50 s subunits yielding active ribosomes.the gene for the ribosomal l12 protein from the archaebacterium methanococcus vannielii was cloned into the expression vector pkk223-3. the protein was overexpressed and remained stable in escherichia coli xl1 cells. purification yielded a protein with the same amino acid composition and sequence as in methanococcus but it was acetylated at the n terminus as in the case with the homologous protein of e. coli. the in vivo incorporation of the overexpressed protein into the e. coli ribosomes was n ...19902180948
evidence that chemical modification of a positively charged residue at position 189 causes the loss of catalytic activity of iron-containing and manganese-containing superoxide dismutases.the escherichia coli, bacillus stearothermophilus, and human manganese-containing superoxide dismutases (mnsods) and the e. coli iron-containing superoxide dismutase (fesod) are extensively inactivated by treatment with phenylglyoxal, an arginine-specific reagent. arg-189, the only conserved arginine in the primary sequences of these four enzymes, is also conserved in the three additional fesods and five of the six additional mnsods sequenced to date. the only exception is saccharomyces cerevisi ...19902186704
thermostable alanine racemase. its structural stability.the gene encoding thermostable alanine recemase from bacillus stearothermophilus was cloned and expressed in e. coli. the enzyme was purified to homogeneity from cell extracts of e. coli carrying a plasmid designated picr4. the alanine racemase gene sequenced was found to contain an open reading frame of 1158 nucleotides. the molecular weight of the enzyme subunit was estimated to be 43,341. the alpha-helical and beta-structure contents were calculated to be about 34 and 26%, respectively, from ...19902192620
expression in escherichia coli of a sub-gene encoding the lipoyl domain of the pyruvate dehydrogenase complex of bacillus stearothermophilus.a sub-gene encoding the lipoyl domain (residues 1-85) of the lipoate acetyltransferase chain of the pyruvate dehydrogenase complex of bacillus stearothermophilus was over-expressed in escherichia coli. approx. 80% of the domain was unlipoylated but most of the remainder was correctly lipoylated on lys-42 and could be reductively acetylated by the b stearothermophilus enzyme complex. a small proportion (approx. 4%) of the domain carried an aberrant substituent, possibly an octanoyl group, on lys- ...19902192914
amino acid sequences of the ribosomal proteins hl30 and hmal5 from the archaebacterium halobacterium marismortui.the complete amino acid sequences of the ribosomal proteins hl30 and hmal5 from the archaebacterium halobacterium marismortui were determined. protein hl30 was found to be acetylated at its n-terminal amino acid and shows homology to the eukaryotic ribosomal proteins yl34 from yeast and rl31 from rat. protein hmal5 was homologous to the protein l5 from escherichia coli and bacillus stearothermophilus as well as to yl16 from yeast. hmal5 shows more similarities to its eukaryotic counterpart than ...19902198942
cloning and sequence analysis of the genes encoding the alpha and beta subunits of the e1 component of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.a 4175-bp ecori fragment of dna that encodes the alpha and beta chains of the pyruvate dehydrogenase (lipoamide) component (e1) of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been cloned in escherichia coli. its nucleotide sequence was determined. open reading frames (pdha, pdhb) corresponding to the e1 alpha subunit (368 amino acids, mr 41,312, without the initiating methionine residue) and e1 beta subunit (324 amino acids, mr 35,306, without the initiating ...19902200674
cloning and nucleotide sequences of the bacillus stearothermophilus neutral protease gene and its transcriptional activator gene.both the neutral protease gene (nprs) and its transcriptional activator gene (npra) from bacillus stearothermophilus telne were cloned in bacillus subtilis by using ptb53 as a vector plasmid. the presence of the npra gene enhanced protease synthesis by about fivefold. the nucleotide sequences of nprs and its flanking regions were determined. nprs was composed of 1,653 base pairs and 551 amino acid residues. a shine-dalgarno (sd) sequence was found 9 bases upstream from the translation start site ...19902203733
expression of the copy dna for human a4 and b4 l-lactate dehydrogenases in escherichia coli.the human ldh-a and ldh-b cdnas, containing the coding regions for the l-lactate dehydrogenase a4 (m) and b4 (h) polypeptides respectively have been cloned into escherichia coli to place the cdnas under the control of hybrid e. coli/bacillus stearothermophilus transcriptional and translational signals. human a4- and b4-isoenzymes are produced in e. coli cells harbouring the expression plasmids phldha22 and phldhb10 at levels of 6.5 and 1.5% of the soluble protein of the cell, respectively. the t ...19902205297
nucleotide sequences of bacillus stearothermophilus ribosomal protein genes: part of the ribosomal s10 operon.restriction fragments from bacillus stearothermophilus chromosomal dna were cross-hybridized with the escherichia coli ribosomal protein l2 gene rplb. a 2-kb ecori fragment which showed cross-hybridization was cloned into the m13 phage and sequenced by the dideoxy chain-terminating method. comparison of the deduced amino-acid sequences with the corresponding sequences of e. coli ribosomal proteins showed that this fragment contains the region encoding the c-terminus of l2, the genes encoding s19 ...19902222862
probing the coenzyme specificity of glyceraldehyde-3-phosphate dehydrogenases by site-directed mutagenesis.by combining our knowledge of the crystal structure of the glycolytic nad-dependent glyceraldehyde-3-phosphate dehydrogenase (gapdh) and the sequence of the photosynthetic nadp-dependent gapdh of the chloroplast, two particular amino acid residues were predicted as the principal determinants of differing coenzyme specificity. by use of site-directed mutagenesis, the amino acids leu 187 and pro 188 of gapdh from bacillus stearothermophilus have been replaced with ala 187 and ser 188, which occur ...19902223764
a comparative study of ribosomal and dna binding protein ii from two thermophilic bacteria, bacillus caldolyticus strain ep 00275 and bacillus stearothermophilus.ribosomal and dna binding proteins (dna bp ii) from an extreme thermophilic bacterium, b. caldolyticus strain ep 00275, were investigated for stability and crystallization and compared to the homologous proteins from b. stearothermophilus. two-dimensional gel electrophoresis of both types of proteins, the amino acid composition and the sequences of some of the peptides of dna bp ii revealed a close relationship between each other. the physico-chemical characteristics of dna bp ii were similar bu ...19902226856
use of a hapten specific anti-dansyl antibody for the localization of ribosomal proteins by immuno electron microscopy.the fluorescent reagent dansyl chloride has been used as an immunological marker for the electron microscopic localization of ribosomal proteins on the surface of 50s ribosomal subunits. the proteins bstl1 from bacillus stearothermophilus and ecol1 from escherichia coli were dansylated to various degrees and reconstituted into the l1-deficient e. coli 50s subunits from mutant mv17-10. using antibodies specific to dansyl chloride, both proteins were mapped at the lateral protuberance near the pep ...19902242000
[construction of promoter probe vector pfdc4 and gene expression vector pfdc11 with high transformation efficiency in bacillus stearothermophilus].a 0.5kb fragment from sau3a-digested total dna of bacillus stearothermophilus cu21 was cloned with the vector ppgv5, a derivative of ppl703. the insertion of this fragment can activate the expression of the promoteless cat-86 gene on the cloning vector in both b. stearothermophilus and bacillus subtilis hosts. when the 0.54 kb fragment is present in ppgv5 in either orientation, the transformation efficiency of the plasmid is increased about 10(3) to 10(4) fold in cu21 protoplasts. southern hybri ...19902242283
role of anionic lipid in bacterial membranes.the major phospholipids of bacillus stearothermophilus are phosphatidylethanolamine (pe), phosphatidylglycerol (pg), and cardiolipin (cl). under the growth conditions used in this study the concentration of anionic lipid (pg + cl) was determined by the ph of the culture medium. cells grown in a complex medium at ph 5.8, 7.0, and 8.0 contained 17, 29 and 36 nmol of anionic (pg + cl) lipid/mg cell (dry weight). the concentration of the zwitterionic lipid phosphatidylethanolamine (pe) was 17-20 nmo ...19902248965
[a novel type of phase variation regarding integrated and free states of plasmid pfdx163 in bacillus stearothermophilus cu21].pfdx1 is a recombinant plasmid which carries a foreign gene xyle. by selecting for kanamycin-resistant mutants of bacillus stearothermophilus cu21(pfdx1) at higher temperature, a variant strain cu21-163 was obtained. this strain harbors a mutant plasmid pfdx163, which was formed by insertion of a 2.0kb h-fragment from the cu21 genome onto the plasmid pfdx1. pfdx163 was supposed to be integrated into the cu21 chromosome via homologous recombination of h-fragments. the cu21-163 strain consists of ...19902252599
cloning and sequence analysis of the genes encoding the dihydrolipoamide acetyltransferase and dihydrolipoamide dehydrogenase components of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.a 2641-bp ecori fragment of dna that encodes the c-terminal part of the dihydrolipoyl acetyltransferase (e2) component and the dihydrolipoamide dehydrogenase (e3) component of the pyruvate dehydrogenase complex of bacillus stearothermophilus has been cloned in escherichia coli. its nucleotide sequence was determined. a 705-bp truncated open reading frame was located at the 5'end of the insert which, together with the 588-bp truncated open reading frame at the 3' end of another ecori fragment of ...19902253629
isolation and characterization of bacillus stearothermophilus 30s and 50s ribosomal protein mutations.bacillus stearothermophilus mutations which confer resistance to or dependence on a variety of ribosome-targeted antibiotics have been isolated. many of these mutations produce ribosomal proteins with altered mobilities in a two-dimensional gel electrophoresis system. this collection of altered thermophilic ribosomal proteins will be useful in examining ribosomal structure and function.19902254291
proteolysis of bacillus stearothermophilus if2 and specific protection by gtp.translation initiation factor if2 from bacillus stearothermophilus (741 amino acids, mr = 82,043) was subjected to trypsinolysis alone or in the presence of gtp. following electroblotting and automated amino acid sequencing of the resulting peptides, the location and the sequential order of the main cleavage sites were identified. trypsinolysis of if2 ultimately generates two compact domains: a 24.5 kda c-terminal fragment and a 40 kda g-fragment which is obtained only in the presence of gtp whi ...19902265694
in vivo genetic engineering: homologous recombination as a tool for plasmid construction.this paper describes a novel method for creating exact dna fusions between any two points in a plasmid carried in bacillus subtilis. it exploits the homologous in vivo recombination between directly repeated sequences that can be established by insertion of a synthetic oligodeoxyribonucleotide. the method was used to enhance the productivity in b. subtilis of a cloned alpha-amylase (amy)-encoding gene originating from bacillus stearothermophilus. thus, an exact fusion between nucleotide sequence ...19902265757
designs for a broad substrate specificity keto acid dehydrogenase.variations have been made to the structure of the nicotinamide adenine dinucleotide (nad) dependent l-lactate dehydrogenase from bacillus stearothermophilus at regions of the enzyme that we believe determine specificity toward different alpha-hydroxy acids (rchohcoo-, r = ch3, c2h5, etc.). two regions of ldh that border the active site (but are not involved in the catalytic reaction) were altered in order to accommodate substrates with hydrophobic side chains larger than that of the naturally pr ...19902271542
identification of a reversible structural transition in the metal-depleted glycerol dehydrogenase from bacillus stearothermophilus.evidence is presented to demonstrate that the zn2+ -depleted, inactive form of the glycerol dehydrogenase from bacillus stearothermophilus exists in one of two possible conformations in equilibrium, the position of which is temperature sensitive. the conformation of the metal-depleted enzyme favoured by higher temperatures (20-40 degrees c) is able to bind zn2+ and regain catalytic activity, whereas that favoured at lower temperatures (0-10 degrees c) is unable to bind metal ions and is thus ina ...19902294019
genetic map of the bacillus stearothermophilus nub36 chromosome.a circular genetic map of bacillus stearothermophilus nub36 was constructed by transduction with bacteriophage tp-42c and protoplast fusion. sixty-four genes were tentatively assigned a cognate bacillus subtilis gene based on growth response to intermediates or end products of metabolism, cross-feeding, accumulation of intermediates, or their relative order in a linkage group. although the relative position of many genes on the bacillus stearothermophilus and bacillus subtilis genetic map appear ...19902298700
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