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joint refinement as a tool for thorough comparison between nmr and x-ray data and structures of hu protein.joint refinement, i.e., the simultaneous refinement of a structure against both nuclear magnetic resonance (nmr) spectroscopic and x-ray crystallographic data, was performed on the hu protein from bacillus stearothermophilus (hubst). the procedure was aimed at investigating the compatibility of the two data sets and at identifying conflicting information. wherever important differences were found, such as peptide flips in the main-chain conformation, the data were further analyzed to find the ca ...200111775740
a catalase-peroxidase from a newly isolated thermoalkaliphilic bacillus sp. with potential for the treatment of textile bleaching effluents.a new thermoalkaliphilic bacterium was isolated from a textile wastewater drain and identified as a new bacillus sp. (bacillus sf). because of its high ph stability and thermostability, a catalase-peroxidase (cp) from this strain has potential for the treatment of textile bleaching effluents. the cp from bacillus sf was purified to more than 70.3-fold homogeneity using fractionated ammonium sulfate precipitation, hydrophobic interaction, and anion-exchange and gel-filtration chromatography. the ...200111778844
[synthesis of galacto-oligosaccharides by immobilized bacillus stearothermophilus].bacillus stearothermophilus was embedded in sodium alginate, chitosan, gelatin respectively and utilized for galacto-oligosaccharides (gos) production. by comparing with each other in the enzyme recovery, optimal reaction conditions, yields of gos and mechanical strength of the carriers, the gelatin was selected as better carrier for immobilization. ph, temperature, lactose concentration, lactose conversion and mass transfer resistance of carrier significantly influenced the yield of gos. the ma ...200112549092
homology modeling of bacillus subtilis tryptophanyl-trna synthetase.tryptophanyl-trna synthetase (trprs) plays a pivotal role in protein synthesis. however, till now no stereostructural data of bacillus subtilis trprs were reported. here, by using the homology modeling using bacillus stearothermophilus trprs as a template, it is demonstrated that the synthetase has 16alpha helices and 5beta sheets. the only tryptophan trp(92) is located on the interface of subunits. also the modeling presents the ligand binding site, active site and the putative binding of trna( ...200112035063
identification by saturation mutagenesis of a single residue involved in the alpha-galactosidase agab regioselectivity.the alpha-galactosidase agab of bacillus stearothermophilus displays a major 1,6 and a minor 1,3 regioselectivity. the wild-type enzyme was subjected to directed evolution (random mutagenesis and in vitro recombination) using a double screening strategy based on the elimination of the 1,6 regioselectivity and the analysis by tlc of the transglycosylation products. one of the agab mutants (e500) exhibited a new 1,2 regioselectivity and a rather high level of transglycosylation. the corresponding ...200112084981
functional grouping of thermophilic bacillus strains using amplification profiles of the 16s-23s internal spacer region.molecular and biochemical assays were used to determine the identification of thermophilic bacilli isolated from new zealand milk powder. one hundred and forty one isolates of thermophilic bacilli were classified into six species using biochemical profiles. geobacillus stearothermophilus represented 56% of the isolates. all isolates were also analysed by randomly amplified polymorphic dna (rapd) analysis, with 45 types identified. amplification of the 16s-23s rdna internal spacer region produced ...200111876361
crystal structure of binary and ternary complexes of serine hydroxymethyltransferase from bacillus stearothermophilus: insights into the catalytic mechanism.serine hydroxymethyltransferase (shmt), a member of the alpha-class of pyridoxal phosphate-dependent enzymes, catalyzes the reversible conversion of serine to glycine and tetrahydrofolate to 5,10-methylene tetrahydrofolate. we present here the crystal structures of the native enzyme and its complexes with serine, glycine, glycine, and 5-formyl tetrahydrofolate (fthf) from bacillus stearothermophilus. the first structure of the serine-bound form of shmt allows identification of residues involved ...200211877399
the thermostability of dna-binding protein hu from mesophilic, thermophilic, and extreme thermophilic bacteria.based on primary structure comparison between four highly homologous dna-binding proteins (hus) displaying differential thermostability, we have employed in vitro site-directed mutagenesis to decipher their thermostability mechanism at the molecular level. the contribution of the 11 amino acids that differ between the thermophilic hubst from bacillus stearothermophilus (tm = 61.6 degrees c) and the mesophilic hubsu from bacillus subtilis (tm = 39.7 degrees c) was evaluated by replacing these ami ...200211878558
modulation of cyclizing activity and thermostability of cyclodextrin glucanotransferase and its application as an antistaling enzyme.cyclodextrin glucanotransferase from bacillus stearothermophilus et1 (cgtase et1) is a potential antistaling enzyme with cyclodextrin (cd)-forming activity. to reduce cyclization activity of cgtase et1, phenylalanine residues at 191 and 255 were replaced with a glycine (f191g-cgtase et1) and an isoleucine (f255i-cgtase et1), respectively. temperature optima of both mutant enzymes were lower than that of the wild-type. cyclization activities of both mutants decreased dramatically, but f255i-cgtas ...200211879012
an emission pattern of a thermophilic bacteria attached to or imbedded in porous supports.there are many problems with thermophilic bacteria contamination of milk in the dairy industry. this is, in part, a result of fouling by milk components on stainless steel surfaces, which provide good harboring facilities for these bacteria to attach, imbed and grow. the interactions between milk fouling and bacteria deposited in or on the fouling deposit therefore become important issues. there have been a number of previous studies on the biofilm development in dairy processing plants. here, a ...200211883671
in situ decontamination of medical wastes using oxidative agents: a 16-month study in a polyvalent intensive care unit.over a 16-month period from september 1997 to december 1998, a prospective study was made of an on-site treatment of medical wastes in a 10-bed intensive care unit. first, the wastes were ground and then, a high concentration of ozone in air was repeatedly injected into the ground wastes. the study analysed the practical application of the system and its microbiological efficiency. inactivation experiments were made with reference strains of staphylococcus aureus, enterococcus hirae, pseudomonas ...200211886197
reaction mechanism of alanine racemase from bacillus stearothermophilus: x-ray crystallographic studies of the enzyme bound with n-(5'-phosphopyridoxyl)alanine.the crystal structures of alanine racemase bound with reaction intermediate analogs, n-(5'-phosphopyridoxyl)-l-alanine (plp-l-ala) and n-(5'-phosphopyridoxyl)-d-alanine (plp-d-ala), were determined at 2.0-a resolution with the crystallographic r factor of 17.2 for plp-l-ala and 16.9 for plp-d-ala complexes. they were quite similar not only to each other but also to the structure of the native pyridoxal 5'-phosphate (plp)-form enzyme; root mean square deviations at calpha among the three structur ...200211886871
glycon specificity profiling of alpha-glucosidases using monodeoxy and mono-o-methyl derivatives of p-nitrophenyl alpha-d-glucopyranoside.hydrolysis of probe substrates, eight possible monodeoxy and mono-o-methyl analogs of p-nitrophenyl alpha-d-glucopyranoside (pnp alpha-d-glc), modified at the c-2, c-3, c-4, and c-6 positions, was studied as part of investigations into the glycon specificities of seven alpha-glucosidases (ec 3.2.1.20) isolated from saccharomyces cerevisiae, bacillus stearothermophilus, honeybee (two enzymes), sugar beet, flint corn, and aspergillus niger. the glucosidases from sugar beet, flint corn, and a. nige ...200211909596
posttranscriptional modifications in the a-loop of 23s rrnas from selected archaea and eubacteria.posttranscriptional modifications were mapped in helices 90-92 of 23s rrna from the following phylogenetically diverse organisms: haloarcula marismortui, sulfolobus acidocaldarius, bacillus subtilis, and bacillus stearothermophilus. helix 92 is a component of the ribosomal a-site, which contacts the aminoacyl-trna during protein synthesis, implying that posttranscriptional modifications in helices 90-92 may be important for ribosome function. rna fragments were isolated from 23s rrna by site-dir ...200211911366
bacillus stearothermophilus neopullulanase selective hydrolysis of amylose to maltose in the presence of amylopectin.the specificity of bacillus stearothermophilus trs40 neopullulanase toward amylose and amylopectin was analyzed. although this neopullulanase completely hydrolyzed amylose to produce maltose as the main product, it scarcely hydrolyzed amylopectin. the molecular mass of amylopectin was decreased by only one order of magnitude, from approximately 10(8) to 10(7) da. furthermore, this neopullulanase selectively hydrolyzed amylose when starch was used as a substrate. this phenomenon, efficient hydrol ...200211916682
potassium functionally replaces the second lysine of the kmsks signature sequence in human tyrosyl-trna synthetase.unlike their bacterial homologues, a number of eukaryotic tyrosyl-trna synthetases require potassium to catalyze the aminoacylation reaction. in addition, the second lysine in the class i-specific kmsks signature motif is absent from all known eukaryotic tyrosyl-trna synthetase sequences, except those of higher plants. this lysine, which is the most highly conserved residue in the class i aminoacyl-trna synthetase family, has been shown to interact with the pyrophosphate moiety of the atp substr ...200211927599
a recombinant bacterial cell surface (s-layer)-major birch pollen allergen-fusion protein (rsbsc/bet v1) maintains the ability to self-assemble into regularly structured monomolecular lattices and the functionality of the allergen.the mature crystalline bacterial cell surface (s-layer) protein sbsc of bacillus stearothermophilus atcc 12980 comprises amino acids 31-1099 and assembles into an oblique lattice type. as the deletion of up to 179 c-terminal amino acids did not interfere with the self-assembly properties of sbsc, the sequence encoding the major birch pollen allergen (bet v1) was fused to the sequence encoding the truncated form rsbsc(31-920). the s-layer fusion protein, termed rsbsc/bet v1, maintained the abilit ...200211932495
the identification of the acid-base catalyst of alpha-arabinofuranosidase from geobacillus stearothermophilus t-6, a family 51 glycoside hydrolase.the alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6 (abfa t-6) belongs to the retaining family 51 glycoside hydrolases. the conserved glu175 was proposed to be the acid-base catalytic residue. abfa t-6 exhibits residual activity towards aryl beta-d-xylopyranosides. this phenomenon was used to examine the catalytic properties of the putative acid-base mutant e175a. data from kinetic experiments, ph profiles, azide rescue, and the identification of the xylopyranosyl azide produ ...200211943144
a novel fluorescence competitive assay for glucose determinations by using a thermostable glucokinase from the thermophilic microorganism bacillus stearothermophilus.we describe the use of a thermostable glucokinase in a novel competitive fluorescence assay for glucose. glucokinase from bacillus stearothermophilus (bsgk) was found to retain enzymatic activity in solution for over 20 days. the single cysteine residue in bsgk, which is near the active site, was labeled with a fluorescent probe, 2-(4-iodoacetamidoanilino)naphthalene-6-sulfonic acid. the ans-labeled bsgk displayed a modest 25% decrease in the emission intensity upon binding glucose but no change ...200211950213
crystal structure of the bacillus stearothermophilus anti-sigma factor spoiiab with the sporulation sigma factor sigmaf.cell type-specific transcription during bacillus sporulation is established by sigmaf. spoiiab is an anti-sigma that binds and negatively regulates sigmaf, as well as a serine kinase that phosphorylates and inactivates the anti-anti-sigma spoiiaa. the crystal structure of sigmaf bound to the spoiiab dimer in the low-affinity, adp form has been determined at 2.9 a resolution. spoiiab adopts the ghkl superfamily fold of atpases and histidine kinases. a domain of sigmaf contacts both spoiiab monome ...200211955433
thermodynamic analysis of the binding of component enzymes in the assembly of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.the peripheral subunit-binding domain (psbd) of the dihydrolipoyl acetyltransferase (e2, ec 2.3.1.12) binds tightly but mutually exclusively to dihydrolipoyl dehydrogenase (e3, ec 1.8.1.4) and pyruvate decarboxylase (e1, ec 1.2.4.1) in the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus. isothermal titration calorimetry (itc) experiments demonstrated that the enthalpies of binding (deltah degrees ) of both e3 and e1 with the psbd varied with salt concentration, temperat ...200211967366
cooperative action of alpha-glucanotransferase and maltogenic amylase for an improved process of isomaltooligosaccharide (imo) production.maltogenic amylase and alpha-glucanotransferase (alpha-gtase) were employed in an effort to develop an efficient process for the production of isomaltooligosaccharides (imos). bacillus stearothermophilus maltogenic amylase (bsma) and alpha-gtase from thermotoga maritima were overexpressed in escherichia coli using overexpression vectors. an imo mixture containing 58% of various imos was produced from liquefied corn syrup by the hydrolyzing and transglycosylation activities of bsma alone. when bs ...200211982404
modeling and kinetic analysis of the reaction system using whole cells with separately and co-expressed d-hydantoinase and n-carbamoylase.we developed a kinetic model that describes a heterogeneous reaction system for the production of d-p-hydroxyphenylglycine from d,l-p-hydroxyphenyl-hydantoin using d-hydantoinase of bacillus stearothermophilus sd1 and n-carbamoylase of agrobacterium tumefaciens nrrl b11291. as a biocatalyst, whole cells with separately or co-expressed enzymes were used. the reaction system involves dissolution of substrate particles, enzymatic conversion, racemization of the l-form substrate, and transfer of the ...200212001170
enhancement of proteolytic enzyme activity excreted from bacillus stearothermophilus for a thermophilic aerobic digestion process.proteolysis is one of the main enzymatic reactions involved in waste activated sludge (was) digestion. in this study, proteases excreted from bacillus stearothermophilus (atcc 31197) were classified, and an enhancement of protease activity was achieved using economical chemical additives for was digestion. proteases excreted from b. stearothermophilus were classified into two families: serine and metallo-proteases. various metal ions were investigated as additives which could potentially enhance ...200212003317
requirement for c-terminal extension to the rna binding domain for efficient rna binding by ribosomal protein l2.ribosomal protein l2 is a primary 23s rrna binding protein in the large ribosomal subunit. we examined the contribution of the n- and c-terminal regions of bacillus stearothermophilus l2 (bstl2) to the 23s rrna binding activity. the mutant desn, in which the n-terminal 59 residues of bstl2 were deleted, bound to the 23s rrna fragment to the same extent as wild type bstl2, but the mutation desc, in which the c-terminal 74 amino acid residues were deleted, abolished the binding activity. these obs ...200212005072
structure and dynamics of the anticodon arm binding domain of bacillus stearothermophilus tyrosyl-trna synthetase.the structure of a recombinant protein, tyrrs(delta4), corresponding to the anticodon arm binding domain of bacillus stearothermophilus tyrosyl-trna synthetase, has been solved, and its dynamics have been studied by nuclear magnetic resonance (nmr). it is the first structure described for such a domain of a tyrosyl-trna synthetase. it consists of a five-stranded beta sheet, packed against two alpha helices on one side and one alpha helix on the other side. a large part of the domain is structura ...200212005430
modification of ascorbic acid using transglycosylation activity of bacillus stearothermophilus maltogenic amylase to enhance its oxidative stability.ascorbic acid (1), a natural antioxidant, was modified by employing transglycosylation activity of bacillus stearothermophilus maltogenic amylase with maltotriose and acarbose as donor molecules to enhance its oxidative stability. the transglycosylation reaction with maltotriose as donor created mono- and di-glycosyl transfer products with an alpha-(1,6)-glycosidic linkage. in addition, two acarviosine-glucosyl transfer products were generated when transglycosylation was performed with acarbose ...200212010003
isolation of chimaeric forms of elongation factor ef-tu by affinity chromatography.six different recombinant chimaeric forms of a three-domain protein, proteosynthetic elongation factor tu (ef-tu), composed of domains of ef-tu of mesophilic (escherichia coli) and thermophilic (bacillus stearothermophilus) origin as well as free n-terminal domains of ef-tu, and the whole recombinant ef-tus of both organisms were prepared and isolated by the gst (glutathione s-transferase) fusion technology. several modifications in the standard isolation and purification procedures are describe ...200212013219
comparison of the catalytic roles played by the kmsks motif in the human and bacillus stearothermophilus trosyl-trna synthetases.the class i aminoacyl-trna synthetases are characterized by two signature sequence motifs, "high" and "kmsks." in bacillus stearothermophilus tyrosyl-trna synthetase, the kmsks motif (230kfgkt234) has been shown to stabilize the transition state for tyrosine activation through interactions with the pyrophosphate moiety of atp. in most eukaryotic tyrosyl-trna synthetases, the second lysine in the kmsks motif is replaced by a serine or an alanine residue. recent kinetic studies indicate that potas ...200212016229
the effect of bioindicator preparation and storage on thermal resistance of bacillus stearothermophilus spores.paper strips inoculated with spores of bacillus stearothermophilus atcc 7953 were conventionally dried (lot 1) and lyophilized (lot 2); stored in defined environments of 32 and 86% relative humidity at 10, 25 and 33 degrees c for 210 d; and submitted to moist heat treatments at 121 degrees c. a significant decrease in thermal resistance from initial starting levels was found for lyophilized bioindicators stored at 86% relative humidity. the respective average d121 degrees c values were 1.55+/-0. ...200212018279
the effect of composition of parenteral solution on the thermal resistance of bacillus stearothermophilus and bacillus subtilis spores.large-volume parenteral solutions were submitted to heat treatments after being inoculated with bacillus stearothermophilus atcc 7953 (tr = 121 degrees c) and bacillus subtilis atcc 9372 (tr = 104.5 degrees c) spores. the average decimal reduction time for b. stearothermophilus ranged from a d121 degrees c value of 1.31 to 3.14 min, in glucophysiologic and ringer's solutions respectively. for b. subtilis, d104.5 degrees c value increased from 0.69 to 1.37 min, in ringer's (ph=5.91) and 50% gluco ...200212018280
transition state stabilization by the n-terminal anticodon-binding domain of lysyl-trna synthetase.lysyl-trna synthetase from bacillus stearothermophilus (b.s. lysrs) (ec ) catalyzes aminoacylation of trna(lys) with l-lysine, in which l-lysine was first activated with atp to yield an enzyme (lysyladenylate complex), and then the lysine molecule was transferred from the complex to trna(lys). b.s. lysrs is a homodimeric enzyme with a subunit that consists of two domains, an n-terminal trna anticodon-binding domain (tab-nd: ser(1)-pro(144)) and a c-terminal class ii-specific catalytic domain (ca ...200212019264
thermoadaptation of alpha-galactosidase agab1 in thermus thermophilus.the evolutionary potential of a thermostable alpha-galactosidase, with regard to improved catalytic activity at high temperatures, was investigated by employing an in vivo selection system based on thermophilic bacteria. for this purpose, hybrid alpha-galactosidase genes of agaa and agab from bacillus stearothermophilus kve39, designated agaa1 and agab1, were cloned into an autonomously replicating thermus vector and introduced into thermus thermophilus of1053gd (deltaagat) by transformation. th ...200212029056
overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from bacillus stearothermophilus.serine hydroxymethyltransferase (shmt), a pyridoxal-5' -phosphate (plp) dependent enzyme catalyzes the interconversion of l-ser and gly using tetrahydrofolate as a substrate. the gene encoding for shmt was amplified by pcr from genomic dna of bacillus stearothermophilus and the pcr product was cloned and overexpressed in escherichia coli. the purified recombinant enzyme was isolated as a mixture of dimer (90%) and tetramer (10%). this is the first report demonstrating the existence of shmt as a ...200212089472
an engineered escherichia coli tyrosyl-trna synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system.tyrosyl-trna synthetase (tyrrs) from escherichia coli was engineered to preferentially recognize 3-iodo-l-tyrosine rather than l-tyrosine for the site-specific incorporation of 3-iodo-l-tyrosine into proteins in eukaryotic translation systems. the wild-type tyrrs does not recognize 3-iodo-l-tyrosine, because of the bulky iodine substitution. on the basis of the reported crystal structure of bacillus stearothermophilus tyrrs, three residues, y37, q179, and q195, in the l-tyrosine-binding site wer ...200212097643
thermal unfolding used as a probe to characterize the intra- and intersubunit stabilizing interactions in phosphorylating d-glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus.tetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus can be described as a dimer of dimers with three nonequivalent interfaces. to investigate the contribution of intra- and intersubunit interactions to gapdh thermostability, 10 residues located either at the cofactor domain (amino acids 1-148 and 313-333) or at the catalytic domain (amino acids 149-312) were mutated and the thermal unfolding of the mutants was studied by differential scann ...200212056886
microwave-assisted rapid characterization of lipase selectivities.a rapid screening procedure for characterization of lipase selectivities using microwaves was developed. the rate of reaction of various commercial lipases (porcine pancreas, mucor miehei, candida rugosa, pseudomonas cepacia) as well as lipases from laboratory isolates-bacillus stearothermophilus and burkholderia cepacia rgp-10 for triolein hydrolysis was 7- to 12-fold higher in a microwave oven as compared to that by ph stat. the esterification of sucrose/methanol and ascorbic acid with differe ...200212062110
immobilized bacterial spores for use as bioindicators in the validation of thermal sterilization processes.spores of bacillus subtilis atcc 6051 and bacillus stearothermophilus nctc 10003 were immobilized in monodisperse alginate beads (diameter, 550 microm +/- 5%), and the capacity of the immobilized bioindicators to provide accurate and reliable f-values for sterilization processes was studied. the resistance of the beads to abrasion and heat was strong enough to ensure total retention of the bioindicators in the beads in a sterilization cycle. d- and z-values for free spores were identical to thos ...200212117247
crystal structure of d-hydantoinase from bacillus stearothermophilus: insight into the stereochemistry of enantioselectivity.industrial production of antibiotics, such as semisynthetic penicillins and cephalosporins, requires optically pure d-p-hydroxylphenylglycine and its derivatives as important side-chain precursors. to produce optically pure d-amino acids, microbial d-hydantoinase (e.c. 3.5.2.2) is used for stereospecific hydrolysis of chemically synthesized cyclic hydantoins. we report the apo-crystal structure of d-hydantoinase from b. stearothermophilus sd1 at 3.0 a resolution. the structure has a classic tim ...200212135362
improved partitioning in aqueous two-phase system of tyrosine-tagged recombinant lactate dehydrogenase.the partitioning of bacillus stearothermophilus lactate dehydrogenase (ldh) in an aqueous two-phase system was studied. particularly, the influence of tyrosine tags on the partitioning was evaluated. the hydrophobic effect, caused by the addition of tyrosine residues, was determined in a system based on dextran and the thermoseparating ethylene oxide-propylene oxide random copolymer (eo30po70). five different ldh variants were constructed with n-terminal tags containing tyrosines (y3 and y6), ty ...200212135559
boilysin and thermolysin in dipeptide synthesis: a comparative study.boilysin (bln) is an engineered, highly thermostable neutral protease from bacillus stearothermophilus. its high resistance is based on the stabilization of a surface loop (amino acid residues 55-69) by eight amino acid exchanges, including the introduction of a disulphide bond. in the present study, bln was compared with the well-known and structurally related thermolysin (from b. thermoproteolyticus) with respect to their dipeptide- synthetic properties. the synthesis of the aspartame precurso ...200212149125
identification of key amino acid residues in the assembly of enzymes into the pyruvate dehydrogenase complex of bacillus stearothermophilus: a kinetic and thermodynamic analysis.structural studies have shown that electrostatic interactions play a major part in the binding of dihydrolipoyl dehydrogenase (e3) to the peripheral subunit-binding domain (psbd) of the dihydrolipoyl acyltransferase (e2) in the assembly of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus. the binding is characterized by a small, unfavorable enthalpy change (deltah degrees = +2.2 kcal/mol) and a large, positive entropy change (tdeltas degrees = +14.8 kcal/mol). the co ...200212173931
expression of bacillus stearothermophilus lv cadmium resistance genes in escherichia coli causes hypersensitivity to cadmium chloride.determination of the nucleotide sequence of a 4.5-kb chromosomal dna fragment of bacillus stearothermophilus lv revealed two open reading frames (orfs) of 121 and 727 amino acids (aa) that exhibit a high degree of similarity with the cadc and cada cadmium resistance genes of a number of microorganisms. transfer and expression of the b. stearothermophilus lv cada or cadc/ cada genes in e. coli caused increased cadmium chloride susceptibility in the bacterial host. transfer of cadc alone did not r ...200212177740
aerobic metabolism of phenylacetic acids in azoarcus evansii.the aerobic metabolism of phenylacetic acid (pa) and 4-hydroxyphenylacetic acid (4-ohpa) was investigated in the beta-proteobacterium azoarcus evansii. evidence for the existence of two independent catabolic pathways for pa and 4-ohpa is presented. 4-ohpa metabolism involves the formation of 2,5-dihydroxyphenylacetate (homogentisate) and maleylacetoacetate catalyzed by specifically induced 4-ohpa 1-monooxygenase and homogentisate 1,2-dioxygenase. the metabolism of pa starts by its activation to ...200212189419
site-directed mutagenesis of tyr354 in geobacillus stearothermophilus alanine racemase identifies a role in controlling substrate specificity and a possible role in the evolution of antibiotic resistance. 200212203980
detailed kinetic analysis and identification of the nucleophile in alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6, a family 51 glycoside hydrolase.alpha-l-arabinofuranosidases cleave the l-arabinofuranoside side chains of different hemicelluloses and are key enzymes in the complete degradation of the plant cell wall. the alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6, a family 51 glycoside hydrolase, was subjected to a detailed mechanistic study. aryl-alpha-l-arabinofuranosides with various leaving groups were synthesized and used to verify the catalytic mechanism and catalytic residues of the enzyme. the steady-state ...200212221104
site-directed mutagenesis reveals roles for conserved amino acid residues in the hexameric dna helicase dnab from bacillus stearothermophilus.site-directed mutagenesis studies on conserved amino acid residues within motifs h1, h1a, h2 and h3 of the hexameric replicative helicase dnab from bacillus stearothermophilus revealed specific functions associated with these residues. in particular, residues that coordinate a bound mg2+ in the active site (t217 and d320) are important for the function of the enzyme but are not required for the formation of stable hexamers. a conserved glutamic acid (e241) in motif h1a is likely to be involved i ...200212235389
prfa protein of bacillus species: prediction and demonstration of endonuclease activity on dna.the prfa gene product of gram-positive bacteria is unusual in being implicated in several cellular processes; cell wall synthesis, chromosome segregation, and dna recombination and repair. however, no homology of prfa with other proteins has been evident. here we report a structural relationship between prfa and the restriction enzyme pvuii, and thereby produce models that predict that prfa binds dna. indeed, wild-type bacillus stearothermophilus prfa, but not a catalytic site mutant, nicked one ...200212237459
the purification and characterization of a bacillus stearothermophilus methionine aminopeptidase (metap).methionine aminopeptidase (metap) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. the enzyme was purified to homogeneity from bacillus stearothermophilus (kctc 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including deaesepharose ion exchange, hydroxylapatite, ultrogel aca 54 gel filtration, and reactive red 120 dye affinity chromatography). the apparent molecular masses of the enzyme were 81,300 da and 41,00 ...200212297034
different unfolding pathways for mesophilic and thermophilic homologues of serine hydroxymethyltransferase.to determine how much information can be transferred from folding and unfolding studies of one protein to another member of the same family or between the mesophilic and thermophilic homologues of a protein, we have characterized the equilibrium unfolding process of the dimeric enzyme serine hydroxymethyltransferase (shmt) from two sources, bacillus subtilis (bsshmt) and bacillus stearothermophilus (bstshmt). although the sequences of the two enzymes are highly identical ( approximately 77%) and ...200212356312
angiotensin i-converting enzyme inhibitory peptides derived from wakame (undaria pinnatifida) and their antihypertensive effect in spontaneously hypertensive rats.seven kinds of angiotensin i-converting enzyme (ace) inhibitory peptides were isolated from the hydrolysates of wakame (undaria pinnatifida) by protease s "amano" (from bacillus stearothermophilus) by using three-step high-performance liquid chromatography (hplc) on a reverse-phase column. these peptides were identified by amino acid composition analysis, sequence analysis, and liquid chromatography-mass spectrometry (lc-ms), as val-tyr (ic(50) = 35.2 microm), ile-tyr (6.1 microm), ala-trp (18.8 ...200212358510
inhibition of adhesion of enterotoxigenic escherichia coli k88 by soya bean tempe.tempe is a traditional fungal fermented food made from soaked and cooked soya beans. it has been associated with antidiarrhoeal characteristics. this study investigated potential inhibitory effects of tempe on enterotoxigenic escherichia coli (etec) k88.200212358694
proteomic analysis of a thermostable superoxide dismutase from bacillus stearothermophilus tls33.thermophilic bacterium bacillus stearothermophilus tls33 isolated from a hot spring in chiang mai, thailand produces an extracellular superoxide dismutase (sod). sod is a free radical metabolizing enzyme that protects the cell membrane from damage by the highly reactive superoxide free radicals. to identify the secreted sod, we used the systematically proteomic approaches of two-dimensional polyacrylamide gel electrophoresis (2-d page) analysis and database searching. the bacterium was grown in ...200212362349
proteomic study of cold shock protein in bacillus stearothermophilus p1: comparison of temperature downshifts.the thermophilic bacterium bacillus stearothermophilus p1 is unique in its ability to thrive in extreme environments such as high temperatures or high ph conditions. the study of cold shock response is very interesting and interpreted as a shock response to express the genes involved in synthesis of specific proteins. this study investigated the study of cold shock protein of b. stearothermophilus p1 when the cell culture temperature shifted from 65 degrees c to 37 degrees c and 25 degrees c. ce ...200212362350
mapping and sequencing of cardiolipins from geobacillus stearothermophilus nrs 2004/3a by positive and negative ion nanoesi-qtof-ms and ms/ms.in the course of systematic studies on surface layer (s-layer) glycoproteins of bacilli, the chloroform/methanol extract from whole cells of geobacillus stearothermophilus nrs 2004/3a has been submitted to ms analysis. glucosylated cardiolipins were found as minor components of the total lipid and phospholipid mixture by de novo identification. after purification of the crude extract using a combined column chromatography/2d tlc protocol, structural investigations of components in the lipid frac ...200212375283
trosy-nmr studies of the 91kda trap protein reveal allosteric control of a gene regulatory protein by ligand-altered flexibility.the tryptophan biosynthesis genes of several bacilli are controlled through terminator/anti-terminator transcriptional attenuation. this process is regulated by tryptophan-dependent binding of the trp rna-binding attenuation protein (trap) to the leader region of the trp operon mrna, precluding formation of the antiterminator rna hairpin, and allowing formation of the less stable terminator hairpin. crystal structures are available of trap in complex with tryptophan and in ternary complex with t ...200212381302
reversible ligand-induced dissociation of a tryptophan-shift mutant of phosphofructokinase from bacillus stearothermophilus.the biophysical properties of a tryptophan-shifted mutant of phosphofructokinase from bacillus stearothermophilus (bspfk) have been examined. the mutant, designated w179y/y164w, has kinetic and thermodynamic properties similar to the wild-type enzyme. a 2-fold decrease in kcat is observed, and the mutant displays a 3-fold smaller k(0.5) for the substrate, fructose-6-phosphate (fru-6-p), as compared to the wild-type enzyme. the dissociation constant for the inhibitor, phospho(enol)pyruvate (pep), ...200212390023
genes coding for a new pathway of aerobic benzoate metabolism in azoarcus evansii.a new pathway for aerobic benzoate oxidation has been postulated for azoarcus evansii and for a bacillus stearothermophilus-like strain. benzoate is first transformed into benzoyl coenzyme a (benzoyl-coa), which subsequently is oxidized to 3-hydroxyadipyl-coa and then to 3-ketoadipyl-coa; all intermediates are coa thioesters. the genes coding for this benzoate-induced pathway were investigated in the beta-proteobacterium a. evansii. they were identified on the basis of n-terminal amino acid sequ ...200212399500
characterization of trehalose phosphorylase from bacillus stearothermophilus sk-1 and nucleotide sequence of the corresponding gene.a bacterial trehalose phosphorylase (tpase; ec 2.4.1.64) was purified from the culture supernatant of bacillus stearothermophilus sk-1 to apparent homogeneity, and some properties were investigated. furthermore, a gene from sk-1 responsible for the tpase was cloned by southern hybridization with a degenerate oligonucleotide probe synthesized on the basis of the n-terminal sequence of the purified enzyme. the mr of the enzyme was estimated to be 150,000 by gel filtration and 83,000 by sds-page, s ...200212400680
influence of induced fit in the interaction of bacillus subtilis trp rna-binding attenuator protein and its rna antiterminator target oligomer.in the presence of excess tryptophan, tryptophan-activated trap (trp rna-binding attenuator protein) binds to a specific target in the trp-leader transcript, which induces the formation of a transcription terminator and transcription halts in the leader region. in the absence of tryptophan, trap does not bind rna, an antiterminator forms, and the operon is expressed. although the ternary complex involving trap (bacillus stearothermophilus), tryptophan, and the rna target has recently been crysta ...200212402353
evolution of signalling in the sporulation phosphorelay.two-component and phosphorelay signal transduction systems are believed to function as environ-mental sensors that programme gene expression to the composition of the ecological niche in which a microbe normally resides. the question of how evolutionarily related bacteria that occupy different environments change their signal transduction pathways to adapt to such environments was asked of the sporulation phosphorelay of bacillus subtilis, bacillus halodurans, bacillus anthracis and bacillus ste ...200212406209
site-specific incorporation of an unnatural amino acid into proteins in mammalian cells.a suppressor trna(tyr) and mutant tyrosyl-trna synthetase (tyrrs) pair was developed to incorporate 3-iodo-l-tyrosine into proteins in mammalian cells. first, the escherichia coli suppressor trna(tyr) gene was mutated, at three positions in the d arm, to generate the internal promoter for expression. however, this trna, together with the cognate tyrrs, failed to exhibit suppressor activity in mammalian cells. then, we found that amber suppression can occur with the heterologous pair of e.coli ty ...200212409460
crystal structure of a thermostable lipase from bacillus stearothermophilus p1.we describe the first lipase structure from a thermophilic organism. it shares less than 20% amino acid sequence identity with other lipases for which there are crystal structures, and shows significant insertions compared with the typical alpha/beta hydrolase canonical fold. the structure contains a zinc-binding site which is unique among all lipases with known structures, and which may play a role in enhancing thermal stability. zinc binding is mediated by two histidine and two aspartic acid r ...200212417199
arginine operator binding by heterologous and chimeric argr repressors from escherichia coli and bacillus stearothermophilus.bacillus stearothermophilus argr binds efficiently to the escherichia coli carab operator, whereas the e. coli repressor binds very poorly to the argco operator of b. stearothermophilus. in order to elucidate this contradictory behavior between argrs, we constructed chimeric proteins by swapping n-terminal dna-binding and c-terminal oligomerization domains or by exchanging the linker peptide. chimeras carrying the e. coli dna-binding domain and the b. stearothermophilus oligomerization domain sh ...200212426349
isolation of glucocardiolipins from geobacillus stearothermophilus nrs 2004/3a.glucose-substituted cardiolipins account for about 4 mol% of total phospholipid extracted from exponentially grown cells of geobacillus stearothermophilus nrs 2004/3a. individual glucocardiolipin species exhibited differences in fatty acid substitution, with iso-c(15:0) and anteiso-c(17:0) prevailing. the compounds were purified to homogeneity by a novel protocol and precharacterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.200212426359
the 2b domain of the escherichia coli rep protein is not required for dna helicase activity.the escherichia coli rep protein is a 3' to 5' sf1 dna helicase required for replication of bacteriophage phix174 in e. coli, and is structurally homologous to the e. coli uvrd helicase and the bacillus stearothermophilus pcra helicase. previous crystallographic studies of rep protein bound to single-stranded dna revealed that it can undergo a large conformational change consisting of an approximately 130 degrees rotation of its 2b subdomain about a hinge region connected to the 2a subdomain. ba ...200212441398
stable ammonia-specific nad synthetase from bacillus stearothermophilus: purification, characterization, gene cloning, and applications.bacillus stearothermophilus h-804 isolated from a hot spring in beppu, japan, produced an ammonia-specific nad synthetase (ec 6.3.1.5). the enzyme specifically used nh3 as an amide donor for the synthesis of nad as it formed amp and pyrophosphate from deamide-nad and atp. none of the l-amino acids tested, such as l-asparagine or l-glutamine, or other amino compounds such as urea, uric acid, or creatinine was used instead of nh3. mg2+ was needed for the activity, and the maximum enzyme activity w ...200212450114
studies on bacillus stearothermophilus. part ii. transformation of progesterone.bacillus stearothermophilus, a thermophilic bacterium isolated from kuwaiti desert, when incubated with exogenous progesterone for 10 days at 65 degrees c produced two new dihydroxy isomers of progesterone, and two known compounds, 5 alpha-pregnane-3,6,20-trione and 6-dehydroprogesterone, along with the earlier reported monohydroxylated metabolites and a b-seco compound. the two new dihydroxy compounds were identified as 6 alpha,20 alpha-dihydroxyprogesterone and 6 beta,20 alpha-dihydroxyprogest ...200212477492
[analysis of function of the 452 bp sequence proceeding the pgib gene in bacillus stearothermophilus].by random mutagenesis with chemical mutagen n-methyl-n'-nitro-n-nitrosoguanidine, a shuttle promoter probe vetor mutant ppgv5 (tr65) was obtained. a single site mutation from g to t was discovered at nucleotide + 238 of kan gene encoding kanamycin nucleotidyltransferae by sequence analysis. using the gene pheb that encodes thermostable catechol 2'3-dioxygenase from bacillus stearothermophilus fdtp-3 as the reporter gene, a transcriptional fusion plasmid ppgvpb452 was constructed and electroporta ...200212557393
detailed analysis of rna-protein interactions within the bacterial ribosomal protein l5/5s rrna complex.the crystal structure of ribosomal protein l5 from thermus thermophilus complexed with a 34-nt fragment comprising helix iii and loop c of escherichia coli 5s rrna has been determined at 2.5 a resolution. the protein specifically interacts with the bulged nucleotides at the top of loop c of 5s rrna. the rrna and protein contact surfaces are strongly stabilized by intramolecular interactions. charged and polar atoms forming the network of conserved intermolecular hydrogen bonds are located in two ...200212515387
expression, purification, and crystallization of glutamyl-trna(gln) specific amidotransferase from bacillus stearothermophilus.although the genes that encode the glutamyl-trna(gln) (glu-trna(gln)) specific amidotransferase (glu-adtase) from various bacteria and eukaryotic organelles are known, the precise mechanism of the enzyme is still unclear. one of the reasons is that there is no information on the three-dimensional structure of the complex, the glu-adtase:glu-trna(gln):atp:amino group donor. to obtain the crystals of glu-adtase, the glu-adtase of bacillus stearothermophilus was overexpressed and purified after clo ...200212521300
crystal structures of the bacillus stearothermophilus cca-adding enzyme and its complexes with atp or ctp.cca-adding enzymes polymerize cca onto the 3' terminus of immature trnas without using a nucleic acid template. the 3.0 a resolution crystal structures of the cca-adding enzyme from bacillus stearothermophilus and its complexes with atp or ctp reveal a seahorse-shaped subunit consisting of four domains: head, neck, body, and tail. the head is structurally homologous to the palm domain of dna polymerase beta but has additional structural features and functions. the neck, body, and tail represent ...200212526808
cloning of the maltose phosphorylase gene from bacillus sp. strain rk-1 and efficient production of the cloned gene and the trehalose phosphorylase gene from bacillus stearothermophilus sk-1 in bacillus subtilis.the maltose phosphorylase (mpase) gene of bacillus sp. strain rk-1 was cloned by pcr with oligonucleotide primers designed on the basis of a partial n-terminal amino acid sequence of the purified enzyme. the mpase gene consisted of 2,655 bp encoding a theoretical protein with a mr of 88,460, and had no secretion signal sequence, although most of the mpase activity was detected in the culture supernatant of rk-1. this cloned mpase gene and the trehalose phosphorylase (tpase) gene from bacillus st ...200212596853
a structure-based design approach for the identification of novel inhibitors: application to an alanine racemase.we report a new structure-based strategy for the identification of novel inhibitors. this approach has been applied to bacillus stearothermophilus alanine racemase (alar), an enzyme implicated in the biosynthesis of the bacterial cell wall. the enzyme catalyzes the racemization of l- and d-alanine using pyridoxal 5'-phosphate (plp) as a cofactor. the restriction of alar to bacteria and some fungi and the absolute requirement for d-alanine in peptidoglycan biosynthesis make alanine racemase a sui ...200212825624
compact form of dna induced by dna-binding protein hu.interaction of dna-binding protein hu from bacillus stearothermophilus (hubst) with coliphage t2 dna was investigated by means of a single-duplex dna chain visualization method using fluorescence microscopy. fluorescence microscopic images of coliphage t2 dna molecules were observed as a function of hubst concentration. the average fluorescence image size of t2 dna decreased with increase in hubst concentration to a size comparable to that of a dna globule induced by polyethylene glycol (peg) an ...200211779206
transcription regulation in thermophilic bacteria: high resolution contact probing of bacillus stearothermophilus and thermotoga neapolitana arginine repressor-operator interactions.arginine-mediated regulation is remarkably well conserved in very divergent bacteria, and shows a number of unusual features that distinguish arginine regulation from other transcriptional control mechanisms. the arginine repressor subunit consists of a basic n-terminal dna-binding domain, which belongs to the winged helix-turn-helix family, connected through a flexible linker to an acidic c-terminal domain responsible for binding of arginine and assembly of the high-affinity holohexamer, which ...200211786010
the anti-trp rna-binding attenuation protein (anti-trap), at, recognizes the tryptophan-activated rna binding domain of the trap regulatory protein.in bacillus subtilis, the trp rna-binding attenuation protein (trap) regulates expression of genes involved in tryptophan metabolism in response to the accumulation of l-tryptophan. tryptophan-activated trap negatively regulates expression by binding to specific mrna sequences and either promoting transcription termination or blocking translation initiation. conversely, the accumulation of uncharged trna(trp) induces synthesis of an anti-trap protein (at), which forms a complex with trap and inh ...200211786553
the surface layer (s-layer) glycoprotein of geobacillus stearothermophilus nrs 2004/3a. analysis of its glycosylation.geobacillus stearothermophilus nrs 2004/3a possesses an oblique surface layer (s-layer) composed of glycoprotein subunits as the outermost component of its cell wall. in addition to the elucidation of the complete s-layer glycan primary structure and the determination of the glycosylation sites, the structural gene sgse encoding the s-layer protein was isolated by polymerase chain reaction-based techniques. the open reading frame codes for a protein of 903 amino acids, including a leader sequenc ...200211741945
expression, purification, crystallization and preliminary crystallographic analysis of a thermostable lipase from bacillus stearothermophilus p1.the gene encoding a thermostable lipase secreted by bacillus stearothermophilus p1 has been cloned and overexpressed in escherichia coli. the recombinant lipase was purified to homogeneity using ammonium sulfate precipitation, anion-exchange chromatography (poros 20 hq) and sephacryl s-200hr. the molecular mass was shown to be 43 209 da by mass spectrometry. crystals suitable for x-ray diffraction analysis were obtained by the hanging-drop method of vapour diffusion with ammonium sulfate as the ...200211752807
identification and properties of type i-signal peptidases of bacillus amyloliquefaciens.the use of bacillus amyloliquefaciens for enzyme production and its exceptional high protein export capacity initiated this study where the presence and function of multiple type i signal peptidase isoforms was investigated. in addition to type i signal peptidases sips(ba) [meijer, w.j.j., de jong, a., bea, g., wisman, a., tjalsma, h., venema, g., bron, s. & van dijl, j.m. (1995) mol. microbiol. 17, 621-631] and sipt(ba) [hoang, v. & hofemeister, j. (1995) biochim. biophys. acta 1269, 64-68] whi ...200211856304
catalysis of tyrosyl-adenylate formation by the human tyrosyl-trna synthetase.although the active site residues in the bacillus stearothermophilus and human tyrosyl-trna synthetases are largely conserved, several differences exist between the two enzymes. in particular, three amino acids that stabilize the transition state for the activation of tyrosine in b. stearothermophilus tyrosyl-trna synthetase (cys-35, his-48, and lys-233) are not present in the human enzyme. this raises the question of whether the activation energy for the tyrosine activation step is higher for t ...200211856731
novel zinc-binding center and a temperature switch in the bacillus stearothermophilus l1 lipase.the bacterial thermoalkalophilic lipases optimally hydrolyze saturated fatty acids at elevated temperatures. they also have significant sequence homology with staphylococcal lipases, and both the thermoalkalophilic and staphylococcal lipases are grouped as the lipase family i.5. we report here the first crystal structure of the lipase family i.5, the structure of a thermoalkalophilic lipase from bacillus stearothermophilus l1 (l1 lipase) determined at 2.0-a resolution. the structure is in a clos ...200211859083
the effects of modifying the surface charge on the catalytic activity of a thermolysin-like protease.the impact of long range electrostatic interactions on catalysis in the thermolysin-like protease from bacillus stearothermophilus was studied by analyzing the effects of inserting or removing charges on the protein surface. various mutations were introduced at six different positions, and double-mutant cycle analysis was used to study the extent to which mutational effects were interdependent. the effects of single point mutations on the k(cat)/k(m) were non-additive, even in cases where the po ...200211859085
[comparative study of the m.bstf5i-1 and m.bstf5i-3 dna methyltransferases from the bacillus stearothermophilus f5 restriction-modification system].the bstf5i restriction-modification system from bacillus stearothermophilus f5, unlike all known restriction-modification systems, contains three genes encoding dna methyltransferases. in addition to revealing two dna methylases responsible for modification of adenine in different dna strands, it has been first shown that one bacterial cell has two dna methylases, m.bstf5i-1 and m.bstf5i-3, with similar substrate specificity. the boundaries of the gene for dna methyltransferase m.bstf5i-1 have b ...200211862704
on the interaction of ribosomal protein l5 with 5s rrna.ribosomal protein l5, a 5s rrna binding protein in the large subunit, is composed of a five-stranded antiparallel beta-sheet and four alpha-helices, and folds in a way that is topologically similar to the ribonucleprotein (rnp) domain [nakashima et al., rna 7, 692-701, 20011. the crystal structure of ribosomal protein l5 (bstl5) from bacillus stearothermophilus suggests that a concave surface formed by an anti-parallel beta-sheet and long loop structures are strongly involved in 5s rrna binding. ...200211866091
[m.bstf5i-4, the forth dna methyltransferase of bstf5i restriction-modification system from bacillus stearothermophilus f5].the fourth dna-methyltransferase of the bstf5i restriction-modification (rm) system from bacillus stearothermophilus f5 (m.bstf5i-4) was discovered, which modifies the adenine residue within the upper strand of the recognition site 5'-ggatg-3'/5'-catcc-3'. thus, unlike other known rm systems, the bstf5i rm system comprises four genes encoding dna-methyltransferases, three of which possess the same substrate specificity and methylate adenine within the 5'-ggatg sequence. the english version of th ...200211875980
samy, a novel mammalian reporter gene derived from bacillus stearothermophilus alpha-amylase.the bacillus stearothermophilus alpha-amylase (amys) is a heat-stable monomeric exoenzyme which catalyses random hydrolysis of 1,4-alpha-glucosidic linkages in polyglucosans. the bacillus alpha-amylase was engineered for use as an intracellular (amys(delta s)) as well as a secreted reporter protein (samy; secreted alpha-amylase) in mammalian cells. the 5' end of amys containing the prokaryotic secretion signal was either deleted (amys(delta s)) or replaced by a murine immunoglobulin secretion si ...200211814674
structure and mechanism of action of a cofactor-dependent phosphoglycerate mutase homolog from bacillus stearothermophilus with broad specificity phosphatase activity.the crystal structure of bacillus stearothermophilus phoe (originally termed yhfr), a broad specificity monomeric phosphatase with a molecular mass of approximately 24 kda, has been solved at 2.3 a resolution in order to investigate its structure and function. phoe, already identified as a homolog of a cofactor-dependent phosphoglycerate mutase, shares with the latter an alpha/beta/alpha sandwich structure spanning, as a structural excursion, a smaller subdomain composed of two alpha-helices and ...200211827481
catalytic activities of intracellular dimeric neopullulanase on cyclodextrin, acarbose and maltose.multi-substrate specificity of neopullulanase towards cyclodextrin, acarbose and maltose was investigated using a clone originating from bacillus stearothermophilus ima6503. the enzyme purified from escherichia coli harbouring the corresponding npla gene hydrolysed beta-cyclodextrin (beta-cd) to maltose and glucose. it exhibited substrate preference for beta-cd, starch and pullulan in the proportions of 10.4:1.2:1. the enzyme not only hydrolysed acarbose, an alpha-amylase inhibitor, to a pseudot ...200211834127
crystal structure of the bse634i restriction endonuclease: comparison of two enzymes recognizing the same dna sequence.crystal structures of type ii restriction endonucleases demonstrate a conserved common core and active site residues but diverse structural elements involved in dna sequence discrimination. comparative structural analysis of restriction enzymes recognizing the same nucleotide sequence might therefore contribute to our understanding of the structural diversity of specificity determinants within restriction enzymes. we have solved the crystal structure of the bacillus stearothermophilus restrictio ...200211842098
a functional interaction between the putative primosomal protein dnai and the main replicative dna helicase dnab in bacillus.in gram negative escherichia coli there are two well-characterised primosomal assembly processes, the pria- and dnaa-mediated cascades. the presence of pria and dnaa proteins in gram positive bacillus spp. supports the assumption that both the pria- and dnaa-mediated primosomal assembly cascades also operate in these organisms. however, the lack of sequence homology between the rest of the primosomal proteins indicates significant differences between these two bacterial species. central to the p ...200211842108
[isolation of endophytic bacteria in potato and test of antagonistic action to bacterial ring rot of potato].in this study, two hundred and forty bacterial strains were isolated from inner tissue of potato tubers collected from datong, taiyuan and inner mongolia autonomous regions. on the basis of antagonistic examination in vitro, fifty and five bacteria strains were characterized for antagonistic bacteria to ring rot of potato. it was 22.9 percentage of all bacteria strains. the biggest radius of suppression circle was 13 mm. nine strains were chosen for their suppression of bacterial ring rot, black ...200215346992
importance of an n-terminal extension in ribonuclease hii from bacillus stearothermophilus for substrate binding.the gene encoding ribonuclease hii from bacillus stearothermophilus was cloned and expressed in escherichia coli. the overproduced protein, bst-rnase hii, was purified and biochemically characterized. bst-rnase hii, which consists of 259 amino acid residues, showed the highest amino acid sequence identity (50.2%) to bacillus subtilis rnase hii. like b. subtilis rnase hii, it exhibited mn2+-dependent rnase h activity. it was, however, more thermostable than b. subtilis rnase hii. when the bst-rna ...200216233183
purification, characterization and gene cloning of thermostable o-acetyl-l-serine sulfhydrylase forming beta-cyano-l-alanine.a thermophilic and cyanide ion-tolerant bacterium, bacillus stearothermophilus cn3 isolated from a hot spring in japan, was found to produce thermostable beta-cyano-l-alanine synthase. the enzyme catalyzes the synthesis of beta-cyano-l-alanine from o-acetyl-l-serine and cyanide ions. the purified enzyme has a molecular mass of approximately 70 kda and consists of two identical subunits. it was stable in the ph range of 6.0 to 10.0 and up to 70 degrees c. the enzyme also catalyzes the synthesis o ...200316233442
synthesis of glycosyl glycerol by cyclodextrin glucanotransferases.glycerol was transglycosylated by cyclodextrin glucanotransferases using starch as a donor substrate. among the enzymes tested, those from geobacillus stearothermophilus and thermoanaerobacter sp. were suitable for the transglycosylation. several products were isolated and their structures were elucidated. they were composed of glucose and a series of a-1,4-linked maltooligosyl residues bound with glycerol. o-alpha-d-glucosyl-(1-->1)-glycerol and o-alpha-d-glucosyl-(1-->2)-glycerol were identifi ...200316233461
purification, characterization and gene cloning of thermostable o-acetyl-l-homoserine sulfhydrylase forming gamma-cyano-alpha-aminobutyric acid.a thermophilic and cyanide ion-tolerant bacterium, bacillus stearothermophilus cn3 isolated from a hot spring in japan, was found to produce thermostable gamma-cyano-alpha-aminobutyric acid synthase. the enzyme was purified and characterized. the purified enzyme has a molecular mass of approximately 180 kda and consists of four identical subunits. it was stable in the ph range of 6.0 to 10.5 and up to 60 degrees c. the enzyme catalyzed the gamma-replacement reaction of o-acetyl-l-homo-serine wit ...200316233482
prediction of the binding site on e1 in the assembly of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.the beta-subunit (e1beta) of the pyruvate decarboxylase (e1, alpha(2)beta(2)) component of the bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of pseudomonas putida and homo sapiens. based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of glu285 and the hydrophobic side chain of phe324 are of particular importance ...200314644451
near-infrared surface-enhanced-raman-scattering-mediated detection of single optically trapped bacterial spores.a novel methodology has been developed for the investigation of bacterial spores. specifically, this method has been used to probe the spore coat composition of two different bacillus stearothermophilus variants. this technique may be useful in many applications; most notably, development of novel detection schemes toward potentially harmful bacteria. this method would also be useful as an ancillary environmental monitoring system where sterility is of importance (i.e., food preparation areas as ...200314658146
inactivation of geobacillus stearothermophilus spores by high-pressure carbon dioxide treatment.high-pressure co2 treatment has been studied as a promising method for inactivating bacterial spores. in the present study, we compared this method with other sterilization techniques, including heat and pressure treatment. spores of bacillus coagulans, bacillus subtilis, bacillus cereus, bacillus licheniformis, and geobacillus stearothermophilus were subjected to co2 treatment at 30 mpa and 35 degrees c, to high-hydrostatic-pressure treatment at 200 mpa and 65 degrees c, or to heat treatment at ...200314660357
[a simple biological method for the fast identification of antibiotics].a method used to determine the quantitative and qualitative determination of antibiotics in blood, injury discharge, in human and animal urine as well as in foodstuffs (meat, milk and products made of them) is described. it is based on using obligate, thermophilic bacteria bacillus stearothermophilus, strains kk bkm b-213od and bkm b-718 with an optimum growth at 55-60 degrees c. meso- and psychrophilic bacteria cannot grow at the above temperature, therefore, the studied non-sterile material do ...200314971328
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