Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| use of fusion proteins and procaryotic display systems for delivery of hiv-1 antigens: development of novel vaccines for hiv-1 infection. | two non-pathogenic scaffolds (represented by the filamentous bacteriophage fd and the dihydrolipoyl acetyltransferase e2 protein of the bacillus stearothermophilus pyruvate dehydrogenase (pdh) complex) able to deliver human immunodeficiency virus (hiv)-1 antigenic determinants, were designed in our laboratories and investigated in controlled assay conditions. based on a modification of the phage display technology, we developed an innovative concept for a safe and inexpensive vaccine in which co ... | 2003 | 15049429 |
| a method of increasing test range and accuracy of bioindicators: geobacillus stearothermophilus spores. | spores of geobacillus stearothermophilus are very sensitive to changes in temperature. when validating sterilizing processes, the most common bioindicator (bi) is spores of geobacillus stearothermophilus atcc12980 and atcc7953 with about 10(6) spores /bi and a d121-value of about 2 minutes in water. because these spores of geobacillus stearothermophilus do not survive at a f0-value above 12 minutes, it has not been possible to evaluate the agreement between the biological f-value (f(bio)) and ph ... | 2003 | 14558699 |
| purification, crystallization and preliminary x-ray studies of a p-nitrophenylphosphatase from bacillus stearothermophilus. | thermostable p-nitrophenylphosphatase from bacillus stearothermophilus has been expressed in escherichia coli, purified and crystallized. the crystals belong to space group c(2), with unit-cell parameters a = 67.17 a, b = 57.84 a, c = 62.49 a and alpha = 90.0 degrees, beta = 95.4 degrees, gamma = 90.0 degrees. diffraction data were collected to 1.40 a resolution with a completeness of 94.7% (96.6% for the last shell), an r(fac) value of 0.074 (0.341) and an i/sigma (i) value of 30.1 (2.67). | 2003 | 14561143 |
| optimal determination of particle orientation, absolute hand, and contrast loss in single-particle electron cryomicroscopy. | a computational procedure is described for assigning the absolute hand of the structure of a protein or assembly determined by single-particle electron microscopy. the procedure requires a pair of micrographs of the same particle field recorded at two tilt angles of a single tilt-axis specimen holder together with the three-dimensional map whose hand is being determined. for orientations determined from particles on one micrograph using the map, the agreement (average phase residual) between par ... | 2003 | 14568533 |
| the geobacillus stearothermophilus v iscs gene, encoding cysteine desulfurase, confers resistance to potassium tellurite in escherichia coli k-12. | many eubacteria are resistant to the toxic oxidizing agent potassium tellurite, and tellurite resistance involves diverse biochemical mechanisms. expression of the iscs gene from geobacillus stearothermophilus v, which is naturally resistant to tellurite, confers tellurite resistance in escherichia coli k-12, which is naturally sensitive to tellurite. the g. stearothermophilus iscs gene encodes a cysteine desulfurase. a site-directed mutation in iscs that prevents binding of its pyridoxal phosph ... | 2003 | 13129955 |
| crystal structures of an archaeal class i cca-adding enzyme and its nucleotide complexes. | cca-adding enzymes catalyze the addition of cca onto the 3' terminus of immature trnas without using a nucleic acid template and have been divided into two classes based on their amino acid sequences. we have determined the crystal structures of a class i cca-adding enzyme from archeoglobus fulgidus (afcca) and its complexes with atp, ctp, or utp. although it and the class ii bacterial bacillus stearothermophilus cca enzyme (bstcca) have similar dimensions and domain architectures (head, neck, b ... | 2003 | 14636575 |
| crystal structure at 1.45 a resolution of alanine racemase from a pathogenic bacterium, pseudomonas aeruginosa, contains both internal and external aldimine forms. | the structure of the catabolic alanine racemase, dadx, from the pathogenic bacterium pseudomonas aeruginosa, reported here at 1.45 a resolution, is a dimer in which each monomer is comprised of two domains, an eight-stranded alpha/beta barrel containing the plp cofactor and a second domain primarily composed of beta-strands. the geometry of each domain is very similar to that of bacillus stearothermophilus alanine racemase, but the rotation between domains differs by about 15 degrees. this chang ... | 2003 | 14674749 |
| fluorescence based structural analysis of tryptophan analogue-amp formation in single tryptophan mutants of bacillus stearothermophilus tryptophanyl-trna synthetase. | the symmetrical dimer structure of tryptophanyl-trna synthetase is similar to that of tyrosyl-trna synthetase whose binding behavior and structural details have been elucidated in detail. the structure of both subunits after forming the intermediate tryptophanyl-amp has important implications for the binding of the cognate trna(trp). single tryptophan mutants of bacillus stearothermophilus tryptophanyl-trna synthetase have been constructed and expressed and used to probe structural changes in di ... | 2003 | 14674776 |
| purification, crystallization and preliminary x-ray analysis of the novel dead protein bstdead from bacillus stearothermophilus. | dead proteins are members of a large and diverse family of rna helicases that use energy from atp hydrolysis to unwind short regions of duplex rna. bstdead from bacillus stearothermophilus is a 436-amino-acid protein and a representative member of the dead protein family. in addition to the mechanistic core common to dead proteins, bstdead has a unique approximately 60-amino-acid c-terminal extension that may denote a specific biological role. bstdead has been crystallized in space group p4(1/3) ... | 2003 | 14501141 |
| evaluation of a low-temperature steam and formaldehyde sterilizer. | we evaluated a low-temperature steam and formaldehyde (ltsf) sterilizer based on the draft european standard pren 14180. microbiological tests were conducted on small and full loads using process challenge devices in five programs (p1-p5). with small loads all tests showed no growth of bacillus stearothermophilus (atcc7953) spores. however, positive cultures were observed with full-load tests using p5 (sterilization temperature, 50 degrees c). our data indicated that the load influenced the effi ... | 2003 | 14505609 |
| crystal structure and snapshots along the reaction pathway of a family 51 alpha-l-arabinofuranosidase. | high-resolution crystal structures of alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6, a family 51 glycosidase, are described. the enzyme is a hexamer, and each monomer is organized into two domains: a (beta/alpha)8-barrel and a 12-stranded beta sandwich with jelly-roll topology. the structures of the michaelis complexes with natural and synthetic substrates, and of the transient covalent arabinofuranosyl-enzyme intermediate represent two stable states in the double displacem ... | 2003 | 14517232 |
| molecular dynamics studies of alanine racemase: a structural model for drug design. | alanine racemase (alar) is a bacterial enzyme that catalyzes the interconversion of l- and d-alanine, which is an essential constituent of the peptidoglycan layer of the bacterial cell wall and requires pyridoxal 5'-phosphate (plp) as a cofactor. the enzyme is universal to bacteria, including mycobacteria, making it an attractive target for drug design. to investigate the effects of flexibility on the binding modes of the substrate and an inhibitor and to analyze how the active site is affected ... | 2003 | 14517907 |
| cloning, sequencing, and expression of the gene encoding the clostridium stercorarium alpha-galactosidase aga36a in escherichia coli. | the alpha-galactosidase gene aga36a of clostridium stercorarium f-9 was cloned, sequenced, and expressed in escherichia coli. the aga36a gene consists of 2,208 nucleotides encoding a protein of 736 amino acids with a predicted molecular weight of 84,786. aga36a is an enzyme classified in family 36 of the glycoside hydrolases and showed sequence similarity with some enzymes of family 36 such as geobacillus (formerly bacillus) stearothermophilus gala (57%) and agan (52%). the enzyme purified from ... | 2003 | 14586104 |
| trimeric subunit stoichiometry of the glutamate transporters from bacillus caldotenax and bacillus stearothermophilus. | catalysis of glutamate transport across cell membranes and coupling of the concentrative transport to sodium, proton, and potassium gradients are processes fundamental to organisms in all kingdoms of life. in bacteria, glutamate transporters participate in nutrient uptake, while in eukaryotic organisms, the transporters clear glutamate from the synaptic cleft. even though glutamate transporters are crucial to the viability of many life forms, little is known about their structure and quaternary ... | 2003 | 14596613 |
| use of the microorganism bacillus stearothermophilus as a model to evaluate toxicity of the lipophilic environmental pollutant endosulfan. | microorganisms are very powerful tools for the supply of information about the toxic effects of lipophilic compounds, since an impairment of cell growth usually occurs as a result of perturbations related, in most cases, with the partition of toxicants in membranes. the thermophilic eubacterium bacillus stearothermophilus has been used as a model system to identify alpha- and beta-endosulfan interactions with the membrane possibly related with the insecticide toxicity. two approaches have been p ... | 2003 | 14599450 |
| tamoxifen induces ultrastructural alterations in membranes of bacillus stearothermophilus. | tamoxifen (tam), a non-steroid antiestrogen, is the mostly used drug for chemotherapy and chemoprevention of breast cancer. however, the mechanisms by which tam inhibits cell proliferation in breast cancer are not fully understood. tam strongly incorporates in biomembranes and a variety of effects have been assigned to biophysical and biochemical interactions with membranes. therefore, a better understanding of the physicochemical basis of interaction of tam with biomembranes is essential to elu ... | 2003 | 14599454 |
| molecular mechanisms of the metabolite 4-hydroxytamoxifen of the anticancer drug tamoxifen: use of a model microorganism. | a strain of the thermophilic eubacterium bacillus stearothermophilus was used as a model system to identify membrane mediated cytotoxic effects of 4-hydroxytamoxifen, following previous studies with tamoxifen. with this experimental approach we attempted to further clarify tamoxifen and 4-hydroxytamoxifen membrane interactions often evoked as responsible for their multiple cellular effects. bacterial growth and the oxygen consumption rate provided quantitative data of the cytotoxic action of hyd ... | 2003 | 14599455 |
| m.bstf5i-2 and m.bstf5i-4 dna methyltransferases from bstf5i restriction-modification system of bacillus stearothermophilus f5. | the bstf5i restriction-modification system from bacillus stearothermophilus f5 includes four site-specific dna methyltransferases, thus differing from all known restriction-modification systems. here we demonstrated for the first time that one bacterial cell can possess two pairs of methylases with identical substrate specificities (methylases bstf5i-1 and bstf5i-3 recognize ggatg, whereas methylases bstf5i-2 and bstf5i-4 recognize catcc) that modify adenine residues on both dna strands. differe ... | 2003 | 14606938 |
| interactions of the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component in the assembly of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus. | the enzymes pyruvate decarboxylase (e1) and dihydrolipoyl dehydrogenase (e3) bind tightly but in a mutually exclusive manner to the peripheral subunit-binding domain (psbd) of dihydrolipoyl acetyltransferase in the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus. the use of directed mutagenesis, surface plasmon resonance detection and isothermal titration microcalorimetry revealed that several positively charged residues of the psbd, most notably arg135, play an importa ... | 2003 | 14622277 |
| phylogenetic conservation of rna secondary and tertiary structure in the trpedcfba operon leader transcript in bacillus. | expression of the trpedcfba operon of bacillus subtilis is regulated by transcription attenuation and translation control mechanisms. we recently determined that the b. subtilis trp leader readthrough transcript can adopt a mg(2+)-dependent tertiary structure that appears to interfere with trap-mediated translation control of trpe. in the present study, sequence comparisons to trp leaders from three other bacillus sp. were made, suggesting that rna secondary and tertiary structures are phylogene ... | 2003 | 14624006 |
| construction of recombinant s-layer proteins (rsbsa) and their expression in bacterial ghosts--a delivery system for the nontypeable haemophilus influenzae antigen omp26. | this study has investigated the feasibility of a combination of recombinant surface layer (s-layer) proteins and empty bacterial cell envelopes (ghosts) to deliver candidate antigens for a vaccine against nontypeable haemophilus influenzae (nthi) infections. the s-layer gene sbsa from bacillus stearothermophilus pv72 was used for the construction of fusion proteins. fusion of maltose binding protein (mbp) to the n-terminus of sbsa allowed expression of the s-layer in the periplasm of escherichia ... | 2003 | 12832124 |
| expression, purification, crystallization and preliminary crystallographic analysis of leishmania mexicana phosphoglycerate mutase. | bacterially expressed 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (ipgam) from leishmania mexicana with a six-his tag fused at its c-terminus was expressed from plasmid pet28a after iptg induction in escherichia coli cells and gave a yield of 20 mg of highly purified ipgam per litre of cell culture. crystals of the protein complexed with 3-phosphoglycerate were obtained by the hanging-drop method of vapour diffusion with peg 4000 as the precipitating agent in the presence of coba ... | 2003 | 12832797 |
| transglycosylation of glycosyl residues to cyclic tetrasaccharide by bacillus stearothermophilus cyclomaltodextrin glucanotransferase using cyclomaltodextrin as the glycosyl donor. | cyclomaltodextrin glucanotransferase (ec 2.4.1.19, abbreviated as cgtase) derived from bacillus stearothermophilus produced a series of transfer products from a mixture of cyclomaltohexaose and cyclic tetrasaccharide (cyclo[-->6)-alpha-d-glcp-(1-->3)-alpha-d-glcp-(1-->6)-alpha-d-glcp-(1-->3)-alpha-d-glcp-(1-->], cts). of the transfer products, only two components, saccharides a and d, remained and accumulated after digestion with glucoamylase. the total combined yield of the saccharides reached ... | 2003 | 12834287 |
| studies on bacillus stearothermophilus. part iii. transformation of testosterone. | bacillus stearothermophilus, a thermophilic bacterium isolated from the kuwaiti desert, produced a variety of monohydroxy androstene derivatives and an oxidized product when incubated with exogenous testosterone for 24 h at 65 degrees c. the major metabolite was identified as androst-4-en-3,17-dione while minor metabolites included 6 alpha-hydroxyandrost-4-en-3,17-dione, 6 beta-hydroxyandrost-4-en-3,17-dione, 6 alpha-hydroxytestosterone, and 6 beta-hydroxytestosterone. these metabolites were pur ... | 2003 | 12835920 |
| evaluation of screening test for detection of antimicrobial residues in ewe milk. | the effects of preservatives (potassium dichromate and sodium azide), heat treatment (untreated and 82 degrees c/10 min), and lactation stage upon the response of the microbial tests (brt aim and delvotest) utilized for the detection of residues of antimicrobial substances in ewe milk were examined. milk samples were collected from the morning milking of 50 manchega ewes every 2 wk, from 15 d postpartum until the end of lactation. a total of 2322 samples were analyzed by brt aim with prediffusio ... | 2003 | 12836929 |
| crystal structure of d-hydantoinase from burkholderia pickettii at a resolution of 2.7 angstroms: insights into the molecular basis of enzyme thermostability. | d-hydantoinase (d-hyd) is an industrial enzyme that is widely used in the production of d-amino acids which are precursors for semisynthesis of antibiotics, peptides, and pesticides. this report describes the crystal structure of d-hydantoinase from burkholderia pickettii (hyd(bp)) at a 2.7-a resolution. the structure of hyd(bp) consists of a core (alpha/beta)(8) triose phosphate isomerase barrel fold and a beta-sheet domain, and the catalytic active site consists of two metal ions and six highl ... | 2003 | 12837777 |
| interactions of antimicrobials in milk and their detection by the disk diffusion method and delvotest sp. | the combination of more than 2 different microbials might show interactions with various effects (synergistic, additive, antagonistic, or indifferent) on target microorgnisms. an objective of this paper was to evaluate the possible interactions of several antimicrobials--those used most frequently in the treatment of mastitis in clinical veterinary practice (beta-lactam antibiotics, aminoglycosides, peptides, other antibiotics, and sulfonamides)--and their consequences on detection limits. in th ... | 2003 | 12852571 |
| effects of domain dissection on the folding and stability of the 43 kda protein pgk probed by nmr. | the characterization of early folding intermediates is key to understanding the protein folding process. previous studies of the n-domain of phosphoglycerate kinase (pgk) from bacillus stearothermophilus combined equilibrium amide exchange data with a kinetic model derived from stopped-flow kinetics. together, these implied the rapid formation of an intermediate with extensive native-like hydrogen bonding. however, there was an absence of protection in the region proximal to the c-domain in the ... | 2003 | 12860138 |
| evidence of a thermal unfolding dimeric intermediate for the escherichia coli histone-like hu proteins: thermodynamics and structure. | the escherichia coli histone-like hu protein pool is composed of three dimeric forms: two homodimers, echualpha(2) and echubeta(2), and a heterodimer, echualphabeta. the relative abundance of these dimeric forms varies during cell growth and in response to environmental changes, suggesting that each dimer plays different physiological roles. here, differential scanning calorimetry and circular dichroism (cd) were used to study the thermal stability of the three e.coli hu dimers and show that eac ... | 2003 | 12875839 |
| crystallization and preliminary structure determination of the c-terminal truncated domain of the s-layer protein sbsc. | the c-terminal truncated form of the s-layer protein sbsc from geobacillus stearothermophilus, rsbsc(31-844), has been crystallized by the vapour-diffusion method using polyethylene glycol 6000 as a precipitating agent. the crystals diffract to 3 a resolution using synchrotron radiation and belong to space group p2(1), with unit-cell parameters a = 57.24, b = 98.91, c = 108.62 a, beta = 94.34 degrees. one molecule is present in the asymmetric unit, which corresponds to a solvent content of 65%. ... | 2003 | 12876353 |
| crystallization and preliminary x-ray diffraction data for the carboxylesterase est30 from bacillus stearothermophilus. | crystals have been grown of the carboxylesterase est30 from bacillus stearothermophilus by hanging-drop vapor diffusion using ammonium sulfate as precipitant. the crystals diffracted to better than 2.0 a resolution. x-ray diffraction data were reduced in space group c222(1), with unit-cell parameters a = 55.83, b = 58.15, c = 179.65 a. r(merge) was 0.038 for 17 449 independent reflections with a completeness of 85.1%. v(m) was calculated to be 2.43 a(3) da(-1), which suggested that there was one ... | 2003 | 12876355 |
| gene cloning, expression, and crystallization of a thermostable exo-inulinase from geobacillus stearothermophilus kp1289. | the gene ( inua) encoding exo-inulinase (ec 3.2.1.80) was cloned from the thermophilic geobacillus stearothermophilus ( bacillus stearothermophilus) kp 1289 growing at between 41 degrees c and 69 degrees c. the inua gene consisted of 1,482 bp encoding a protein of 493 amino acids. the deduced polypeptide of molecular mass ( m) 56,744 da showed strong sequence similarity to pseudomonas mucidolens exo-inulinase, bacillus subtilis levanase, paenibacillus polymyxa ( bacillus polymyxa) fructosyltrans ... | 2003 | 12883863 |
| production of tagatose by a recombinant thermostable l-arabinose isomerase from thermus sp. im6501. | a gene (thai) corresponding to l-arabinose isomerase from thermus strain im6501 was cloned by pcr. it comprised 1488 nucleotides and encoded a polypeptide of 496 residues with a predicted molecular weight of 56019 da. the deduced amino acid sequence had 96.8% identity with the l-arabinose isomerase of geobacillus stearothermophilus. recombinant thai with n-terminal hexa-tistidine tags was over-expressed in escherichia coli and purified by affinity chromatography using ni-nta resin. the purified ... | 2003 | 12889832 |
| engineering cyclodextrin glycosyltransferase into a starch hydrolase with a high exo-specificity. | cyclodextrin glycosyltransferase (cgtase) enzymes from various bacteria catalyze the formation of cyclodextrins from starch. the bacillus stearothermophilus maltogenic alpha-amylase (g2-amylase is structurally very similar to cgtases, but converts starch into maltose. comparison of the three-dimensional structures revealed two large differences in the substrate binding clefts. (i) the loop forming acceptor subsite +3 had a different conformation, providing the g2-amylase with more space at accep ... | 2003 | 12890607 |
| genetic polymorphism by rapd-pcr and phenotypic characteristics of isolated thermotolerant bacillus strains from hot spring sources. | the polymerase chain reaction (pcr) based random amplified polymorphic dna (rapd) assay, morphological, physiological, biochemical and antimicrobial susceptibility test methods have been evaluated for use in the taxonomy of isolated thermotolerant bacillus from jordanian hot springs, with specific reference to strains geobacillus stearothermophilus (atcc 12980), bacillus circulans (atcc 4513) and bacillus sphaericus (atcc 14577). a rapd assay has been optimized and is able to discriminate betwee ... | 2003 | 12901420 |
| a model for the thermal inactivation of micro-organisms. | to mathematically model published thermal inactivation data sets using an empirical model based on a double arrhenius function. | 2003 | 12911698 |
| conversion of a plp-dependent racemase into an aldolase by a single active site mutation. | alanine racemase (alr) [ec 5.1.1.1] from geobacillus stearothermophilus is a pyridoxal 5'-phosphate-dependent enzyme that catalyzes the first committed step in bacterial cell wall biosynthesis. it is converted to an aldolase upon replacement of tyr265, which normally serves as a catalytic base in the racemase reaction, with alanine. the y265a mutation increases catalytic efficiency for cleavage of beta-phenylserine to benzaldehyde and glycine by 2.3 x 105 fold as compared to the wild-type racema ... | 2003 | 12926923 |
| detailed kinetic analysis of a family 52 glycoside hydrolase: a beta-xylosidase from geobacillus stearothermophilus. | geobacillus stearothermophilus t-6 encodes for a beta-xylosidase (xynb2) from family 52 of glycoside hydrolases that was previously shown to hydrolyze its substrate with net retention of the anomeric configuration. xynb2 significantly prefers substrates with xylose as the glycone moiety and exhibits a typical bell-shaped ph dependence curve. binding properties of xylobiose and xylotriose to the active site were measured using isothermal titration calorimetry (itc). binding reactions were enthalp ... | 2003 | 12950180 |
| inhibition of beta-fructofuranosidases and alpha-glucosidases by synthetic thio-fructofuranoside. | a synthetic beta-thio-fructofuranoside of mercaptoethanol inhibited not only beta-fructofuranosidases but also alpha-glucosidases. the compound was hardly hydrolyzed by the glycosidases. the thio-fructoside competitively inhibited beta-fructofuranosidases from aspergillus niger, candida sp., and saccharomyces cerevisiae, but not arthrobacter beta-fructofuranosidase at all. sucrase activity of rat intestinal sucrase/isomaltase complex was also suppressed in the presence of the thio-fructoside. th ... | 2003 | 12951505 |
| automated image acquisition and processing using a new generation of 4k x 4k ccd cameras for cryo electron microscopic studies of macromolecular assemblies. | we have previously reported the development of autoem, a software package for semi-automated acquisition of data from a transmission electron microscope. in continuing efforts to improve the speed of structure determination of macromolecular assemblies by electron microscopy, we report here on the performance of a new generation of 4 k ccd cameras for use in cryo electron microscopic applications. we demonstrate that at 120 kv, and at a nominal magnification of 67000 x, power spectra and signal- ... | 2003 | 12972350 |
| direct chromatographic capture of enzyme from crude homogenate using immobilized metal affinity chromatography on a continuous supermacroporous adsorbent. | a continuous supermacroporous matrix has been developed allowing direct capture of enzyme from non-clarified crude cell homogenate at high flow-rates. the continuous supermacroporous matrix has been produced by radical co-polymerization of acrylamide, allyl glycidyl ether and n,n'-methylene-bis(acrylamide) which proceeds in aqueous solution of monomers frozen inside a column (cryo-polymerization). after thawing, the column contains a continuous matrix having interconnected pores of 10-100 microm ... | 2003 | 12597634 |
| site-directed mutagenesis of a loop at the active site of e1 (alpha2beta2) of the pyruvate dehydrogenase complex. a possible common sequence motif. | limited proteolysis of the pyruvate decarboxylase (e1, alpha2beta2) component of the pyruvate dehydrogenase (pdh) multienzyme complex of bacillus stearothermophilus has indicated the importance for catalysis of a site (tyr281-arg282) in the e1alpha subunit (chauhan, h.j., domingo, g.j., jung, h.-i. & perham, r.n. (2000) eur. j. biochem. 267, 7158-7169). this site appears to be conserved in the alpha-subunit of heterotetrameric e1s and multiple sequence alignments suggest that there are additiona ... | 2003 | 12603319 |
| induction of specific t-helper and cytolytic responses to epitopes displayed on a virus-like protein scaffold derived from the pyruvate dehydrogenase multienzyme complex. | the icosahedral protein scaffold (1.5mda) generated by self-assembly of the catalytic domains of the dihydrolipoyl acetyltransferase core of the pyruvate dehydrogenase multienzyme complex from bacillus stearothermophilus has been engineered to display 60 copies of one or more peptide epitopes on a single molecule (e2disp). an e2disp scaffold displaying pep23, a 15-residue b- and t-helper epitope from the reverse transcriptase of hiv-1, was able to induce a pep23-specific t-helper response in cel ... | 2003 | 12615447 |
| effect of carrier materials on the resistance of spores of bacillus stearothermophilus to gaseous hydrogen peroxide. | the testing of the h2o2 decontamination process using spores of bacillus stearothermophilus has gained widespread acceptance. usually, commercially available biological indicators (bis) with a specified resistance to h2o2 are challenged to qualify the process. the question arises whether the resistance of test spores is dependant on the type of carrier material and whether the resistance is representative for the system under test. the objective of the study is to quantify the effect of differen ... | 2003 | 12643502 |
| structure and dynamics of the dna-binding protein hu of b. stearothermophilus investigated by raman and ultraviolet-resonance raman spectroscopy. | the histone-like protein hu of bacillus stearothermophilus (hubst) is a 90-residue homodimer that binds nonspecifically to b dna. although the structure of the hubst:dna complex is not known, the proposed dna-binding surface consists of extended arms that project from an alpha-helical platform. here, we report raman and ultraviolet-resonance raman (uvrr) spectra diagnostic of subunit secondary structures and indicative of key side-chains lining the proposed dna-binding surface. raman conformatio ... | 2003 | 12649443 |
| secretory expression in escherichia coli and single-step purification of a heat-stable alkaline protease. | bacteriocin release proteins (brps) can be used for the release of heterologous proteins from the escherichia coli cytoplasm into the culture medium. the gene for a highly thermostable alkaline protease was cloned from bacillus stearothermophilus f1 by the polymerase chain reaction. the recombinant f1 protease was efficiently excreted into the culture medium using e. coli xl1-blue harboring two vectors: ptrchis bearing the protease gene and pjl3 containing the brps. both vectors contain the e. c ... | 2003 | 12651108 |
| repair of hydrolytic dna deamination damage in thermophilic bacteria: cloning and characterization of a vsr endonuclease homolog from bacillus stearothermophilus. | hydrolytic deamination of 5-methyl cytosine in double stranded dna results in formation of a t/g mismatch that-if left unrepaired-leads to a c-->t transition mutation in half of the progeny. in addition to several mismatch-specific glycosylases that have been found in both pro- and eukaryotes to channel this lesion into base excision repair by removing the t from the mismatch, vsr endonuclease from escherichia coli has been described which initiates repair by an endonucleolytic strand incision 5 ... | 2003 | 12655008 |
| high-resolution x-ray structure of the dna-binding protein hu from the hyper-thermophilic thermotoga maritima and the determinants of its thermostability. | the histone-like dna-binding proteins (hu) are a convenient model for studying factors affecting thermostability because of their relatively simple, easily comparable structures, their common function, and their presence in organisms of widely differing thermostability. we report the determination of the high-resolution structure (1.53 a) at 273 k and 100 k of the hu protein from the hyper-thermophilic eubacterium thermotoga maritima(hu tmar, t(m)=80.5 degrees c). the structural data presented c ... | 2003 | 12664263 |
| structure-function relationship in serine hydroxymethyltransferase. | serine hydroxymethyltransferase (shmt), a pyridoxal-5'-phosphate (plp)-dependent enzyme catalyzes the tetrahydrofolate (h(4)-folate)-dependent retro-aldol cleavage of serine to form 5,10-methylene h(4)-folate and glycine. the structure-function relationship of shmt was studied in our laboratory initially by mutation of residues that are conserved in all shmts and later by structure-based mutagenesis of residues located in the active site. the analysis of mutants showed that k71, y72, r80, d89, w ... | 2003 | 12686103 |
| optimization of a rf-generated cf4/o2 gas plasma sterilization process. | a sterilization process with the use of rf-generated (13.56 mhz) cf(4)/o(2) gas plasma was optimized in regards to power, flow rate, exposure time, and rf-system type. the dependency of the sporicidal effect on the spore inoculum positioning in the chamber of the rf systems was also investigated. dried bacillus stearothermophilus atcc 7953 endospores were used as test organisms. the treatments were evaluated on the basis of survival curves and corresponding d values. the only parameter found to ... | 2003 | 12687716 |
| dual coenzyme specificity of photosynthetic glyceraldehyde-3-phosphate dehydrogenase interpreted by the crystal structure of a4 isoform complexed with nad. | photosynthetic glyceraldehyde-3-phosphate dehydrogenase (gapdh) of spinacia oleracea belongs to a wide group of gapdhs found in most organisms displaying oxygenic photosynthesis, including cyanobacteria, green and red algae, and higher plants. as a major catalytic difference with respect to glycolytic gapdh, photosynthetic gapdh exhibits dual cofactor specificity toward pyridine nucleotides with a preference for nadp(h). here we report the crystal structure of nad-complexed recombinant a(4)-gapd ... | 2003 | 12705826 |
| multiple hydrogen kinetic isotope effects for enzymes catalyzing exchange with solvent: application to alanine racemase. | alanine racemase catalyzes the pyridoxal phosphate-dependent interconversion of the d- and l-isomers of alanine. previous studies have shown that the enzyme employs a two-base mechanism in which lys39 and tyr265 are the acid/base catalysts. it is thus possible that stereoisomerization of the external aldimine intermediates occurs through a concerted double proton transfer without the existence of a distinct carbanionic intermediate. this possibility was tested by the application of multiple kine ... | 2003 | 12718553 |
| effect of media on spore yield and thermal resistance of bacillus stearothermophilus. | the interference of eight components in the yield of sporulation and thermal resistance to moist heat (121 degrees c) of bacillus stearothermophilus spores suspended in 0.02 m calcium acetate solution and inoculated on paper strips previously treated with calcium acetate/calcium hydroxide was studied. the spore yield of 1.0 x 10(8)ml was developed at 62 degrees c in 17 media containing different concentrations of d-glucose, sodium chloride, l-glutamic acid, yeast extract, peptone, manganese sulf ... | 2003 | 12721452 |
| maltosyl-erythritol, a major transglycosylation product of erythritol by bacillus stearothermophilus maltogenic amylase. | this study was done to modify erythritol to change its physicochemical and sensory properties. erythritol, a four-carbon sugar alcohol, was transglycosylated by bacillus stearothermophilus maltogenic amylase with maltotriose as a donor molecule. the presence of various transglycosylation products of erythritol was confirmed by tlc and high performance ion exchange chromatography (hpic). the major transfer product was purified by gel filtration chromatography on bio-gel p-2. examination by lc-ms, ... | 2003 | 12723599 |
| insights into the catalytic mechanism of cofactor-independent phosphoglycerate mutase from x-ray crystallography, simulated dynamics and molecular modeling. | phosphoglycerate mutases catalyze the isomerization of 2 and 3-phosphoglycerates, and are essential for glucose metabolism in most organisms. here, we further characterize the 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (ipgm) from bacillus stearothermophilus by determination of a high-resolution (1.4a) crystal structure of the wild-type enzyme and the crystal structure of its s62a mutant. the mutant structure surprisingly showed the replacement of one of the two catalytically es ... | 2003 | 12729763 |
| identification of the catalytic residues in family 52 glycoside hydrolase, a beta-xylosidase from geobacillus stearothermophilus t-6. | beta-d-xylosidases (ec 3.2.1.37) are exo-type glycoside hydrolases that hydrolyze short xylooligosaccharides to xylose units. the enzymatic hydrolysis of the glycosidic bond involves two carboxylic acid residues, and their identification, together with the stereochemistry of the reaction, provides crucial information on the catalytic mechanism. two catalytic mutants of a beta-xylosidase from geobacillus stearothermophilus t-6 were subjected to detailed kinetic analysis to verify their role in ca ... | 2003 | 12738774 |
| a side reaction of alanine racemase: transamination of cycloserine. | alanine racemase (ec 5.1.1.1) catalyzes the interconversion of alanine enantiomers, and thus represents the first committed step involved in bacterial cell wall biosynthesis. cycloserine acts as a suicide inhibitor of alanine racemase and as such, serves as an antimicrobial agent. the chemical means by which cycloserine inhibits alanine racemase is unknown. through spectroscopic assays, we show here evidence of a pyridoxal derivative (arising from either isomer of cycloserine) saturated at the c ... | 2003 | 12741835 |
| dissecting dna-protein and protein-protein interactions involved in bacterial transcriptional regulation by a sensitive protein array method combining a near-infrared fluorescence detection. | the protein array methodology is used to study dna-protein and protein-protein interactions governing gene expression from the bacillus stearothermophilus pargco promoter-operator region. using probes labelled with near-infrared fluorescence dyes with exitation characteristics close to 700 or 800 nm, it is possible to detect signals from proteins (purified or non-purified in escherichia coli cell extracts) immobilised on a nitrocellulose membrane with a high sensitivity (almost 12 amol of a spot ... | 2003 | 12748944 |
| the phosphotriesterase gene opda in agrobacterium radiobacter p230 is transposable. | we report a transposase gene (tnpa) upstream of the opda phosphotriesterase gene of agrobacterium radiobacter p230, as well as inverted repeats indicative of insertion sequences, flanking the two genes. both the tnpa gene and the inverted repeats resemble the tn610 transposon from mycobacterium fortuitum. two additional putative open reading frames separate opda and tnpa with inferred translation products with similarity to two proteins encoded on the geobacillus stearothermophilus is5376 transp ... | 2003 | 12757939 |
| ahld, an n-acylhomoserine lactonase in arthrobacter sp., and predicted homologues in other bacteria. | quorum sensing is a signalling mechanism that controls diverse biological functions, including virulence, via n-acylhomoserine lactone (ahl) signal molecules in gram-negative bacteria. with the aim of isolating strains or enzymes capable of blocking quorum sensing by inactivating ahl, bacteria were screened for ahl degradation by their ability to utilize n-3-oxohexanoyl-l-homoserine lactone (ohhl) as the sole carbon source. among four isolates, strain ibn110, identified as arthrobacter sp., was ... | 2003 | 12777494 |
| the structure of glyceraldehyde 3-phosphate dehydrogenase from alcaligenes xylosoxidans at 1.7 a resolution. | the enzyme glyceraldehyde-3-phosphate dehydrogenase (gapdh) from the gram-negative denitrifying bacterial species alcaligenes xylosoxidans was purified and crystallized as a contaminant protein during purification of nitrous oxide reductase. this is the first structure of a gapdh from a denitrifying species. the crystal structure was solved at 1.7 a resolution by molecular replacement using the structure of gapdh from bacillus stearothermophilus as a starting model. the quality of the structure ... | 2003 | 12777799 |
| crystallization and preliminary x-ray analysis of a family 51 glycoside hydrolase, the alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6. | alpha-l-arabinofuranosidases (ec 3.2.1.55) are hemicellulases that cleave the glycosidic bond between l-arabinofuranoside side chains and various oligosaccharides. in this study, the first crystallization and preliminary x-ray analysis of the alpha-l-arabinofuranosidase from geobacillus stearothermophilus t-6 (abfa t-6), a family 51 glycoside hydrolase, is described. abfa t-6 is a hexameric protein consisting of six identical subunits of 502 amino acids and with a calculated molecular mass of 57 ... | 2003 | 12777810 |
| overexpression, purification, and characterization of the recombinant leucine aminopeptidase ii of bacillus stearothermophilus. | for expression of bacillus stearothermophilus ncib 8924 leucine aminopeptidase ii (lap ii) in escherichia coli regulated by a t5 promoter, the gene was amplified by polymerase chain reaction and cloned into expression vector pqe-32 to generate pqe-lapii. the his(6)-tagged enzyme was overexpressed in iptg-induced e. coli m15 (pqe-lapii) as a soluble protein and was purified to homogeneity by nickel-chelate chromatography to a specific activity of 425 u/mg protein with a final yield of 76%. the su ... | 2003 | 12783191 |
| reaction mechanism of the heterotetrameric (alpha2beta2) e1 component of 2-oxo acid dehydrogenase multienzyme complexes. | pyruvate decarboxylase (e1) catalyzes the first two reactions of the four involved in oxidative decarboxylation of pyruvate by the pyruvate dehydrogenase (pdh) multienzyme complex. it requires thiamin diphosphate to bring about the decarboxylation of pyruvate, which is followed by the reductive acetylation of a lipoyl group covalently bound to the n(6) amino group of a lysine residue in the second catalytic component, a dihydrolipoyl acetyltransferase (e2). replacement of two histidine residues ... | 2003 | 12795594 |
| inadequacy of manual cleaning for reprocessing single-use, triple-lumen sphinctertomes: simulated-use testing comparing manual with automated cleaning methods. | despite widespread reuse of single-use sphinctertomes, publications regarding the adequacy of reprocessing are conflicting and the cautery wire channel is seldom evaluated. our objective was to use thickened artificial test soil containing microorganisms to perform simulated-use tests combined with in-situ and destructive testing to evaluate cleaning efficacy and ethylene oxide sterilization of single-use triple lumen sphinctertomes. | 2003 | 12806356 |
| hu protein employs similar mechanisms of minor-groove recognition in binding to different b-dna sites: demonstration by raman spectroscopy. | the sequence isomers d(cgcaaatttgcg) and d(tcaaggccttga) form self-complementary duplexes that present distinct targets for binding of the homodimeric architectural protein hu of bacillus stearothermophilus (hubst). raman spectroscopy shows that although each duplex structure is of the b-dna type, there are subtle conformational dissimilarities between them, involving torsion angles of the phosphodiester backbone and the arrangements of stacked bases. each dna duplex forms a stable stoichiometri ... | 2003 | 12809494 |
| a rapd-based comparison of thermophilic bacilli from milk powders. | the similarity of strains of thermophilic geobacillus stearothermophilus (formerly bacillus stearothermophilus), anoxybacillus flavithermus (formerly bacillus flavothermus), bacillus licheniformis and bacillus subtilis isolated from separate milk powder production runs from multiple factories was examined using a random amplified polymorphic dna (rapd) protocol. as a result of the analysis of the rapd fingerprints and data relating to general growth and biochemical tests, over 98% of the 1470 is ... | 2003 | 12810270 |
| three-dimensional structure and substrate binding of bacillus stearothermophilus neopullulanase. | crystal structures of bacillus stearothermophilus trs40 neopullulanase and its complexes with panose, maltotetraose and isopanose were determined at resolutions of 1.9, 2.4, 2.8 and 3.2a, respectively. since the latter two carbohydrates are substrates of this enzyme, a deactivated mutant at the catalytic residue glu357-->gln was used for complex crystallization. the structures were refined at accuracies with r.m.s. deviations of bond lengths and bond angles ranging from 0.005a to 0.008a and 1.3 ... | 2003 | 12547200 |
| regulation of arginine biosynthesis in the psychropiezophilic bacterium moritella profunda: in vivo repressibility and in vitro repressor-operator contact probing. | we report the cloning of the arginine repressor gene from the psychropiezophilic gram-negative bacterium moritella profunda, the purification of its product (argr(mp)), the identification of the operator in the bipolar argecbfgh(a) operon, in vivo repressibility studies, and an in vitro analysis of the repressor-operator interaction, including binding to mutant and heterologous arginine operators. the argr(mp) subunit shows about 70% amino acid sequence identity with escherichia coli argr (argr( ... | 2003 | 12559906 |
| crystal structure of two ternary complexes of phosphorylating glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus with nad and d-glyceraldehyde 3-phosphate. | the crystal structure of the phosphorylating glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus was solved in complex with its cofactor, nad, and its physiological substrate, d-glyceraldehyde 3-phosphate (d-g3p). to isolate a stable ternary complex, the nucleophilic residue of the active site, cys(149), was substituted with alanine or serine. the c149a and c149s gapdh ternary complexes were obtained by soaking the crystals of the corresponding binary complexes (enz ... | 2003 | 12569100 |
| development of a genetic system for hyperthermophilic archaea: expression of a moderate thermophilic bacterial alcohol dehydrogenase gene in sulfolobus solfataricus. | the escherichia coli/sulfolobus solfataricus shuttle vector pexss was used as a cloning vehicle for the gene transfer and expression of two bacterial genes in sulfolobus solfataricus. the alcohol dehydrogenase (adh) from the moderate thermophilic bacillus stearothermophilus (strain lldr) and a mutagenised version encoding a less thermostable adh enzyme were the selected genes. s. solfataricus adh promoter and aspartate aminotransferase terminator were used to drive the heterologous gene expressi ... | 2003 | 12583906 |
| p but not r-axis interface is involved in cooperative binding of nad on tetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus. | homotetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus can be described as a dimer of dimers with three non-equivalent p, r, and q interfaces. in our previous study, negative cooperativity in nad binding to wild-type gapdh was interpreted according to the induced-fit model in terms of two independent dimers with two interacting binding sites in each dimer. two dimeric mutant gapdhs, i.e. y46g/s48g and d186g/e276g, were shown to exhibit po ... | 2003 | 12595262 |
| arginyl-trna synthetase with signature sequence kmsk from bacillus stearothermophilus. | argrs (arginyl-trna synthetase) belongs to the class i aarss (aminoacyl-trna synthetases), though the majority of argrs species lack the canonical kmsk sequence characteristic of class i aarss. a dna fragment of the argrs gene from bacillus stearothermophilus was amplified using primers designed according to the conserved regions of known argrss. through analysis of the amplified dna sequence and known trna(arg)s with a published genomic sequence of b. stearothermophilus, the gene encoding argrs ... | 2003 | 13678419 |
| structures of phosphate and trivanadate complexes of bacillus stearothermophilus phosphatase phoe: structural and functional analysis in the cofactor-dependent phosphoglycerate mutase superfamily. | bacillus stearothermophilus phosphatase phoe is a member of the cofactor-dependent phosphoglycerate mutase superfamily possessing broad specificity phosphatase activity. its previous structural determination in complex with glycerol revealed probable bases for its efficient hydrolysis of both large, hydrophobic, and smaller, hydrophilic substrates. here we report two further structures of phoe complexes, to higher resolution of diffraction, which yield a better and thorough understanding of its ... | 2003 | 12498792 |
| lysyl-trna synthetase from bacillus stearothermophilus: the trp314 residue is shielded in a non-polar environment and is responsible for the fluorescence changes observed in the amino acid activation reaction. | three trp variants of lysyl-trna synthetase from bacillus stearothermophilus, in which either one or both of the two trp residues within the enzyme (trp314 and trp332) were substituted by a phe residue, were produced by site-directed mutagenesis without appreciable loss of catalytic activity. the following two phenomena were observed with w332f and with the wild-type enzyme, but not with w314f: (1) the addition of l-lysine alone decreased the protein fluorescence of the enzyme, but the addition ... | 2003 | 12507472 |
| biophysical characterization of the entire bacterial surface layer protein sbsb and its two distinct functional domains. | the crystalline bacterial cell surface layer (s-layer) protein sbsb of geobacillus stearothermophilus pv72/p2 was dissected into an n-terminal part defined by the three consecutive s-layer homologous motifs and the remaining large c-terminal part. both parts of the mature protein were produced as separate recombinant proteins (rsbsb(1-178) and rsbsb(177-889)) and compared with the full-length form rsbsb(1-889) (rsbsb). evidence for functional and structural integrity of the two truncated forms w ... | 2004 | 14625307 |
| crystal structure of the bstdead n-terminal domain: a novel dead protein from bacillus stearothermophilus. | most cellular processes requiring rna structure rearrangement necessitate the action of asp-glu-ala-asp (dead) proteins. members of the family, named originally for the conserved dead amino acid sequence, are thought to disrupt rna structure and facilitate its rearrangement by unwinding short stretches of duplex rna. bstdead is a novel 436 amino acid representative of the dead protein family from bacillus stearothermophilus that contains all eight conserved motifs found in dead proteins and is h ... | 2004 | 14681586 |
| the solution structure of ribosomal protein l18 from bacillus stearothermophilus. | a medium resolution solution structure has been obtained for l18 from bacillus stearothermophilus (bstl18), a ribosomal protein that stabilizes the tertiary structure of 5s rrna and mediates its interaction with the rest of the large subunit. the n-terminal 22 amino acid residues of bstl18 are unstructured in solution. its remaining 98 residues form a globular domain that has the same topology as the globular domains of other l18s, but the orientation of helices is different. this conformational ... | 2004 | 14687565 |
| thermostability of multidomain proteins: elongation factors ef-tu from escherichia coli and bacillus stearothermophilus and their chimeric forms. | recombinant mesophilic escherichia coli (ec) and thermophilic bacillus stearothermophilus (bst) elongation factors ef-tus, their isolated g-domains, and six chimeric ef-tus composed of domains of either ef-tu were prepared, and their gdp/gtp binding activities and thermostability were characterized. bstef-tu and bstg-domain bound gdp and gtp with affinities in nanomolar and submicromolar ranges, respectively, fully comparable with those of ecef-tu. in contrast, the ecg-domain bound the nucleotid ... | 2004 | 14691225 |
| cloning and characterization of acetohydroxyacid synthase from bacillus stearothermophilus. | five genes from the ilv-leu operon from bacillus stearothermophilus have been sequenced. acetohydroxyacid synthase (ahas) and its subunits were separately cloned, purified, and characterized. this thermophilic enzyme resembles ahas iii of escherichia coli, and regulatory subunits of ahas iii complement the catalytic subunit of the ahas of b. stearothermophilus, suggesting that ahas iii is functionally and evolutionally related to the single ahas of gram-positive bacteria. | 2004 | 14702326 |
| disentangling the web of allosteric communication in a homotetramer: heterotropic inhibition of phosphofructokinase from bacillus stearothermophilus. | a strategy for isolating each of the four potentially unique heterotropic pairwise allosteric interactions that exist in the homotetramer phosphofructokinase from bacillus stearothermophilus is described. the strategy involves the construction of hybrid tetramers containing one wild-type subunit and three mutant subunits that have been modified to block binding of both the substrate, fructose 6-phosphate (fru-6-p), and the allosteric inhibitor, phospho(enol)pyruvate (pep). each type of binding s ... | 2004 | 14717614 |
| enhanced secretion of bacillus stearothermophilus l1 lipase in saccharomyces cerevisiae by translational fusion to cellulose-binding domain. | the secretion of bacillus stearothermophilus l1 lipase in saccharomyces cerevisiae was investigated by employing a fusion partner, a cellulose-binding domain (cbd) from trichoderma harzianum endoglucanase ii (theg). the cbd was connected to the n-terminal of l1 lipase through an endogenous linker peptide from theg. the expression cassette for the fusion protein in s. cerevisiae was constructed using the alpha-amylase signal peptide and the galactose-inducible gal10 promoter. secretion of cbd-lin ... | 2004 | 14740195 |
| antioxidative effects of glycosyl-ascorbic acids synthesized by maltogenic amylase to reduce lipid oxidation and volatiles production in cooked chicken meat. | glycosylated ascorbic acids were synthesized by using the transglycosylation activity of bacillus stearothermophilus maltogenic amylase with maltotriose to show effective antioxidative activity with enhanced oxidative stability. the modified ascorbic acids comprised mono- and di-glycosyl transfer products with an alpha-(1,6)-glycosidic linkage. the antioxidative effects of the glycosyl derivatives of ascorbic acid on the lipid oxidation of cooked chicken breast meat patties were compared, and th ... | 2004 | 14745161 |
| evaluation and establishing the performance of different screening tests for tetracycline residues in animal tissues. | four methods intended for screening muscle tissue for residues belonging to the tetracycline group were compared using artificially contaminated as well as incurred samples. two agar diffusion methods were studied: one with bacillus subtilis as a test strain, the second with bacillus cereus. two variants of each method were compared: thin plates for analysis of intact or minced meat, and thick plates for analysis of meat fluid. the thin plate variants could not be evaluated with artificially con ... | 2004 | 14754636 |
| dna-binding orientation and domain conformation of the e. coli rep helicase monomer bound to a partial duplex junction: single-molecule studies of fluorescently labeled enzymes. | the sf1 dna helicases are multi-domain proteins that can unwind duplex dna in reactions that are coupled to atp binding and hydrolysis. crystal structures of two such helicases, escherichia coli rep and bacillus stearothermophilus pcra, show that the 2b sub-domain of these proteins can be found in dramatically different orientations (closed versus open) with respect to the remainder of the protein, suggesting that the 2b domain is highly flexible. by systematically using fluorescence resonance e ... | 2004 | 14757053 |
| highly stable l-lysine 6-dehydrogenase from the thermophile geobacillus stearothermophilus isolated from a japanese hot spring: characterization, gene cloning and sequencing, and expression. | l-lysine dehydrogenase, which catalyzes the oxidative deamination of l-lysine in the presence of nad, was found in the thermophilic bacterium geobacillus stearothermophilus utb 1103 and then purified about 3,040-fold from a crude extract of the organism by using four successive column chromatography steps. this is the first report showing the presence of a thermophilic nad-dependent lysine dehydrogenase. the product of the enzyme catalytic activity was determined to be delta1-piperideine-6-carbo ... | 2004 | 14766574 |
| factors contributing to the occurrence of antimicrobial drug residues in kenyan milk. | the study investigated factors contributing to the occurrence of antimicrobial drug residues in milk within four major milk production districts in kenya. the frequency of contamination was studied among small- and large-scale dairy producers to determine if there were differences between the two types of producers. field samples (n = 1,600) were analyzed with the improved dutch tube diffusion test, a microbial inhibitor test (bacillus stearothermophilus). in total, 144 and 64 samples from small ... | 2004 | 14968978 |
| natural transformation of the thermophilic cyanobacterium thermosynechococcus elongatus bp-1: a simple and efficient method for gene transfer. | proteins derived from the thermophilic cyanobacterium thermosynechococcus elongatus bp-1, which performs plant-type oxygenic photosynthesis, are suitable for biochemical, biophysical and x-ray crystallographic studies. we found that t. elongatus displays natural transformation, and we established a simple and efficient protocol for transferring exogenous dnas into the organism's genome. we obtained transformants directly on selective agar plates without having to amplify them prior to plating. w ... | 2004 | 14639476 |
| crystal structures of geobacillus stearothermophilus alpha-glucuronidase complexed with its substrate and products: mechanistic implications. | alpha-glucuronidases cleave the alpha-1,2-glycosidic bond between 4-o-methyl-d-glucuronic acid and short xylooligomers as part of the hemicellulose degradation system. to date, all of the alpha-glucuronidases are classified as family 67 glycosidases, which catalyze the hydrolysis via the investing mechanism. here we describe several high resolution crystal structures of the alpha-glucuronidase (agua) from geobacillus stearothermophilus, in complex with its substrate and products. in the complex ... | 2004 | 14573597 |
| new-generation multicistronic expression platform: ptrident vectors containing size-optimized ires elements enable homing endonuclease-based cistron swapping into lentiviral expression vectors. | capitalizing on a proven multicistronic expression vector platform we have designed novel ptrident vectors which (1). enable coordinated expression of three desired transgenes, (2). are size-optimized, (3). take advantage of small highly efficient internal ribosome entry sites of the gtx or rbm3 type, (4). harbor various sites specific for homing endonucleases facilitating promoter/multicistronic expression unit/polyadenylation site swapping as well as (5). straightforward integration into human ... | 2004 | 15052637 |
| cloning of the o-acetylhomoserine sulfhydrylase gene from the ruminal bacterium selenomonas ruminantium hd4. | the o-acetylhomoserine sulfhydrylase (oahs) gene was cloned from a selenomonas ruminantium hd4 lambda zap ii genomic library by degenerative probe hybridization and complementation. sequence analysis revealed an 869-bp orf with a g + c content of 53%. the orf had significant homology with enzymes involved in homocysteine biosynthesis. a curablastn homology search showed that the orf has 63% nucleotide identity with the oahs of bacillus stearothermophilus, corynebacterium glutamicum, and acremoni ... | 2004 | 15057458 |
| s-layer glycan-specific loci on the chromosome of geobacillus stearothermophilus nrs 2004/3a and dtdp-l-rhamnose biosynthesis potential of g. stearothermophilus strains. | the approximately 16.5 kb surface layer (s-layer) glycan biosynthesis (slg) gene cluster of the gram-positive thermophile geobacillus stearothermophilus nrs 2004/3a has been sequenced. the cluster is located immediately downstream of the s-layer structural gene sgse and consists of 13 orfs that have been identified by database sequence comparisons. the cluster encodes dtdp-l-rhamnose biosynthesis (rml operon), required for building up the polyrhamnan s-layer glycan, as well as for assembly and e ... | 2004 | 15073305 |
| structures and analysis of highly homologous psychrophilic, mesophilic, and thermophilic adenylate kinases. | the crystal structures of adenylate kinases from the psychrophile bacillus globisporus and the mesophile bacillus subtilis have been solved and compared with that from the thermophile bacillus stearothermophilus. this is the first example we know of where a trio of protein structures has been solved that have the same number of amino acids and a high level of identity (66-74%) and yet come from organisms with different operating temperatures. the enzymes were characterized for their own thermal ... | 2004 | 15100224 |
| structure determination of the extracellular xylanase from geobacillus stearothermophilus by selenomethionyl mad phasing. | xylanases are hemicellulases that hydrolyze the internal beta-1,4-glycoside bonds of xylan. the extracellular thermostable endo-1,4-beta-xylanase (ec 3.2.1.8; xt6) produced by the thermophilic bacterium geobacillus stearothermophilus t-6 was shown to bleach pulp optimally at ph 9 and 338 k and was successfully used in a large-scale biobleaching mill trial. the xylanase gene was cloned and sequenced. the mature enzyme consists of 379 amino acids, with a calculated molecular weight of 43 808 da an ... | 2004 | 15103129 |
| nucleotide sequence of the cryptic plasmid ptt8 from thermus thermophilus hb8 and isolation and characterization of its high-copy-number mutant. | the complete nucleotide sequence of ptt8, a cryptic plasmid from thermus thermophilus hb8, was determined. ptt8 was 9328bp long and its g+c content was 69%. ptt8 contained eight putative open reading frames, three of which showed extensive similarities to the plasmid addiction proteins pasa and pasb of ptc-f14 and pam10.6, and the repa protein of the cole2-related plasmids, respectively. during the analysis of ptt8-based plasmid ppp442, which had been obtained during a promoter-screening experim ... | 2004 | 15109829 |
| cluster analysis of water molecules in alanine racemase and their putative structural role. | conservation of water molecules was identified by a cluster analysis of seven crystal structures of alanine racemase from bacillus stearothermophilus. a total of 47 clusters of consensus water sites were determined and found to be highly localized, as indicated by their low mobilities. these clusters are located in the region of the active sites as well as at the interface between the n-terminal domain (the alpha/beta-barrel) of the first monomer and the c-terminal domain of the second monomer. ... | 2004 | 15115851 |
| crystal structure and amide h/d exchange of binary complexes of alcohol dehydrogenase from bacillus stearothermophilus: insight into thermostability and cofactor binding. | the crystal structure of nad(+)-dependent alcohol dehydrogenase from bacillus stearothermophilus strain lld-r (htadh) was determined using x-ray diffraction data at a resolution of 2.35 a. the structure of homotetrameric htadh is highly homologous to those of bacterial and archaeal homotetrameric alcohol dehydrogenases (adhs) and also to the mammalian dimeric adhs. there is one catalytic zinc atom and one structural zinc atom per enzyme subunit. the enzyme was crystallized as a binary complex la ... | 2004 | 15122892 |
| crystal structure of ksga, a universally conserved rrna adenine dimethyltransferase in escherichia coli. | the bacterial enzyme ksga catalyzes the transfer of a total of four methyl groups from s-adenosyl-l-methionine (s-adomet) to two adjacent adenosine bases in 16s rrna. this enzyme and the resulting modified adenosine bases appear to be conserved in all species of eubacteria, eukaryotes, and archaebacteria, and in eukaryotic organelles. bacterial resistance to the aminoglycoside antibiotic kasugamycin involves inactivation of ksga and resulting loss of the dimethylations, with modest consequences ... | 2004 | 15136037 |
| a novel approach to specific allergy treatment: the recombinant fusion protein of a bacterial cell surface (s-layer) protein and the major birch pollen allergen bet v 1 (rsbsc-bet v 1) combines reduced allergenicity with immunomodulating capacity. | counterregulating the disease-eliciting th2-like immune response of allergen-specific th lymphocytes by fostering an allergen-specific th1-like response is a promising concept for future immunotherapy of type i allergy. the use of recombinant allergens combined with more functional adjuvants has been proposed. in this respect, we present a novel approach. the gene sequence encoding the major birch pollen allergen, bet v 1, was fused with the gene encoding the bacterial cell surface (s-layer) pro ... | 2004 | 15153479 |
| geobacillus stearothermophilus v ubie gene product is involved in the evolution of dimethyl telluride in escherichia coli k-12 cultures amended with potassium tellurate but not with potassium tellurite. | a 3.8-kb fragment of chromosomal dna of geobacillus stearothermophilus v cloned in psp72 (p1vh) confers resistance to potassium tellurite (k(2)teo(3)) and to potassium tellurate (k(2)teo(4)) when the encoded genes are expressed in escherichia coli k-12. the nt sequence of the cloned fragment predicts three orfs of 780, 399, and 600 bp, whose encoded protein products exhibit about 80% similarity with the sumt methyltransferase and the btur protein of bacillus megaterium, and with the ubie methylt ... | 2004 | 15164269 |