| crystals of wild-type, mutated, derivatized and complexed 50 s ribosomal subunits from bacillus stearothermophilus suitable for x-ray analysis. | three-dimensional single crystals of wild-type and mutated 50 s ribosomal subunits from bacillus stearothermophilus, as well as crystals of reconstituted subunits containing heavy-atom clusters and complexes of these subunits with trna and a short nascent polypeptide chain, were grown from polyethylene glycol in the presence of salts at low concentrations. within experimental error, all these crystals are isomorphous, packed with monoclinic symmetry (c2) in unit cells of a = 300 a, b = 546 a, c ... | 1989 | 2467005 |
| structural and kinetic bases for the recognition of trnatyr by tyrosyl-trna synthetase. | the aminoacylation of transfer rna is a key step of translation since it relates amino acids to anticodons. to understand how the tyrosyl-trna synthetase (tyrts) from bacillus stearothermophilus recognizes trna(tyr), we constructed 14 new mutant tyrts by site-directed mutagenesis, determined their kinetic properties and used these and previous data to construct a detailed structural model of the complex between tyrts and the acceptor arm of trna(tyr). in the model arg207, lys208, asn 146 and glu ... | 1989 | 2467006 |
| nucleotide sequence of the neopullulanase gene from bacillus stearothermophilus. | the gene (nplt) for a new type of pullulan-hydrolysing enzyme, neopullulanase, from bacillus stearothermophilus trs40 was sequenced. the dna sequence revealed only one large open reading frame, composed of 1764 bases and 588 amino acid residues (mr 69144). although the thermostable neopullulanase contained eight cysteine residues, they did not provide conformational stability by disulphide bonds. a comparison was made of the amino acid sequences of alpha-amylase, neopullulanase, isoamylase, pull ... | 1989 | 2482332 |
| expression of the insecticidal protein gene from bacillus thuringiensis subsp. aizawai in bacillus subtilis and in the thermophile bacillus stearothermophilus by using the alpha-amylase promoter of the thermophile. | expression of the insecticidal protein gene from bacillus thuringiensis subsp. aizawai ipl7 in b. subtilis mi113 and b. stearothermophilus sic1 was examined. production of the protein (130 kilodaltons [kda]) was analyzed by its reaction with antibody against the insecticidal proteins of the parental b. thuringiensis. when the original gene containing its own promoter was subcloned in b. subtilis, only a small amount of the protein was produced. therefore, both the promoter for the b. stearotherm ... | 1989 | 2482704 |
| isolation and characterisation of the glycerol dehydrogenase from bacillus stearothermophilus. | a protocol for the rapid purification of the glycerol dehydrogenase (glycerol: nad+ 2-oxidoreductase, ec 1.1.1.6) from the thermophile bacillus stearothermophilus has been developed using a combination of chromatographic techniques including affinity chromatography on a sepharose-immobilised triazine dye (procion red, he3b, ici). substrate specificity has been examined and km values determined. the protein has been shown to have an oligomeric mr of approx. 180,000 and consists of four identical ... | 1989 | 2493267 |
| viable starter culture, beta-galactosidase activity, and lactose in duodenum after yogurt ingestion in lactase-deficient humans. | ten lactose malabsorbers were intubated and given fresh or heated yogurt to which polyethylene-glycol (peg) and spores of bacillus stearothermophilus (sbs) had been added as internal standards. in duodenal samples taken after fresh yogurt ingestion, viable starter culture was detected for 60 min in 6 of 7 subjects and the ratio of microbial beta-galactosidase activity to sbs remained similar during this period to its value in the preingested yogurt. in the two groups ingesting fresh and heated y ... | 1989 | 2497633 |
| thermophilic bacillus sp. that shows the denitrification phenotype of pseudomonas aeruginosa. | a thermophilic bacillus sp. of marine origin was observed to grow anaerobically on nitrite, nitrous oxide (n2o) in the presence of nitrite, and n2o alone for a few hours after exhaustion of nitrite. this represents the second example of a denitrification phenotype originally observed to occur with pseudomonas aeruginosa. | 1989 | 2499254 |
| on the effect on specificity of thr246----gly mutation in l-lactate dehydrogenase of bacillus sterothermophilus. | the function of the amino acid thr246 in l-lactate dehydrogenase from bacillus stearothermophilus has been investigated by site-directed replacement with glycine. kinetic experiments with a number of 2-oxo acids showed strongly reduced activity for the mutated enzyme. however, the mutant enzyme shows a relative preference for the large hydrophobic sidechains of alpha-keto acids and an even higher specific activity than the wild-type lactate dehydrogenase for the polar oxaloacetate substrate. gra ... | 1989 | 2499337 |
| one-step affinity purification of bacillus neutral proteases using bacitracin-silica. | a purification procedure for neutral proteases from bacilli is described, in which bacitracin-silica was used as affinity medium. this enabled a one-step purification of the proteases directly from culture supernatant. since neutral proteases are extremely sensitive towards autodigestion, conditions were chosen such, that autodigestion was largely prevented. isopropanol appeared to be useful in both eluting the enzymes from the affinity medium, and inhibiting enzymatic activity during this step. ... | 1989 | 2499614 |
| structural comparison of 26s rrna-binding ribosomal protein l25 from two different yeast strains and the equivalent proteins from three eubacteria and two chloroplasts. | the sequences of saccharomyces carlsbergensis ribosomal protein (r-protein) sl25* and its equivalents from candida utilis (cl25), escherichia coli (el23), bacillus stearothermophilus (bl23), mycoplasma capricolum (ml23), marchantia polymorpha chloroplasts (mcpl23), and nicotiana tabacum chloroplasts (ncpl23) were examined using a computer program that evaluates the extent of sequence similarity by calculating correlation coefficients for each pair of residues in two proteins from a number of phy ... | 1989 | 2501503 |
| site-directed mutagenesis of glyceraldehyde-3-phosphate dehydrogenase reveals the role of residue ser148. | a mutant of bacillus stearothermophilus d-glyceraldehyde-3-phosphate dehydrogenase, ser148----ala, was produced by oligonucleotide-directed mutagenesis. the study of the catalytic properties of this mutant has shown that this mutation significantly affects the michaelis constant of inorganic phosphate and to a lesser extent that of 1,3-diphosphoglycerate and d-glyceraldehyde-3-phosphate. this result is consistent with model-building studies which show that, for the phosphorylation step of cataly ... | 1989 | 2501780 |
| modification of bacillus subtilis elongation factor tu by n-tosyl-l-phenylalanyl chloromethane abolishes its ability to interact with the 3'-terminal polynucleotide structure but not with the acyl bond in aminoacyl-trna. | modification of b. subtilis ef-tu by n-tosyl-l-phenylalanyl chloromethane destroyed its ability to promote protein synthesis and resulted in selective dissociation of the two binding activities of the protein for aminoacyl-trna. the modified ef-tu was completely ineffective in the protection of the 3'-terminal cca structure of trna against pancreatic ribonuclease, while remaining almost fully active in the protection of the ester bond between the 3'-terminal adenosine and the amino acid residue ... | 1989 | 2502433 |
| overproduction of thermostable leucine dehydrogenase of bacillus stearothermophilus and its one-step purification from recombinant cells of escherichia coli. | we have cloned the leucine dehydrogenase (ec 1.4.1.9) gene from a thermophile, bacillus stearothermophilus, into escherichia coli mv1184 with a vector plasmid, puc119. the cloned cells produced a large amount of the thermostable enzyme, which corresponds to about 60% of the total soluble protein. the enzyme was purified to more than 95% homogeneity by only one step, heat treatment of the cell-extracts, with an average yield of 75 mg/g of wet cells (obtained from 100 ml of the culture). | 1989 | 2503013 |
| structure of tyrosyl-trna synthetase refined at 2.3 a resolution. interaction of the enzyme with the tyrosyl adenylate intermediate. | the crystal structure of tyrosyl-trna synthetase (ec 6.1.1.1) from bacillus stearothermophilus has been refined to a crystallographic r-factor of 22.6% at 2.3 a resolution using a restrained least-squares procedure. in the final model the root-mean-square deviation from ideality for bond distances is 0.018 a and for angle distances is 0.044 a. each monomer consists of three domains: an alpha/beta domain (residues 1 to 220) containing a six-stranded beta-sheet, an alpha-helical domain (248 to 318 ... | 1989 | 2504923 |
| isocitrate dehydrogenase from thermophilic and mesophilic bacteria. isolation and some characteristics. | 1. simple methods incorporating the principle of selective enzyme elution from a triazinyl dye adsorbent with a mixture of nadp+ and isocitrate are described for isolating nadp+-linked isocitrate dehydrogenase in pure state from several mesophilic and thermophilic bacteria. 2. several characteristics of the isocitrate dehydrogenases have been examined, viz. molecular size, amino acid composition including the content of sulphydryl groups, thermostability and structural homology by the criterion ... | 1989 | 2515075 |
| site-directed mutagenesis in bacillus stearothermophilus fructose-6-phosphate 1-kinase. mutation at the substrate-binding site affects allosteric behavior. | arg252 of fructose-6-phosphate 1-kinase (pfk) from bacillus stearothermophilus has been proposed to be involved in the binding of the substrate fru-6-p. we demonstrate here that mutation of this residue to alanine converts the enzyme to a form with characteristics similar to those of its allosterically tight form. the mutant enzyme exhibits a high affinity for its inhibitor phosphoenolpyruvate (a 68-fold difference compared to wild type) and a dramatically decreased fru-6-p affinity (1500-fold i ... | 1989 | 2521215 |
| complete nucleotide sequences of two phosphoglucoisomerase isozymes from bacillus stearothermophilus. | | 1989 | 2532320 |
| characterization and application of a thermostable primary transport system: cytochrome-c oxidase from bacillus stearothermophilus. | cytochrome-c oxidase from bacillus stearothermophilus has been purified to homogeneity by detergent extraction followed by deae-cellulose, hydroxyapatite- and gel-filtration chromatography. the enzyme is a typical cytochrome-aa3-type oxidase which binds carbon monoxide and is sensitive to classical oxidase inhibitors like cyanide and azide. the purified enzyme is composed of three different subunits (57, 37 and 22 kda). the subunit with intermediate molecular mass contains a covalently attached ... | 1989 | 2536327 |
| mechanism of l-glutamate transport in membrane vesicles from bacillus stearothermophilus. | in the presence of electrochemical energy, several branched-chain neutral and acidic amino acids were found to accumulate in membrane vesicles of bacillus stearothermophilus. the membrane vesicles contained a stereo-specific transport system for the acidic amino acids l-glutamate and l-aspartate, which could not translocate their respective amines, l-glutamine and l-asparagine. the transport system was thermostable (ti = 70 degrees c) and showed highest activities at elevated temperatures (60 to ... | 1989 | 2563364 |
| the effect of air on the moist-heat resistance of bacillus stearothermophilus spores. | the presence of air in an autoclave chamber and load is generally considered to reduce the lethal effect of the process on bacterial spores. in this study, the heat inactivation of spores of bacillus stearothermophilus (ncib 8224) was measured in the presence and absence of air at 100% relative humidity. the results confirm that air significantly enhances the lethality of moist-heat on this strain of b. stearothermophilus. | 1989 | 2575633 |
| isolation and identification of restriction endonuclease bseai. | | 1989 | 2587237 |
| relationship between the digestive microflora of the newborn and maternal microflora in rodents as evidenced by a transit marker. | the cutaneous and digestive microflora of the dam, constitute the main sources of bacteria likely to colonize the digestive tract of the newborn. our objective was to study the conditions (delay, level) in relation to the maternal microflora. spores of bacillus stearothermophilus were used as a tracer. they were spread on the mother's udder or given per os to the mother. in our study, we used holoxenic rats and axenic mice. the tracer was then followed in newborn rats and newborn mice. transfer ... | 1989 | 2619205 |
| protoplast transformation of bacillus stearothermophilus nub36 by plasmid dna. | an efficient protoplast transformation system was established for bacillus stearothermophilus nub3621 using thermophilic plasmid ptht15 tcr (4.5 kb) and mesophilic plasmid plw05 cmr (3 kb), a spontaneous deletion derivative of ppl401 cmr kmr. the efficiency of transformation of nub3621 with plw05 and ptht15 was 2 x 10(7) to 4 x 10(8) transformants per micrograms dna. the transformation frequency (transformants per regenerant) was 0.5 to 1.0. chloramphenicol-resistant and tetracycline-resistant t ... | 1989 | 2621450 |
| [isolation, purification and properties of restrictase and methylase bstn1 from bacillus stearothermophilus]. | restriction-methylation enzymes bstn1 from bacillus stearothermophilus were isolated and purified. these enzymes are related to a new class of restriction-methylation enzymes of the second type, whose modifying component is n4-cytosine-dna-methylase. both enzymes recognize the dna sequence cc(a/t)gg. restrictase bstn1 is a protein made up of one subunit with a molecular mass of 25 kda. the molecular mass of native dna-methylase bstn1 is about 55 kda. the temperature optima for restrictase and me ... | 1989 | 2627557 |
| structure and properties of malic enzyme from bacillus stearothermophilus. | the malic enzyme (ec 1.1.1.38) gene of bacillus stearothermophilus was cloned in escherichia coli, and the enzyme was purified to homogeneity from the e. coli clone. in addition to the nad(p)-dependent oxidative decarboxylation of l-malate, the enzyme catalyzes the decarboxylation of oxalacetate. the enzyme is a tetramer of mr 200,000 consisting of four identical subunits of mr 50,000. the ph optima for malate oxidation and pyruvate reduction are 8.0 and 6.0, respectively; and the optimum temper ... | 1989 | 2644282 |
| the complete amino acid sequence of ribosomal protein s18 from the moderate thermophile bacillus stearothermophilus. | the amino acid sequence of ribosomal protein s18 from bacillus stearothermophilus has been completely determined by automated sequence analysis of the intact protein as well as of peptides derived from digestion with staphylococcus aureus protease at ph 4.0 and cleavage with cyanogen bromide. the carboxy-terminal region was verified by both amino acid analyses of chymotryptic peptides and by mass spectrometry from the terminal region. the protein contains 77 amino acid residues and has an mr of ... | 1989 | 2647521 |
| cloning, sequence analysis and over-expression of the gene for the class ii fructose 1,6-bisphosphate aldolase of escherichia coli. | nucleotide sequence analysis of the escherichia coli chromosomal dna inserted in the plasmid plc33-5 of the clarke and carbon library [clarke & carbon (1976) cell 9, 91-99] revealed the existence of the gene, fda, encoding the class ii (metal-dependent) fructose 1,6-bisphosphate aldolase of e. coli. the primary structure of the polypeptide chain inferred from the dna sequence of the fda gene comprises 359 amino acids, including the initiating methionine residue, from which an mr of 39,146 could ... | 1989 | 2649077 |
| nucleotide sequence determination of the dna region coding for bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase and of the flanking dna regions required for its expression in escherichia coli. | the complete nucleotide sequence of a 3541-base pairs (bp) dna fragment from bacillus stearothermophilus able to complement an escherichia coli glyceraldehyde-3-phosphate-dehydrogenase (gapdh) mutant (gapd-) has been determined. the b. stearothermophilus gap gene consists of a 1005-bp open reading frame commencing with an atg start codon and ending with a taa stop codon. upstream from the start codon is a strong shine-dalgarno sequence typical of gram-positive bacteria. only one putative rna pol ... | 1989 | 2656407 |
| probing the substrate-binding sites of aminoacyl-trna synthetases with the procion dye green he-4bd. | a reactive bis-dichloro derivative of the procion dye green he-4bd was shown to inactivate irreversibly methionyl-trna synthetase (mts) from escherichia coli and also tryptophyl-trna synthetase (wts) and tyrosyl-trna synthetase (yts) from bacillus stearothermophilus at ph 8.5 and 37 degrees c. at a 5-fold excess of reactive dye over enzyme subunit concentration mts was quantitatively inactivated within 20 min in the atp/pyrophosphate exchange assay, whereas wts and yts show an 80% loss of activi ... | 1989 | 2658972 |
| (1-aminoethyl)boronic acid: a novel inhibitor for bacillus stearothermophilus alanine racemase and salmonella typhimurium d-alanine:d-alanine ligase (adp-forming). | (1-aminoethyl)boronic acid (ala-b), an analogue of alanine in which a boronic acid group replaces the carboxyl group, has been synthesized and found to inhibit the first two enzymes, alanine racemase (from bacillus stearothermophilus, ec 5.1.1.1) and d-alanine:d-alanine ligase (adp-forming) (from salmonella typhimurium, ec 6.3.2.4), of the d-alanine branch of bacterial peptidoglycan biosynthesis. in both cases, time-dependent, slow binding inhibition is observed due to the generation of long-liv ... | 1989 | 2663072 |
| replacement of potassium ions by ammonium ions in different micro-organisms grown in potassium-limited chemostat culture. | the biomass concentration extant in potassium-limited cultures of either klebsiella pneumoniae or bacillus stearothermophilus (when growing at a fixed temperature and dilution rate in a glucose/ammonium salts medium) increased progressively as the medium ph value was raised step-wise from 7.0 to 8.5. because the macromolecular composition of the organisms did not vary significantly, this increase in biomass could not be attributed to an accumulation of storage-type polymers but appeared to refle ... | 1989 | 2669673 |
| cloning, nucleotide sequence, and expression in escherichia coli of the bacillus stearothermophilus peroxidase gene (pera). | the gene encoding a thermostable peroxidase was cloned from the chromosomal dna of bacillus stearothermophilus iam11001 in escherichia coli. the nucleotide sequence of the 3.1-kilobase ecori fragment containing the peroxidase gene (pera) and its flanking region was determined. a 2,193-base-pair open reading frame encoding a peroxidase of 731 amino acid residues (mr, 82,963) was observed. a shine-dalgarno sequence was found 9 base pairs upstream from the translational starting site. the deduced a ... | 1989 | 2670897 |
| addition of a methyl group changes both the catalytic velocity and thermostability of the neutral protease from bacillus stearothermophilus. | specific activity was compared between wild-type (wt) neutral protease from bacillus stearothermophilus and mutant protease (m1; gly144 replaced by ala144) with enhanced thermostability. when casein was used as a substrate, m1 showed 1.5-times higher specific activity than that of wt. in contrast, the specific activities of m1 for soluble reduced lysozyme and insulin b chain were lower than those of wt by 17.2 and 13.2%, respectively. after digestion of the insulin a chain by these enzymes, the ... | 1989 | 2673840 |
| nucleotide sequences of genes encoding heat-stable and heat-labile glyceraldehyde-3-phosphate dehydrogenases; amino acid sequence and protein thermostability. | the structural gene (gapst) encoding glyceraldehyde-3-phosphate dehydrogenase (gpdh; ec 1.2.1.12) from bacillus stearothermophilus has been cloned in escherichia coli using plasmid pbr322 as a vector; the homologous gene (gapco) from bacillus coagulans was cloned from a phage lambda library. expression of the cloned gap genes revealed that, as in the wild-type (wt) organisms, the gpdh from b. stearothermophilus (gpdh-st) was intrinsically heat stable (hs) and that from b. coagulans (gpdh-co) hea ... | 1989 | 2684782 |
| 2-oxo acid dehydrogenase multienzyme complexes: domains, dynamics, and design. | | 1989 | 2699393 |
| 4-chloroacetylpyridine adenine dinucleotide. a highly reactive and chromophoric affinity label of glyceraldehyde-3-phosphate dehydrogenase from sturgeon. | the analogue of nad+, 4-chloroacetylpyridine-adenine dinucleotide (clac4pdad+), inactivated the glyceraldehyde-3-phosphate dehydrogenase from sturgeon at a high rate. an affinity labeling was shown to occur with clac4pdad+. the mononucleotide 4-chloroacetylpyridine 1-beta-d-ribose 5'-phosphate (clac4pdmn+) reacted with the enzyme in a second-order reaction whose rate was much smaller than that calculated for clac4pdad+ taken as a second-order rate reagent. the rate of the reaction of clac4pdad+ ... | 1989 | 2714279 |
| isolation of phenol-degrading bacillus stearothermophilus and partial characterization of the phenol hydroxylase. | bacillus stearothermophilus br219, isolated from river sediment, degraded phenol at levels to 15 mm at a rate of 0.85 mumol/h (4 x 10(6) cells). the solubilized phenol hydroxylase was nadh dependent, exhibited a 55 degrees c temperature optimum for activity, and was not inhibited by 0.5 mm phenol. | 1989 | 2719481 |
| a study of the use of autoclave bags in non-vacuum autoclaves. | two types of autoclave bags have been tested in five commonly used dental, non-vacuum autoclaves. the bags did not prevent the sterilization of instruments provided that most of the air was removed and the bags of instruments were not stacked. the use of a sterilization indicator in the bag is mandatory. | 1989 | 2768625 |
| cloning and characterization of a gene cluster from bacillus stearothermophilus comprising infc, rpmi and rplt. | using two synthetic deoxyribonucleotide probes encoding segments of the primary structure of initiation factor if3 from bacillus stearothermophilus, we identified and cloned a segment of dna which carries the infc gene. as in escherichia coli, the infc gene begins with the unusual initiation triplet auu, and is followed by the structural genes for ribosomal proteins l35 and l20 (rpmi and rplt, respectively). | 1989 | 2779520 |
| multiple amylase genes in two strains of bacillus stearothermophilus. | plasmid dna fragments from bacillus stearothermophilus atcc29609 (br135) and chromosomal dna fragments from b. stearothermophilus atcc31195 (br132) were cloned into pbr322 and transformed into escherichia coli strain hb101. clones were selected which demonstrate extracellular expression of thermostable amylase. the dna inserts from both strains showed similar restriction maps. examination of the parental strains revealed plasmids of approx. 100 kb and 35 kb for br135 and no discernible plasmids ... | 1989 | 2787266 |
| on the effects of replacing the carboxylate-binding arginine-171 by hydrophobic tyrosine or tryptophan residues in the l-lactate dehydrogenase from bacillus stearothermophilus. | for l-lactate dehydrogenases (ldh's), the interaction of the guanidinium group of their arg 171 residue with the carboxylate group of an alpha-keto acid is of primary importance in orienting the substrate productively at the active site. ldh's such as that of bacillus stearothermophilus (bsldh) are of practical importance for the preparation of chiral 2-hydroxy acids used as synthons in asymmetric synthesis but would even be more valuable in this regard if their specificities were broader. with ... | 1989 | 2790015 |
| phosphorescence properties of trp-84 and trp-310 in glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | the phosphorescence spectra of trp-84 and trp-310 in glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus in an aqueous glass show distinct 0,0 vibrational bands with peaks at 406.5 and 410.5 nm. with the aid of external heavy-atom perturbation of iodide and the thermal quenching profile, it is concluded that although both chromophores are effectively buried, only one, viz., the 406.5 nm component, is embedded in a sufficiently rigid core of the protein to phosphoresce in fl ... | 1989 | 2804244 |
| subchronic toxicity studies in dogs and in utero rats fed diets containing bacillus stearothermophilus alpha-amylase from a natural or recombinant dna host. | beagle dogs and fischer 344 rats were fed diets containing 0, 36 or 72 units bacillus stearothermophilus alpha-amylase (bsa)/g food or of bacillus subtilis alpha-amylase (cbsa)/g food. the dogs (four/sex/group) received treated diets for 13 wk. for the rat studies, the parental (f0) generation (12 males and 24 females/group for the bsa study, and 26 rats/sex/group for the cbsa study) received treated diets for 13 or 4 wk, respectively, before breeding and through weaning of the f1 pups; 20 f1 ra ... | 1989 | 2807104 |
| novel procedures for derivatization of ribosomes for crystallographic studies. | undecagold and tetrairidium clusters have been used for the preparation of heavy-metal derivatives of ribosomal particles, necessary for the evaluation of phases in the x-ray structure determination of these large particles. to obtain specific binding, monofunctional reagents of the clusters were prepared and were covalently bound to free sulfhydryl groups on the surface of the ribosome. in addition, a mutant of bacillus stearothermophilus which lacks one ribosomal protein (bl11) was grown. the ... | 1989 | 2808418 |
| double-stranded dna binding protein hu. | | 1989 | 2818428 |
| 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 |
| isolation and identification of restriction endonuclease bstf i. | | 1987 | 2821503 |
| 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 |
| 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 |
| comparative study of energy-transducing properties of cytoplasmic membranes from mesophilic and thermophilic bacillus species. | the properties of enzymes involved in energy transduction from a mesophilic (bacillus subtilis) and a thermophilic (b. stearothermophilus) bacterium were compared. membrane preparations of the two organisms contained dehydrogenases for nadh, succinate, l-alpha-glycerophosphate, and l-lactate. maximum nadh and cytochrome c oxidation rates were obtained at the respective growth temperatures of the two bacteria. the enzymes involved in the oxidation reactions in membranes of the thermophilic specie ... | 1988 | 2834342 |
| thermostable alanine racemase from bacillus stearothermophilus: dna and protein sequence determination and secondary structure prediction. | the nucleotide sequence of the alanine racemase (ec 5.1.1.1) gene from a thermophile, bacillus stearothermophilus, was determined by the dideoxy chain termination method with universal and synthetic site-specific primers. the amino acid sequence of the enzyme predicted from the nucleotide sequence was confirmed by peptide sequence information derived from the n-terminal amino acid residues and several tryptic fragments. the alanine racemase gene consists of 1158 base pairs encoding a protein of ... | 1988 | 2835089 |
| bsri, a unique restriction endonuclease from bacillus stearothermophilus which recognizes 5' actgg3'. | | 1988 | 2838810 |
| isolation and characterization of restriction endonuclease bstyi from bacillus stearothermophilus y406. | bstyi, an isoschizomer of xhoii and mfli, has been purified from bacillus stearothermophilus y406. this enzyme recognized 5'...pu/gatcpy...3' in dna and cleaved between pu and g in this sequence. bstyi can be easily isolated and purified by heparin-agarose column chromatography in a high yield (8000 units bstyi can be obtained per g wet wt of cells). | 1988 | 2839359 |
| transduction in bacillus stearothermophilus. | temperate and virulent bacteriophages isolated from soil were shown to carry out generalized transduction of bacillus stearothermophilus nub36. a transducing frequency of 1 x 10(-5) to 7 x 10(-4) was obtained for temperate phages tp-42 and tp-56. the transducing frequency for virulent phage tp-68 was two to three orders of magnitude lower. cotransfer analysis with the three phages showed that hom-1 is linked to thr-1 and that gly-1 is linked to his-1. | 1988 | 2841302 |
| conservation of short patches of amino acid sequence amongst proteins with a common function but evolutionarily distinct origins: implications for cloning genes and for structure-function analysis. | small patches of identical amino acid sequences commonly occur in proteins that have the same function but are derived from evolutionarily distant organisms. reverse translation of such patches into degenerate pools of oligonucleotides provide useful hybridization probes for cloning the gene for the corresponding protein from other organisms. since the conserved patches of identical amino acid sequence are probably important for the protein's biological function, they are preferred targets for r ... | 1988 | 2845364 |
| [a new type of cleavage of the recognition site by the site-specific endonuclease bst 4.4i from bacillus stearothermophilus 4.4]. | a site-specific endonuclease bst 4.4i was isolated from the cell extract of bacillus stearothermophilus 4.4 and partially purified by chromatography on ultragel aca-44 and heparin-sepharose. it was shown that the endonuclease cleaves lambda and m13 dna yielding distinct fragments just as endonucleases of ii and iii types but, in contrast to them can produce two two-strand cuts separated with 30 to 32 nucleotides in the region of the recognition site. | 1988 | 2847759 |
| cloning and characterization of the gene for a methanol-utilising alcohol dehydrogenase from bacillus stearothermophilus. | the cloning and characterization of the alcohol dehydrogenase (adh) gene (adh) from bacillus stearothermophilus strain dsm2334, an obligate aerobe, are described. the clone directed the synthesis of adh as judged on western blots, activity gels and tetrazolium plates. it specified an enzyme that oxidised methanol as well as ethanol. the enzyme was found to be encoded by a single gene in b. stearothermophilus which did not cross-hybridize to adh clones from escherichia coli, yeast or maize. the c ... | 1988 | 2851486 |
| the activity of glutaraldehyde against clostridium difficile. | the sporicidal activity of 2% glutaraldehyde against bacillus subtilis var. globigii and bacillus stearothermophilus was compared with that against clostridium difficile. the aerobic species, normally chosen for test purposes, survived for 2 h but cl. difficile was killed in under 10 min. in view of the impractical and lengthy immersions required to satisfy standard tests using non-pathogenic spores, a less stringent standard would probably be more appropriate. | 1985 | 2859321 |
| thermostabilization of bacillus caldolyticus glutamine synthetase by intrinsic and extrinsic factors. | | 1985 | 2866936 |
| permeability studies on protective gloves used in dental practice. | the recent guidelines issued by the dhss and the bds recommend that to prevent cross-infection gloves should be worn by dentists and their assistants whilst treating all patients. little information is available on the permeability of the various types of glove available; this pilot study was therefore conducted, to test five commonly used brands of gloves for permeability to air, electrolytes, dye and bacteria. tests were carried out before and after using the gloves for varying periods. from t ... | 1989 | 2912478 |
| pattern of action of bacillus stearothermophilus neopullulanase on pullulan. | the action of neopullulanase from bacillus stearothermophilus on many oligosaccharides was tested. the enzyme hydrolyzed not only alpha-(1----4)-glucosidic linkages but also specific alpha-(1----6)-glucosidic linkages of several branched oligosaccharides. when pullulan was used as a substrate, panose, maltose, and glucose, in that order, were produced as final products at a final molar ratio of 3:1:1. according to these results, we proposed a model for the pattern of action of neopullulanase on ... | 1989 | 2914851 |
| fructose 6-phosphate prevents the proteolyzed derivative of escherichia coli phosphofructokinase from dissociation and inactivation. | n-terminal sequence analysis shows that the limited proteolysis of escherichia coli phosphofructokinase results in the removal of the 40-50 c-terminal residues of each chain. when tetrameric, this proteolyzed derivative is still active albeit insensitive to allosteric effectors (le bras, g., and garel, j.-r. (1982) biochemistry 21, 6656-6660). in the absence of fructose 6-phosphate, the proteolyzed phosphofructokinase spontaneously loses its activity and dissociates into dimeric species. this in ... | 1985 | 2932439 |
| 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 |
| the rabbit muscle phosphofructokinase gene. implications for protein structure, function, and tissue specificity. | sequence homologies between bacterial and rabbit muscle phosphofructokinases and between the amino- and carboxyl-terminal halves of the latter suggest that the mammalian enzyme evolved from a prokaryotic progenitor by gene duplication and divergence (poorman, r. a., randolph, a., kemp, r. g., and heinrikson, r. l. (1984) nature 309, 467-469). we have isolated the gene for the rabbit enzyme and determined the nucleotide sequence for all the exons and most of the introns. this represents the first ... | 1987 | 2951385 |
| 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 |
| helping rna catalysis along. | | 1987 | 2960900 |
| 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 |
| an enzymatic method for the alpha-amylase assay which comprises a new procedure for eliminating glucose and maltose in biological fluids. | an alpha-amylase assay in biological fluids, characterized by a new procedure for eliminating glucose and/or maltose, was developed. the reagent includes thermostable glucokinase, glucosephosphate isomerase and phosphofructokinase obtained from a thermophile bacillus stearothermophilus. up to 4,000 mg/dl glucose or 600 mg/dl maltose had no effect on the measured value of alpha-amylase activity when measured at 37 degrees c, even at a serum volume fraction in the reagent of 1/50. alpha-amylase ac ... | 1988 | 2967134 |
| site-directed mutagenesis at the regulatory site of fructose 6-phosphate-1-kinase from bacillus stearothermophilus. | we have mutated arg-25, asp-59 and arg-211 to alanine; and asp-59 also to methionine, in fructose 6-phosphate-1-kinase from b. stearothermophilus (designated as ra25, da59, ra211 and dm59 respectively). all four mutants did not change the affinity of the enzyme for atp. ra25 has half the affinity for fructose 6-phosphate and exhibits sigmoidicity with respect to this substrate (hill # = 2.0). da59 has the same affinity for phosphoenolpyruvate (pep) as the wild type whereas dm59 has 3-fold the af ... | 1988 | 2972287 |
| crystallization of and preliminary crystallographic data for bacillus stearothermophilus cyclodextrin glucanotransferase. | cyclodextrin glucanotransferase from bacillus stearothermophilus tc-91 has been crystallized from an ammonium sulfate solution by the dialysis equilibrium method. the crystals belong to the orthorhombic system, space group p2(1)2(1)2(1), with cell dimensions of a = 125.5 a, b = 88.1 a, and c = 81.5 a. the crystals appear to be suitable for x-ray structure analysis, diffracting to at least 2.1 a and being resistant to radiation damage. | 1988 | 2975653 |
| crystal structure of the complex of phosphofructokinase from escherichia coli with its reaction products. | the crystal structure of escherichia coli phosphofructokinase complexed with its reaction products fructose 1,6-bisphosphate (fru1,6p) and adp/mg2+, and the allosteric activator adp/mg2+, has been determined at 2.4 a resolution. the structure was solved by molecular replacement using the known structure of bacillus stearothermophilus phosphofructokinase, and has been refined to a crystallographic r-factor of 0.165 for all data. the crystallization mixture contained the substrate fructose 6-phosp ... | 1988 | 2975709 |
| isolation of a bacillus stearothermophilus mutant exhibiting increased thermostability in its restriction endonuclease. | a procedure was developed for the selection of spontaneous mutants of bacillus stearothermophilus nub31 that are more efficient than the wild type in the restriction of phage at elevated temperatures. inactivation studies revealed that two mutants contained a more thermostable restriction enzyme and one mutant contained three times more enzyme than the wild type. the restriction endonucleases from the wild type and one of the mutants were purified to apparent homogeneity. the mutant enzyme was m ... | 1985 | 2985543 |
| replacement of mn(iii) with cu(ii) in bacillus stearothermophilus superoxide dismutase. similarity of the active site to the zinc site of copper/zinc superoxide dismutase. | copper(ii) was substituted for manganese(iii) in bacillus stearothermophilus mn-superoxide dismutase. the (epr) spectrum of the cu(ii) is distinctly rhombic, overlapping that of cu(ii) replacing zinc in copper/zinc superoxide dismutase. the copper-cyanide complex of the bacterial enzyme gives an epr spectrum nearly identical to the cyanide-copper complex of the cu/zn enzyme, indicating three nitrogens as metal ligands. these results support the suggestion that metal coordination in b. stearother ... | 1985 | 2991020 |
| partial purification and characterization of type i dna topoisomerase from bacillus stearothermophilus. | type i dna topoisomerase was partially purified from bacillus stearothermophilus by ammonium sulfate precipitation and column chromatographies on phosphocellulose, deae-cellulose and heparin-agarose. on heparin-agarose chromatography, topoisomerase i activity was separated into three fractions (designated fractions a, b, and c). each fraction was further subjected to gel filtration on sephacryl s-200. from electrophoretic analysis on polyacrylamide gel, fraction a was found to contain two enzyme ... | 1985 | 2991208 |
| isolation and partial characterization of bstvi, a thermostable isoschizomer of xhoi. | a type ii restriction endonuclease, which has been named bstvi, was isolated and partially purified from a spore-forming, gram-positive thermophilic bacilli. on the basis of its digestion patterns on various dna's, it was concluded that this enzyme is an isoschizomer of xhoi, isolated originally from xanthomonas holcicola. besides being highly thermostable, the enzyme is produced in very large amounts by this bacterial strain. a single purification step renders it free of unspecific nucleases an ... | 1985 | 2992515 |
| genetic relationship between pub110 and antibiotic-resistant plasmids obtained from thermophilic bacilli. | the molecular relationship between pub110 (kmr, 4.4 kilobases (kb] and antibiotic-resistant plasmids from thermophilic bacilli, ptht15 (tcr, 4.5 kb) and pthn1 (kmr, 4.8 kb), were studied by blot hybridization. extensive homology was observed between pub110 and ptht15 at the region which includes the replication origin. incompatibility studies revealed that ptht15 and pub110 were slightly incompatible in bacillus subtilis but that they were apparently compatible in b. stearothermophilus. this dif ... | 1985 | 2992731 |
| nucleotide sequence and promoter region for the neutral protease gene from bacillus stearothermophilus. | the thermostable neutral protease gene nprt of bacillus stearothermophilus was sequenced. the dna sequence revealed only one large open reading frame, composed of 1,644 bases and 548 amino acid residues. a shine-dalgarno sequence was found 9 bases upstream from the translation start site (atg), and the deduced amino acid sequence contained a signal sequence in its amino-terminal region. the sequence of the first 14 amino acids of purified extracellular protease completely matched that deduced fr ... | 1985 | 2993245 |
| efficient synthesis and secretion of a thermophilic alpha-amylase by protein-producing bacillus brevis 47 carrying the bacillus stearothermophilus amylase gene. | bacillus subtilis and bacillus brevis 47-5, carrying the bacillus stearothermophilus alpha-amylase gene on pub110 (pbam101), synthesized the same alpha-amylase as the donor strain as determined by the enzyme's thermal stability and nh2-terminal amino acid sequence. regardless of the host, the 34-amino acid signal peptide of the enzyme was processed at exactly the same site between two alanine residues. b. brevis 47-5(pbam101) secreted the enzyme most efficiently of the hosts examined, 100, 15, a ... | 1985 | 2999073 |
| the specific modification of histidyl residues of inorganic pyrophosphatase from bacillus stearothermophilus by photooxidation. | photooxidation of inorganic pyrophosphatase [pyrophosphate phosphohydrolase ec 3.6.1.1] from bacillus stearothermophilus in the presence of rose bengal resulted in rapid loss of enzymatic activity. the ph profile of the inactivation rate by the photooxidation showed an inflection point around ph 6.8, suggesting the involvement of histidyl residues in the inactivation. amino acid analysis revealed that the loss of enzymatic activity was accompanied by the destruction of 3 histidyl residues per mo ... | 1985 | 2999092 |
| screening for thermostable mutant of kanamycin nucleotidyltransferase by the use of a transformation system for a thermophile, bacillus stearothermophilus. | a structural gene of kanamycin nucleotidyltransferase cloned into a single-stranded bacteriophage m13 was subjected to mutagenesis with hydroxylamine. having recloned the mutagenized gene of the enzyme in a vector plasmid ptb922, the recombinant plasmid was used to transform bacillus stearothermophilus with a purpose of screening for the more thermostable enzyme than the wild type. out of greater than 8 x 10(3) transformants, 12 clones that were suspected to harbor the mutant gene encoding the m ... | 1985 | 2999110 |
| fine structure-activity analysis of mutations at position 51 of tyrosyl-trna synthetase. | residue thr-51 at the active site of tyrosyl-trna synthetase (bacillus stearothermophilus) has been replaced with all the smaller amino acids by protein engineering to investigate direct and indirect effects of mutation on substrate binding and catalysis. the gamma-hydroxyl group of thr-51 was thought to be 0.5 a too far from the ribose ring oxygen of atp to form a hydrogen bond. consistent with this, it is found that mutation of thr-51----cys-51, which should place the gamma-thiol group within ... | 1985 | 3002425 |
| 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 |
| 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 |
| 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 |
| identification of highly reactive cysteinyl and methionyl residues of rabbit muscle phosphofructokinase. | the reactivity of the 16 thiol groups of rabbit skeletal muscle phosphofructokinase has been studied extensively over the past 20 years. several of these thiols show high reactivity with a variety of reagents, display differential reactivity in the presence of allosteric ligands and substrates, and appear to be important to function because their modification changes activity and regulatory properties. in the present study, the location in the primary structure of several highly reactive thiol g ... | 1987 | 3038892 |
| structural comparison of the prokaryotic ribosomal proteins l7/l12 and l30. | the structures of two prokaryotic ribosomal proteins, the carboxyterminal half of l7/l12 from escherichia coli (l12ctf) and l30 from bacilus stearothermophilus display a remarkably similar fold in which alpha-helices pack onto one side of an antiparallel, three-stranded, beta-pleated sheet. a detailed comparison of the structures by least-squares methods reveals that more than two-thirds of the alpha carbons can be superimposed with a root mean square distance of 2.33 a. the principal difference ... | 1988 | 3047743 |
| tyrosyl-trna synthetase from baker's yeast. order of substrate addition, discrimination of 20 amino acids in aminoacylation of trnatyr-c-c-a and trnatyr-c-c-a(3'nh2). | the order of substrate addition to tyrosyl-trna synthetase from baker's yeast was investigated by bisubstrate kinetics, product inhibition and inhibition by dead-end inhibitors. the kinetic patterns are consistent with a random bi-uni uni-bi ping-pong mechanism. substrate specificity with regard to atp analogs shows that the hydroxyl groups of the ribose moiety and the amino group in position 6 of the base are essential for recognition of atp as substrate. specificity with regard to amino acids ... | 1988 | 3056726 |
| identification, cloning and sequence of the streptococcus faecium infb (translational initiation factor if2) gene. | the structural gene for translational initiation factor if2 (infb) from streptococcus faecium was identified by cross-hybridization with dna probes derived from the corresponding gene of bacillus stearothermophilus. the entire infb gene (ca. 2.8 kb) was cloned and sequenced. the amino acid sequence deduced from the nucleotide sequence shows that s. faecium initiation factor if2 (785 amino acids, mr 86,415) displays extensive homology (ca. 69% and 53%) with the region comprising three-quarters of ... | 1988 | 3063954 |
| crystallization of a ternary complex of lactate dehydrogenase from bacillus stearothermophilus. | bacillus stearothermophilus lactate dehydrogenase was purified from an overexpressing escherichia coli cell line. the enzyme has been crystallized in several different forms. all of these crystal forms were grown in the presence of nadh, sodium oxamate and fructose 1,6-bisphosphate. three crystal forms have been characterized, an orthorhombic p2(1)2(1)2 (type iii, a = 86 a, b = 105 a, c = 136 a) and two monoclinic p21 forms (type iv, a = 85 a, b = 118 a, c = 136 a, beta = 96 degrees; type v, a = ... | 1988 | 3065514 |
| [1h-nmr study of protein l7/l12 from bacterial ribosomes]. | the 500 mhz 1h-nmr spectra of dimeric protein l12 from ribosomes shows a limited number of unusually sharp signals at room temperature. this is interpreted as evidence for substantial segmental flexibility of the region in the protein molecule. we have analysed the extent of the flexible region and also the size of the organized structures of the molecule. thus residues 37-50 were found to be highly mobile whereas the n-terminal and c-terminal region are organized into folded domains. | 1988 | 3065618 |
| gene cloning and sequence determination of leucine dehydrogenase from bacillus stearothermophilus and structural comparison with other nad(p)+-dependent dehydrogenases. | the gene for leucine dehydrogenase (ec 1.4.1.9) from bacillus stearothermophilus was cloned and expressed in escherichia coli. the selection for the cloned gene was based upon activity staining of the replica printed e. coli cells. a transformant showing high leucine dehydrogenase activity was found to carry an about 9 kilobase pair plasmid, which contained 4.6 kilobase pairs of b. stearothermophilus dna. the nucleotide sequence including the 1287 base pair coding region of the leucine dehydroge ... | 1988 | 3069133 |
| crystallographic and image reconstruction studies on ribosomal particles from bacterial sources. | | 1988 | 3071693 |
| use of site-directed mutagenesis to probe the role of cys149 in the formation of charge-transfer transition in glyceraldehyde-3-phosphate dehydrogenase. | oligonucleotide-directed mutagenesis was employed to produce mutants of the glyceraldehyde-3-phosphate dehydrogenase (gapdh) of escherichia coli and bacillus stearothermophilus. three different mutants proteins--his176----asn, cys149----ser, cys149----gly--were isolated from one or both of the enzymes. the study of the properties of these mutants has shown that cys149 is clearly responsible for the information of a charge-transfer transition, named the racker band, observed during the nad+ bindi ... | 1988 | 3075757 |