Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
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| gene replacement of adenylate kinase in the gram-positive thermophile geobacillus stearothermophilus disrupts adenine nucleotide homeostasis and reduces cell viability. | thermophilic bacteria are of great value for industry and research communities. unfortunately, the cellular processes and mechanisms of these organisms remain largely understudied. in the present study, we investigate how the inactivation of adenylate kinase (ak) affects the adenine nucleotide homeostasis of a gram-positive moderate thermophile, geobacillus stearothermophilus strain nub3621-r. ak plays a major role in the adenine nucleotide homeostasis of living cells and has been shown to be es ... | 2005 | 15647886 |
| evaluation of hydrogen peroxide vapour as a method for the decontamination of surfaces contaminated with clostridium botulinum spores. | the aim of this study was to evaluate the efficacy of hydrogen peroxide vapour (hpv) against spores of clostridium botulinum, for use as a method for decontaminating environments where this pathogen has been handled. spores were dried onto stainless steel slides and exposed to hpv in a sealed glovebox enclosure, transferred to a quenching agent at timed intervals during the exposure period, before survivors were cultured and enumerated. d-values were calculated from graphs of log10 survivors plo ... | 2005 | 15649542 |
| gene cloning, functional expression and secretion of the s-layer protein sgse from geobacillus stearothermophilus nrs 2004/3a in lactococcus lactis. | the ~93-kda surface layer protein sgse of geobacillus stearothermophilus nrs 2004/3a forms a regular crystalline array providing a nanopatterned matrix for the future display of biologically relevant molecules. lactococcus lactis nz9000 was established as a safe expression host for the controlled targeted production of sgse based on the broad host-range plasmid pnz124sph, into which the nisa promoter was introduced. sgse devoid of its signal peptide-encoding sequence was cloned into the new vect ... | 2005 | 15675069 |
| a thermodynamic study of mesophilic, thermophilic, and hyperthermophilic l-arabinose isomerases: the effects of divalent metal ions on protein stability at elevated temperatures. | to gain insight into the structural stability of homologous homo-tetrameric l-arabinose isomerases (ai), we have examined the isothermal guanidine hydrochloride (gdnhcl)-induced unfolding of ais from mesophilic bacillus halodurans (bhai), thermophilic geobacillus stearothermophilus (gsai), and hyperthermophilic thermotoga maritima (tmai) using circular dichroism spectroscopy. the gdnhcl-induced unfolding of the ais can be well described by a two-state reaction between native tetramers and unfold ... | 2005 | 15710423 |
| secretion of heterologous proteins in bacillus subtilis can be improved by engineering cell components affecting post-translocational protein folding and degradation. | to explore the potential to enhance secretion of heterologous proteins in bacillus subtilis by engineering cell factors affecting extracytoplasmic protein folding and degradation. | 2005 | 16033468 |
| tagatose production by immobilized recombinant escherichia coli cells containing geobacillus stearothermophilus l-arabinose isomerase mutant in a packed-bed bioreactor. | using immobilized recombinant escherichia coli cells containing geobacillus stearothermophilus l-arabinose isomerase mutant (gali 152), we found that the galactose isomerization reaction was maximal at 70 degrees c and ph 7.0. manganese ion enhanced galactose isomerization to tagatose. the immobilized cells were most stable at 60 degrees c and ph 7.0. the cell and substrate concentrations and dilution rate were optimal at 34 g/l, 300 g/l, and 0.05 h(-1), respectively. under the optimum condition ... | 2005 | 16080720 |
| toxicity of methoprene as assessed by the use of a model microorganism. | methoprene is an insect juvenile growth hormone mimic, commonly used as a pesticide. although widely used for the control of several pests, toxic effects on organisms of different phyla have been reported. these events triggered studies to clarify the mechanisms of toxicity of this insecticide putatively involved in ecological issues. here we show the effect of methoprene on the normal cell growth and viability of a strain of the thermophilic eubacterium bacillus stearothermophilus, previously u ... | 2005 | 16081242 |
| opposite roles of domains 2+3 of escherichia coli ef-tu and bacillus stearothermophilus ef-tu in the regulation of ef-tu gtpase activity. | the effect of noncatalytic domains 2+3 on the intrinsic activity and thermostability of the ef-tu gtpase center was evaluated in experiments with isolated domains 1 and six chimeric variants of mesophilic escherichia coli (ec) and thermophilic bacillus stearothermophilus (bst) ef-tus. the isolated catalytic domains 1 of both ef-tus displayed similar gtpase activities at their optimal temperatures. however, noncatalytic domains 2+3 of the ef-tus influenced the gtpase activity of domains 1 differe ... | 2005 | 16081328 |
| solution structure of the c1-subdomain of bacillus stearothermophilus translation initiation factor if2. | if2 is one of three bacterial translation initiation factors that are conserved through all kingdoms of life. it binds the 30s and 50s ribosomal subunits, as well as fmet-trnaf(met). after these interactions, fmet-trnaf(met) is oriented to the ribosomal p-site where the first amino acid of the nascent polypeptide, formylmethionine, is presented. the c-terminal domain of bacillus stearothermophilus if2, which is responsible for recognition and binding of fmet-trnaf(met), contains two structured m ... | 2005 | 16081655 |
| screening of catalytically active microorganisms for the synthesis of 6-modified purine nucleosides. | modified nucleosides can be prepared by microbial transglycosylation from cheaper nucleoside precursors using free or immobilised whole cells. an efficient screening method to find transglycosylation activity in microorganisms was developed for the synthesis of 6-modified purine nucleosides, such as 6-chloro-, 6-methoxy-, 6-iodo- and 6-mercaptopurine ribonucleoside. out of 100 microorganisms screened, bacillus stearothermophilus atcc 12980 was the best for this purpose. | 2005 | 16086256 |
| structure and hydride transfer mechanism of a moderate thermophilic dihydrofolate reductase from bacillus stearothermophilus and comparison to its mesophilic and hyperthermophilic homologues. | dihydrofolate reductase (dhfr) from a moderate thermophilic organism, bacillus stearothermophilus, has been cloned and expressed. physical characterization of the protein (bsdhfr) indicates that it is a monomeric protein with a molecular mass of 18,694.6 da (0.8), coincident with the mass of 18 694.67 da calculated from the primary sequence. determination of the x-ray structure of bsdhfr provides the first structure for a monomeric dhfr from a thermophilic organism, indicating a high degree of c ... | 2005 | 16114879 |
| associative mechanism for phosphoryl transfer: a molecular dynamics simulation of escherichia coli adenylate kinase complexed with its substrates. | the ternary complex of escherichia coli adenylate kinase (ecak) with its substrates adenosine monophosphate (amp) and mg-atp, which catalyzes the reversible transfer of a phosphoryl group between adenosine triphosphate (atp) and amp, was studied using molecular dynamics. the starting structure for the simulation was assembled from the crystal structures of ecak complexed with the bisubstrate analog diadenosine pentaphosphate (ap(5)a) and of bacillus stearothermophilus adenylate kinase complexed ... | 2005 | 15521058 |
| heat capacity-independent determination of differential free energy of stability between structurally homologous proteins. | under the assumption of equivalent heat capacity values, the differential free energy of stability for a pair of proteins midway between their thermal unfolding transition temperatures is shown to be independent of deltac(p) up to its cubic term in deltat(m). for model calculations reflecting the nearly 30 degrees c difference in t(m) for the adenylate kinases from the arctic bacterium bacillus globisporus and the thermophilic bacterium geobacillus stearothermophilus, the resultant error in esti ... | 2006 | 16125837 |
| crystal structure and structure-based mutational analyses of rnase hiii from bacillus stearothermophilus: a new type 2 rnase h with tbp-like substrate-binding domain at the n terminus. | ribonuclease hiii (bst-rnase hiii) from the moderate thermophile bacillus stearothermophilus is a type 2 rnase h but shows poor amino acid sequence identity with another type 2 rnase h, rnase hii. it is composed of 310 amino acid residues and acts as a monomer. bst-rnase hiii has a large n-terminal extension with unknown function and a unique active-site motif (dede), both of which are characteristics common to rnases hiii. to understand the role of these n-terminal extension and active-site res ... | 2006 | 16343535 |
| functional flexibility of bacillus stearothermophilus formamidopyrimidine dna-glycosylase. | the formamidopyrimidine-dna glycosylase (fpg) recognizes and eliminates efficiently 8-oxoguanine, an abundant mutagenic dna lesion. the x-ray structure of the inactive e3q mutant of fpg from bacillus stearothermophilus, complexed to an 8-oxog-containing dna, revealed a small peptide (called the alphaf-beta10 loop) involved in the recognition of the lesion via an interaction with the protonated n(7) atom. this region, which is disordered in the x-ray models where an abasic site-containing dna is ... | 2006 | 16857432 |
| thermal unfolding process of dihydrolipoamide dehydrogenase studied by fluorescence spectroscopy. | the thermal unfolding pathway for dihydrolipoamide dehydrogenase (lipdh) isolated from bacillus stearothermophilus was investigated focusing on the transient intermediate state characterized through time-resolved fluorescence studies. the decrease in ellipticity in the far uv region in the cd spectrum, the fluorescence spectral change of trp-91 and fad, and the thermal enzymatic inactivation curve consistently demonstrated that lipdh unfolded irreversibly on heat treatment at higher than 65 degr ... | 2006 | 16861247 |
| expression and characterization of the genes encoding azoreductases from bacillus subtilis and geobacillus stearothermophilus. | azoreductases have been characterized as enzymes that can decolorize azo dyes by reducing azo groups. in this study, genes encoding proteins having homology with the azoreductase gene of bacillus sp. oy1-2 were obtained from bacillus subtilis atcc6633, b. subtilis isw1214, and geobacillus stearotherophilus ifo13737 by polymerase chain reaction. all three genes encoded proteins with 174 amino acids. the deduced amino acid sequences of azoreductase homologs from b. subtilis isw1214, b. subtilis at ... | 2006 | 16861800 |
| evaluation of magnetic resonance for detection of bacterial contamination in low-acid, shelf-stable packaged soymilk. | this study evaluated magnetic resonance (mr) as a nondestructive method for detection of bacterial contamination in shelf-stable soymilk and cheese sauce. to accomplish this, individual 355-ml polymeric trays filled with soymilk and inoculated with bacillus stearothermophilus and bacillus subtilis (10(3) cfu) were incubated for up to 28 h at 55 degrees c and 62 h at 37 degrees c, respectively. mr relaxation times (t2) of these samples were then correlated with the bacterial growth as well as vis ... | 2006 | 16865902 |
| manual methods are suboptimal compared with automated methods for cleaning of single-use biopsy forceps. | most reusable biopsy forceps and all of the currently available single-use biopsy forceps do not have a port that allows fluid flow down the inner tubular shaft of the device. reusable biopsy forceps are widely used and reprocessed in healthcare facilities, and single-use biopsy forceps are reprocessed either in-house (eg, in canada and japan) or by third-party reprocessors (eg, in the united states). the objective of this study was to determine the cleaning efficacy of automated narrow-lumen so ... | 2006 | 16874645 |
| in vivo and in vitro evaluation of the efficacy of a peracetic acid-based disinfectant for decontamination of acrylic resins. | the purpose of this study was to assess the antimicrobial efficacy of a peracetic acid-based disinfectant for decontamination of heat-polymerized, chemically activated and microwave-polymerized acrylic resins. resin plates were contaminated in vivo upon intraoral use by 10 volunteers for 7 nights and slabs were contaminated in vitro by contact with bacillus subtilis and bacillus stearothermophilus. the contaminated acrylic resin specimens were immersed in a 0.2% peracetic acid-based disinfectant ... | 2006 | 16924337 |
| a novel synthetic mammalian promoter derived from an internal ribosome entry site. | introduction of specific mutations into a synthetic internal ribosome entry site (ires(gtx)) derived from the gtx homeodomain protein revealed additional transcriptional activity. this novel synthetic p(gtx) promoter exhibited consensus core promoter modules such as the initiator (inr) and the partial downstream promoter elements (dpe) and mediated high-level expression of a variety of transgenes including the human vascular endothelial growth factor 121 (vegf(121)), the human placental secreted ... | 2006 | 16924671 |
| production and characterization of thermostable alpha-amylase by thermophilic geobacillus stearothermophilus. | studies on the alpha-amylase-producing thermophilic bacterium isolated and identified from a hot spring in jordan and designated as geobacillus stearothermophilus jt2 were carried out. the optimum conditions for growth and enzyme production were ph 7 and 55 degrees c. the study of the kinetics of cellular growth indicated a mu(max) of 0.22/h, a k(s) of 1.2 g/l, a tau(d) of 3.15 h and a y(x/s) of 0.43 g cell/g starch. in addition, the activation energy for growth and death were estimated and foun ... | 2006 | 16927263 |
| internal contamination of air-driven low-speed handpieces and attached prophy angles. | in an in vitro crossover study, the authors investigated whether the interior of low-speed handpiece/prophy-angle systems becomes contaminated during operation and submersion into geobacillus stearothermophilus. | 2006 | 16946433 |
| computational studies of tryptophanyl-trna synthetase: activation of atp by induced-fit. | catalysis of amino acid activation by bacillus stearothermophilus tryptophanyl-trna synthetase involves three allosteric states: (1) open; (2) closed pre-transition state (prets); and (3) closed products (product). the interconversions of these states entail significant domain motions driven by ligand binding. we explore the application of molecular dynamics simulations to investigate ligand-linked conformational stability changes associated with this catalytic cycle. multiple molecular dynamics ... | 2006 | 16949606 |
| computational analysis of the mode of binding of 8-oxoguanine to formamidopyrimidine-dna glycosylase. | 8-oxoguanine (8og) is the most prevalent form of oxidative dna damage. in bacteria, 8og is excised by formamidopyrimidine glycosylase (fpg) as the initial step in base excision repair. to efficiently excise this lesion, fpg must discriminate between 8og and an excess of guanine in duplex dna. in this study, we explore the structural basis underlying this high degree of selectivity. two structures have been reported in which fpg is bound to dna, differing with respect to the position of the lesio ... | 2006 | 16953574 |
| adenoviral vector platform for transduction of constitutive and regulated tricistronic or triple-transcript transgene expression in mammalian cells and microtissues. | adenoviral particles can efficiently transduce a broad spectrum of cell types, so they are widely used in basic research and clinical trials. | 2006 | 16960915 |
| new insights into the glycosylation of the surface layer protein sgse from geobacillus stearothermophilus nrs 2004/3a. | the surface of geobacillus stearothermophilus nrs 2004/3a cells is covered by an oblique surface layer (s-layer) composed of glycoprotein subunits. to this s-layer glycoprotein, elongated glycan chains are attached that are composed of [-->2)-alpha-l-rhap-(1-->3)-beta-l-rhap-(1-->2)-alpha-l-rhap-(1-->] repeating units, with a 2-o-methyl modification of the terminal trisaccharide at the nonreducing end of the glycan chain and a core saccharide as linker to the s-layer protein. on sodium dodecyl s ... | 2006 | 16963578 |
| characterization of a new thermophilic spore photoproduct lyase from geobacillus stearothermophilus (splg) with defined lesion containing dna substrates. | the geobacillus stearothermophilus splg gene encodes a thermophilic spore photoproduct lyase (splg) that belongs to the family of radical s-adenosylmethionine (adomet) enzymes. the aerobically purified apo-splg forms a homodimer, which contains one [4fe-4s] cluster per monomer unit after reconstitution to the holoform. formation of the [4fe-4s] cluster was proven by quantification of the amount of iron and sulfur per homodimer and by uv and epr spectroscopy. the uv spectrum features a characteri ... | 2006 | 16968710 |
| the structural basis for the mutagenicity of o(6)-methyl-guanine lesions. | methylating agents are widespread environmental carcinogens that generate a broad spectrum of dna damage. methylation at the guanine o(6) position confers the greatest mutagenic and carcinogenic potential. dna polymerases insert cytosine and thymine with similar efficiency opposite o(6)-methyl-guanine (o6meg). we combined pre-steady-state kinetic analysis and a series of nine x-ray crystal structures to contrast the reaction pathways of accurate and mutagenic replication of o6meg in a high-fidel ... | 2006 | 17179038 |
| monomeric solution structure of the helicase-binding domain of escherichia coli dnag primase. | dnag is the primase that lays down rna primers on single-stranded dna during bacterial dna replication. the solution structure of the dnab-helicase-binding c-terminal domain of escherichia coli dnag was determined by nmr spectroscopy at near-neutral ph. the structure is a rare fold that, besides occurring in dnag c-terminal domains, has been described only for the n-terminal domain of dnab. the c-terminal helix hairpin present in the dnag c-terminal domain, however, is either less stable or abse ... | 2006 | 17010164 |
| destruction of spores on building decontamination residue in a commercial autoclave. | the u.s. environmental protection agency conducted an experiment to evaluate the effectiveness of a commercial autoclave for treating simulated building decontamination residue (bdr). the bdr was intended to simulate porous materials removed from a building deliberately contaminated with biological agents such as bacillus anthracis (anthrax) in a terrorist attack. the purpose of the tests was to assess whether the standard operating procedure for a commercial autoclave provided sufficiently robu ... | 2006 | 17012597 |
| [bacterial dna alteration by plasma generated atomic nitrogen. real-time pcr detection contribution]. | bacterial sterilization by the technology of plasma in post-discharge shows a growing interest. the main appeal of this new process resides in its action at dry and low temperature (60 degrees c). this technology would be therefore useful for the complex medical equipment, sensitive to the oxidization, humidity and/or requiring a temperature lower than 60 degrees c. the objective of this survey is to demonstrate the activity of an atomic flux emanating a plasma of pure molecular nitrogen on the ... | 2006 | 17027193 |
| stability of the 'l12 stalk' in ribosomes from mesophilic and (hyper)thermophilic archaea and bacteria. | the ribosomal stalk complex, consisting of one molecule of l10 and four or six molecules of l12, is attached to 23s rrna via protein l10. this complex forms the so-called 'l12 stalk' on the 50s ribosomal subunit. ribosomal protein l11 binds to the same region of 23s rrna and is located at the base of the 'l12 stalk'. the 'l12 stalk' plays a key role in the interaction of the ribosome with translation factors. in this study stalk complexes from mesophilic and (hyper)thermophilic species of the ar ... | 2006 | 17053098 |
| post-transcriptional regulation of the xyna expression by a novel mrna binding protein, xaif. | xaif, a novel 32kda protein encoded by the orf located in the immediate downstream of the xyna gene of bacillus stearothermophilus no. 236, was identified to be the xylanase-specific trans-activator. in this study, the positive effect of xaif was confirmed to be xylanase-specific, and the results from northern blot and in vitro transcription assays showed that the xaif increased the xyna transcripts at post-transcriptional step. moreover, analysis of the mrna decay rate led to the assertion that ... | 2006 | 17055461 |
| [effects on performance of high-speed dental handpieces subjected to autoclaving]. | to investigate the effects on performances of high-speed dental handpieces subjected to autoclaving. | 2006 | 17097010 |
| structural insights into the mechanism of the plp synthase holoenzyme from thermotoga maritima. | pyridoxal 5'-phosphate (plp) is the biologically active form of vitamin b6 and is an important cofactor for several of the enzymes involved in the metabolism of amine-containing natural products such as amino acids and amino sugars. the plp synthase holoenzyme consists of two subunits: yaad catalyzes the condensation of ribulose 5-phosphate, glyceraldehyde-3-phosphate, and ammonia, and yaae catalyzes the production of ammonia from glutamine. here we describe the structure of the plp synthase com ... | 2006 | 17144654 |
| comparison of two radio-frequency plasma sterilization processes using microspot evaluation of microbial inactivation. | in this study, we evaluated gas plasma surface sterilization methods in a specific sterilizer. we have introduced a new monitoring method using 0.4 microm pore size membranes, which in this study gave the information corresponding to 3000 exposed biological indicators per treatment cycle. this enabled us to compare the fraction of inoculates that showed no growth after exposure for 30 different locations in the chamber, and hereby identify weak and strong spots in the chamber with regard to spor ... | 2006 | 16362959 |
| cloning, purification and biochemical characterization of metallic-ions independent and thermoactive l-arabinose isomerase from the bacillus stearothermophilus us100 strain. | the araa gene encoding l-arabinose isomerase from bacillus stearothermophilus us100 strain was cloned, sequenced and over-expressed in e. coli. this gene encodes a 496-amino acid protein with a calculated molecular weight of 56.161 kda. its amino acid sequence displays the highest identity with l-ai from thermus sp. im6501 (98%) and that of geobacillus stearothermophilus t6 (97%). according to sds-page analysis, under reducing and non-reducing conditions, the recombinant enzyme has an apparent m ... | 2006 | 16386851 |
| expression of the ubie gene of geobacillus stearothermophilus v in escherichia coli k-12 mediates the evolution of selenium compounds into the headspace of selenite- and selenate-amended cultures. | the ubie gene of geobacillus stearothermophilus v, with its own promoter, was cloned and introduced into escherichia coli. the cloned gene complemented the ubie gene deficiency of e. coli an70. in addition, the expression of this gene in e. coli jm109 resulted in the evolution of volatile selenium compounds when these cells were grown in selenite- or selenate-amended media. these compounds were dimethyl selenide and dimethyl diselenide. | 2006 | 16391146 |
| structure and reactivity of a thermostable prokaryotic nitric-oxide synthase that forms a long-lived oxy-heme complex. | in an effort to generate more stable reaction intermediates involved in substrate oxidation by nitric-oxide synthases (noss), we have cloned, expressed, and characterized a thermostable nos homolog from the thermophilic bacterium geobacillus stearothermophilus (gsnos). as expected, gsnos forms nitric oxide (no) from l-arginine via the stable intermediate n-hydroxy l-arginine (noha). the addition of oxygen to ferrous gsnos results in long-lived heme-oxy complexes in the presence (soret peak 427 n ... | 2006 | 16407211 |
| in the bacillus stearothermophilus dnab-dnag complex, the activities of the two proteins are modulated by distinct but overlapping networks of residues. | we demonstrate the primase activity of bacillus stearothermophilus dnag and show that it initiates at 3'-atc-5' and 3'-att-5' sites synthesizing primers that are 22 or 23 nucleotides long. in the presence of the helicase dnab the size distribution of primers is different, and a range of additional smaller primers are also synthesized. nine residues from the n- and c-terminal domains of dnab, as well as its linker region, have been reported previously to affect this interaction. in bacillus stear ... | 2006 | 16452437 |
| biosynthesis of a thermostable gellan lyase by newly isolated and characterized strain of geobacillus stearothermophilus 98. | the thermophilic strain able to degrade gellan was isolated from bulgarian hot spring. according to its morphological and biochemical properties and by partial sequencing of its 16s rdna, it was classified as geobacillus stearothermophilus. it grew in a synthetic medium with gellan as the only carbon source with a specific growth rate of 0.69 h(-1) and generation time of 60 min. the strain produced thermostable gellan lyase extracellularly during exponential phase. its synthesis was inducible; t ... | 2006 | 16482399 |
| comparison of hydrogen peroxide and peracetic acid as isolator sterilization agents in a hospital pharmacy. | the efficacy of hydrogen peroxide and peracetic acid as isolator sterilization agents was compared. | 2006 | 16484519 |
| molecular cloning and characterization of an enzyme hydrolyzing p-nitrophenyl alpha-d-glucoside from bacillus stearothermophilus sa0301. | bacillus stearothermophilus sa0301 produces an extracellular oligo-1,6-glucosidase (bso16g) that also hydrolyzes p-nitrophenyl alpha-d-glucoside (tonozuka et al., j. appl. glycosci., 45, 397-400 (1998)). we cloned a gene for an enzyme hydrolyzing p-nitrophenyl alpha-d-glucoside, which was different from the one mentioned above, from b. stearothermophilus sa0301. the k(0)/k(m) values of bso16g for isomaltotriose and isomaltose were 13.2 and 1.39 s(-1).mm(-1) respectively, while the newly cloned e ... | 2006 | 16495668 |
| crystallization and preliminary x-ray diffraction studies of two thermostable alpha-galactosidases from glycoside hydrolase family 36. | alpha-galactosidases from thermophilic organisms have gained interest owing to their applications in the sugar industry. the alpha-galactosidases agaa, agab and agaa a355e mutant from geobacillus stearothermophilus have been overexpressed in escherichia coli. crystals of agab and agaa a355e have been obtained by the vapour-diffusion method and synchrotron data have been collected to 2.0 and 2.8 a resolution, respectively. crystals of agab belong to space group i222 or i2(1)2(1)2(1), with unit-ce ... | 2006 | 16511274 |
| crystallization and preliminary crystallographic analysis of the catechol 2,3-dioxygenase pheb from bacillus stearothermophilus br219. | class ii extradiol-cleaving catecholic dioxygenase, a key enzyme of aromatic compound degradation in bacteria, cleaves the aromatic ring of catechol by adding two o atoms. pheb is one of the class ii extradiol-cleaving catecholic dioxygenases and shows a high substrate specificity for catechol derivatives, which have one aromatic ring. in order to reveal the mechanism of the substrate specificity of pheb, pheb has been crystallized by the hanging-drop vapour-diffusion method using peg 4000 as a ... | 2006 | 16511281 |
| an enzymatically produced novel cyclomaltopentaose cyclized from amylose by an alpha-(1-->6)-linkage, cyclo-{-->6)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->}. | a bacterial strain am7, isolated from soil and identified as bacillus circulans, produced two kinds of novel cyclic oligosaccharides. the cyclic oligosaccharides were produced from amylose using a culture supernatant of the strain as the enzyme preparation. the major product was a cyclomaltopentaose cyclized by an alpha-(1-->6)-linkage, cyclo-{-->6)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->}. the other minor product was cyclomaltohexao ... | 2006 | 16545346 |
| efficacy of supplementation of alpha-amylase-producing bacterial culture on the performance, nutrient use, and gut morphology of broiler chickens fed a corn-based diet. | a trial was conducted to evaluate the efficacy of an escherichia coli strain producing alpha-amylase of bacillus stearothermophilus on growth performance, nutrient use, and the morphology of the small intestine of broilers fed a corn-based diet. one hundred thirty-five 1-d-old chicks (cobb 500) were randomly divided into 3 groups and treated as follows: (i) basal diet (control); (ii) basal diet and water supplemented with an e. coli strain that produced amylase, and (iii) basal diet and water su ... | 2006 | 16553283 |
| nad(h) recycling activity of an engineered bifunctional enzyme galactose dehydrogenase/lactate dehydrogenase. | a chimeric bifunctional enzyme composing of galactose dehydrogenase (galdh; from pseudomonas fluorescens) and lactate dehydrogenase (ldh; from bacillus stearothermophilus) was successfully constructed. the chimeric galdh/ldh possessed dual characteristics of both galactose dehydrogenase and lactate dehydrogenase activities while exhibiting hexameric rearrangement with a molecular weight of approximately 400 kda. in vitro observations showed that the chimeric enzyme was able to recycle nad with a ... | 2006 | 16585948 |
| harnessing asymmetrical substrate recognition by thermostable endov to achieve balanced linear amplification in multiplexed snp typing. | multiplexed amplification of specific dna sequences, by pcr or by strand-displacement amplification, is an intrinsically biased process. the relative abundance of amplified dna can be altered significantly from the original representation and, in extreme cases, allele dropout can occur. in this paper, we present a method of linear amplification of dna that relies on the cooperative, sequence-dependent functioning of the dna mismatch-repair enzyme endonuclease v (endov) from thermotoga maritima ( ... | 2006 | 16609704 |
| structural perturbations induced in linear and circular dna by the architectural protein hu from bacillus stearothermophilus. | hu is a small dna-binding protein of eubacteria that is believed to induce or stabilize bending of the double helix and mediate nucleoid compaction in vivo. although hu does not bind preferentially to specific dna sequences, it is known to have high affinity for dna sites containing structural anomalies, such as unpaired or mismatched bases, nicks, and four-way junctions. we have employed raman spectroscopy to further investigate the structural basis of hu-dna recognition in solution. experiment ... | 2006 | 16618125 |
| analysis of reduction of geobacillus stearothermophilus spores treated with high hydrostatic pressure and mild heat in milk buffer. | our unpublished experimental results of fractional factorial experiments showed that the significant external factors affecting high pressure processing (hpp) inactivation were pressure, temperature and pressure holding time. based on these results, response surface methodology (rsm) was employed in the present work and a quadratic equation for hpp inactivation was built. by analyzing the response surface plots and their corresponding contour plots as well as solving the quadratic equation, the ... | 2006 | 16621090 |
| the structure of an inverting gh43 beta-xylosidase from geobacillus stearothermophilus with its substrate reveals the role of the three catalytic residues. | beta-d-xylosidases are glycoside hydrolases that catalyze the release of xylose units from short xylooligosaccharides and are engaged in the final breakdown of plant cell-wall hemicellulose. here we describe the enzyme-substrate crystal structure of an inverting family 43 beta-xylosidase, from geobacillus stearothermophilus t-6 (xynb3). each xynb3 monomeric subunit is organized in two domains: an n-terminal five-bladed beta-propeller catalytic domain, and a beta-sandwich domain. the active site ... | 2006 | 16631196 |
| evaluation of a bacillus stearothermophilus tube test as a screening tool for anticoccidial residues in poultry. | a bacillus stearothermophilus var. calidolactis c953 tube test was evaluated for its ability in detecting the residue of selected anticoccidial drugs in poultry, specially sulfamethazine, furazolidone, and amprolium. various concentrations of each drug were injected into chicken liver and kidney tissues and these tissues were tested to determine the drug detection limits for each drug. the detection limit was defined as the drug concentration at which 95 % of the test results were interpreted as ... | 2006 | 16645344 |
| cloning and characterization of the hpr kinase/phosphorylase gene from bacillus stearothermophilus no. 236. | the bacillus stearothermophilus no. 236 gene encoding the bifunctional enzyme hprk/p, the key regulator of carbon catabolite repression/activation (ccr/cca) in most gram-positive bacteria, was cloned and the (his)(6)-tagged gene product was characterized in detail. the nucleotide sequence of the hprk/p gene corresponded to an open reading frame of 951 bp that encoded a polypeptide of 316 amino acid residues with a calculated molecular mass of 35,458 da. the deduced amino acid sequence of the b. ... | 2006 | 16717408 |
| cadmium ion biosorption by the thermophilic bacteria geobacillus stearothermophilus and g. thermocatenulatus. | this study reports surface complexation models (scms) for quantifying metal ion adsorption by thermophilic microorganisms. in initial cadmium ion toxicity tests, members of the genus geobacillus displayed the highest tolerance to cdcl2 (as high as 400 to 3,200 microm). the thermophilic, gram-positive bacteria geobacillus stearothermophilus and g. thermocatenulatus were selected for further electrophoretic mobility, potentiometric titration, and cd2+ adsorption experiments to characterize cd2+ co ... | 2006 | 16751511 |
| requirements and infection prophylaxis for internally cooled implant drills. | implant site preparation is crucially important to long-term success. heat generation during drilling is unfavourable, since bone is relatively susceptible to heat, depending on its vascularisation and microstructure. numerous factors such as drilling pressure, number of revolutions, drill design, wear and material, drilling depth and cooling influence heat generation. internally cooled drills are, therefore, increasingly used, even though the improved cooling effect compared to conventional ext ... | 2006 | 16783733 |
| synthesis and stereochemical assignment of dna spore photoproduct analogues. | investigation of the dna repair process performed by the spore photoproduct (sp) lyase repair enzyme is strongly hampered by the lack of defined substrates needed for detailed enzymatic studies. the problem is particularly severe because the repair enzyme belongs to the class of strongly oxygen-sensitive radical (s)-adenosylmethionine (sam) enzymes, which are notoriously difficult to handle. we report the synthesis of the spore photoproduct analogues 1 a and 1 b, which have open backbones and ar ... | 2006 | 16789031 |
| structure-based model of the stepping motor of pcra helicase. | dna helicases are ubiquitous molecular motors involved in cellular dna metabolism. they move along single-stranded dna (ssdna) and separate duplex dna into its component strands, utilizing the free energy from atp hydrolysis. the pcra helicase from bacillus stearothermophilus translocates as a monomer progressively from the 3' end to the 5' end of ssdna and is one of the smallest motor proteins structurally known in full atomic detail. using high-resolution crystal structures of the pcra-dna com ... | 2006 | 16815905 |
| identification of glutamate residues important for catalytic activity of bacillus stearothermophilus leucine aminopeptidase ii. | each of four conserved glutamate residues of bacillus stearothermophilus leucine aminopeptidase ii (bslapii) was replaced with aspartate, lysine, and leucine respectively by site-directed mutagenesis. the over-expressed wild-type and mutant enzymes were purified to homogeneity by nickel-chelate chromatography and the molecular mass of the subunit was determined to be 44.5 kda by sds-page. the specific activity for the glu-316 and glu-340 mutants was completely abolished, while glu-249 mutants sh ... | 2006 | 16820970 |
| antibodies to inactive conformations of glyceraldehyde-3-phosphate dehydrogenase inactivate the apo- and holoforms of the enzyme. | polyclonal antibodies produced after the immunization of a rabbit with glyceraldehyde-3-phosphate dehydrogenase (gapdh) from bacillus stearothermophilus were used to isolate two types of antibodies interacting with different non-native forms of the antigen. type i antibodies were purified using sepharose-bound apo-gapdh that was treated with glutaraldehyde to stabilize the enzyme in the tetrameric form. type ii antibodies were isolated using immobilized denatured monomers of the enzyme. it was s ... | 2006 | 16827661 |
| characterization of a mutated geobacillus stearothermophilus l-arabinose isomerase that increases the production rate of d-tagatose. | characterization of a mutated geobacillus stearothermophilus l-arabinose isomerase used to increase the production rate of d-tagatose. | 2006 | 16834609 |
| heterologous protein production capacity of mammalian cells cultivated as monolayers and microtissues. | a precise understanding of processes managing heterologous protein production in vitro and in vivo is essential for the manufacture of sophisticated biopharmaceuticals as well as for future gene therapy and tissue engineering initiatives. capitalizing on the gravity-enforced self-assembly of monodispersed cells into coherent (multicellular) microtissues we studied heterologous protein production of microtissues and monolayers derived from cell lines and primary cells engineered/transduced for (i ... | 2006 | 16255048 |
| inactivation of bacillus spores by the combination of moderate heat and low hydrostatic pressure in ketchup and potage. | the combination effect of moderate heat and low hydrostatic pressure (mhp) on the reduction of bacillus subtilis, bacillus coagulans and geobacillus stearothermophilus spores in food materials (potage and ketchup) was investigated. these bacterial spores were suspended in potage (ph 7), acidified potage (ph 4), neutralized ketchup (ph 7) and ketchup (ph 4). the suspensions were treated with and without pressure (100 mpa) and temperatures of 65-85 degrees c for 3 to 12 h. the bacterial spores wer ... | 2006 | 16260058 |
| thermostability enhancement and change in starch hydrolysis profile of the maltohexaose-forming amylase of bacillus stearothermophilus us100 strain. | the implications of asn315 and val450 in the atypical starch hydrolysis profile of bacillus stearothermophilus amy (a-amylase) us100 have been suggested previously [ben ali, mhiri, mezghani and bejar (2001) enzyme microb. tech. 28, 537-542]. in order to confirm this hypothesis, three mutants were generated. of these two have a single mutation, n315d or v450g, whereas the third contains both mutations. analysis of the starch breakdown-profile of these three mutants, as well as of the wild-type, a ... | 2006 | 16197365 |
| identification of biogenic dimethyl selenodisulfide in the headspace gases above genetically modified escherichia coli. | escherichia coli jm109 cells were modified to express the genes encoded in a 3.8-kb chromosomal dna fragment from a metalloid-resistant thermophile, geobacillus stearothermophilus v. manual headspace extraction was used to collect the gases for gas chromatography with fluorine-induced sulfur chemiluminescence analysis while solid-phase microextraction was used for sample collection in gas chromatography/mass spectrometry (gc/ms) analysis. when grown in the presence of selenate or selenite, these ... | 2006 | 16289446 |
| molecular structure of a 9-mda icosahedral pyruvate dehydrogenase subcomplex containing the e2 and e3 enzymes using cryoelectron microscopy. | the pyruvate dehydrogenase multienzyme complexes are among the largest multifunctional catalytic machines in cells, catalyzing the production of acetyl coa from pyruvate. we have previously reported the molecular architecture of an 11-mda subcomplex comprising the 60-mer icosahedral dihydrolipoyl acetyltransferase (e2) decorated with 60 copies of the heterotetrameric (alpha(2)beta(2)) 153-kda pyruvate decarboxylase (e1) from bacillus stearothermophilus (milne, j. l. s., shi, d., rosenthal, p. b. ... | 2006 | 16308322 |
| application of artificial neural networks to describe the combined effect of ph and nacl on the heat resistance of bacillus stearothermophilus. | a model for prediction of bacterial spore inactivation was developed. the influence of temperature, ph and nacl on the heat resistance of bacillus stearothermophilus spores was described using low-complexity, black box models based on artificial neural networks. literature data were used to build and train the neural network, and new experimental data were used to evaluate it. the neural network models gave better predictions than the classical quadratic response surface model in all the experim ... | 2006 | 16216369 |
| sterilization of bacterial spores by using supercritical carbon dioxide and hydrogen peroxide. | it was hypothesized that supercritical carbon dioxide (sc-co(2)) treatment could serve as an alternative sterilization method at various temperatures (40-105 degrees c), co(2) pressures (200-680 atm), and treatment times (25 min to 6 h), and with or without the use of a passive additive (distilled water, dh(2)o) or an active additive (hydrogen peroxide, h(2)o(2)). while previous researchers have shown that sc-co(2) possesses antimicrobial properties, sterilization effectiveness has not been show ... | 2007 | 16838346 |
| distinct modes of recognition of the lipoyl domain as substrate by the e1 and e3 components of the pyruvate dehydrogenase multienzyme complex. | two-dimensional (15)n-heteronuclear single-quantum coherence (hsqc) nmr studies with a di-domain (lipoyl domain+ linker+ peripheral subunit-binding domain) of the dihydrolipoyl acetyltransferase (e2) component of the pyruvate dehydrogenase complex of bacillus stearothermophilus allowed a molecular comparison of the need for lipoic acid to be covalently attached to the lipoyl domain in order to undergo reductive acetylation by the pyruvate decarboxylase (e1) component, in contrast with the abilit ... | 2007 | 17157320 |
| the stability of engineered thermostable neutral proteases from bacillus stearothermophilus in organic solvents and detergents. | engineered extremely thermostable variants of the thermolysin-like protease from bacillus stearothermophilus possessing an introduced disulfide bond g8c/n60c (double mutant, dm) and six additional amino acid substitutions in the exposed loop region 56-69 (boilysin, bln) have been probed with respect to stability toward water-miscible organic solvents and detergents. the solvent concentrations where 50% of enzyme activity were irreversibly lost (c(50)) decreased in the order methanol > 2-propanol ... | 2007 | 17163509 |
| use of hydrogen peroxide vapor for deactivation of mycobacterium tuberculosis in a biological safety cabinet and a room. | mycobacterium tuberculosis is an important human pathogen that is routinely cultured in clinical and research laboratories. m. tuberculosis can contaminate surfaces and is highly resistant to disinfection. we investigated whether hydrogen peroxide vapor (hpv) is effective for the deactivation of m. tuberculosis on experimentally contaminated surfaces in a biological safety cabinet (bsc) and a room. biological indicators (bis) consisting of an approximately 3-log(10) inoculum of m. tuberculosis o ... | 2007 | 17166957 |
| novel biocatalysts based on s-layer self-assembly of geobacillus stearothermophilus nrs 2004/3a: a nanobiotechnological approach. | the crystalline cell-surface (s) layer sgse of geobacillus stearothermophilus nrs 2004/3a represents a natural protein self-assembly system with nanometer-scale periodicity that is evaluated as a combined carrier/patterning element for the conception of novel types of biocatalyst aiming at the controllable display of biocatalytic epitopes, storage stability, and reuse. the glucose-1-phosphate thymidylyltransferase rmla is used as a model enzyme and chimeric proteins are constructed by translatio ... | 2007 | 17786898 |
| production of l -alanine by metabolically engineered escherichia coli. | escherichia coli w was genetically engineered to produce l: -alanine as the primary fermentation product from sugars by replacing the native d: -lactate dehydrogenase of e. coli sz194 with alanine dehydrogenase from geobacillus stearothermophilus. as a result, the heterologous alanine dehydrogenase gene was integrated under the regulation of the native d: -lactate dehydrogenase (ldha) promoter. this homologous promoter is growth-regulated and provides high levels of expression during anaerobic f ... | 2007 | 17874321 |
| formaldehyde gas inactivation of bacillus anthracis, bacillus subtilis, and geobacillus stearothermophilus spores on indoor surface materials. | to evaluate the decontamination of bacillus anthracis, bacillus subtilis, and geobacillus stearothermophilus spores on indoor surface materials using formaldehyde gas. | 2007 | 17897215 |
| alanine production in an h+-atpase- and lactate dehydrogenase-defective mutant of escherichia coli expressing alanine dehydrogenase. | previously, we reported that pyruvate production was markedly improved in tbla-1, an h(+)-atpase-defective escherichia coli mutant derived from w1485lip2, a pyruvate-producing e. coli k-12 strain. tbla-1 produced more than 30 g/l pyruvate from 50 g/l glucose by jar fermentation, while w1485lip2 produced only 25 g/l pyruvate (yokota et al. in biosci biotechnol biochem 58:2164-2167, 1994b). in this study, we tested the ability of tbla-1 to produce alanine by fermentation. the alanine dehydrogenase ... | 2007 | 17583806 |
| functional analysis of the translation factor aif2/5b in the thermophilic archaeon sulfolobus solfataricus. | the protein if2/eif5b is one of the few translation initiation factors shared by all three primary domains of life (bacteria, archaea, eukarya). despite its phylogenetic conservation, the factor is known to present marked functional divergences in the bacteria and the eukarya. in this work, the function in translation of the archaeal homologue (aif2/5b) has been analysed in detail for the first time using a variety of in vitro assays. the results revealed that the protein is a ribosome-dependent ... | 2007 | 17608795 |
| substrate specificities of wild and mutated farnesyl diphosphate synthases from bacillus stearothermophilus with artificial substrates. | to determine the substrate specificities of wild and mutated types of farnesyl diphosphate (fpp) synthases from bacillus stearothermophilus, we examined the reactivities of 8-hydroxygeranyl diphosphate (hogpp) and 8-methoxygeranyl diphosphate (ch(3)ogpp) as allylic substrate homologs. the wild-type fpp synthase reaction of hogpp (and ch(3)ogpp) with isopentenyl diphosphate (ipp) gave hydroxyfarnesyl- (and methoxyfarnesyl-) diphosphates that stopped at the first stage of condensation. on the othe ... | 2007 | 17617711 |
| bacillus stearothermophilus pcra monomer is a single-stranded dna translocase but not a processive helicase in vitro. | structural studies of the bacillus stearothermophilus pcra protein along with biochemical studies of the single-stranded (ss) dna translocation activity of pcra monomers have led to the suggestion that a pcra monomer possesses processive helicase activity in vitro. yet definitive studies testing whether the pcra monomer possesses processive helicase activity have not been performed. here we show, using single turnover kinetic methods, that monomers of pcra are able to translocate along ssdna, in ... | 2007 | 17631491 |
| structure-specificity relationships of an intracellular xylanase from geobacillus stearothermophilus. | geobacillus stearothermophilus t-6 is a thermophilic gram-positive bacterium that produces two selective family 10 xylanases which both take part in the complete degradation and utilization of the xylan polymer. the two xylanases exhibit significantly different substrate specificities. while the extracellular xylanase (xt6; mw 43.8 kda) hydrolyzes the long and branched native xylan polymer, the intracellular xylanase (ixt6; mw 38.6 kda) preferentially hydrolyzes only short xylo-oligosaccharides. ... | 2007 | 17642511 |
| high-affinity interaction between the s-layer protein sbsc and the secondary cell wall polymer of geobacillus stearothermophilus atcc 12980 determined by surface plasmon resonance technology. | surface plasmon resonance studies using c-terminal truncation forms of the s-layer protein sbsc (recombinant sbsc consisting of amino acids 31 to 270 [rsbsc(31-270)] and rsbsc(31-443)) and the secondary cell wall polymer (scwp) isolated from geobacillus stearothermophilus atcc 12980 confirmed the exclusive responsibility of the n-terminal region comprising amino acids 31 to 270 for scwp binding. quantitative analyses indicated binding behavior demonstrating low, medium, and high affinities. | 2007 | 17644609 |
| structure determination and biochemical studies on bacillus stearothermophilus e53q serine hydroxymethyltransferase and its complexes provide insights on function and enzyme memory. | serine hydroxymethyltransferase (shmt) belongs to the alpha-family of pyridoxal 5'-phosphate-dependent enzymes and catalyzes the reversible conversion of l-ser and tetrahydrofolate to gly and 5,10-methylene tetrahydrofolate. 5,10-methylene tetrahydrofolate serves as a source of one-carbon fragment in many biological processes. shmt also catalyzes the tetrahydrofolate-independent conversion of l-allo-thr to gly and acetaldehyde. the crystal structure of bacillus stearothermophilus shmt (bsshmt) s ... | 2007 | 17651438 |
| geobacillus stearothermophilus lv cada gene mediates resistance to cadmium, lead and zinc in znta mutants of salmonella entérica serovar typhimurium. | salmonella entérica serovar typhimurium cells expressing the cada gene of geobacillus stearothermophilus lv exhibit a hypersensitive phenotype to cadmium chloride. deletion of the orf stm3576 from the salmonella genome resulted in cadmium, lead and zinc sensitivity, confirming that this orf is a homologue of the znta gene. the observed sensitivity was reverted upon expression of the g. stearothermophilus lv cada gene. these results indicate that the cada gene product is involved in cd, pb and zn ... | 2007 | 17657347 |
| crystal structure and ligand binding properties of the truncated hemoglobin from geobacillus stearothermophilus. | a novel truncated hemoglobin has been identified in the thermophilic bacterium geobacillus stearothermophilus (gs-trhb). the protein has been expressed in escherichia coli, the 3d crystal structure (at 1.5 angstroms resolution) and the ligand binding properties have been determined. the distal heme pocket displays an array of hydrogen bonding donors to the iron-bound ligands, including tyr-b10 on one side of the heme pocket and trp-g8 indole nitrogen on the opposite side. at variance with the hi ... | 2007 | 17126283 |
| characterization of a bifunctional aminoacylase/carboxypeptidase from radioresistant bacterium deinococcus radiodurans r1. | the gene encoding a deinococcus radiodurans r1 bifunctional aminoacylase/carboxypeptidase (dr_acy/cp) was amplified by polymerase chain reaction and cloned into pqe-30 to generate pqe-drac. the cloned gene consists of an open reading frame of 1197 bp encoding a protein with a molecular mass of 42,729 da. the predicted amino acid sequence shows high homology with those of geobacillus kaustophilus aminoacylase, geobacillus stearothermophilus aminoacylase, pyrococcus horikoshii carboxypeptidase/ami ... | 2007 | 17129628 |
| a two-component system regulates the expression of an abc transporter for xylo-oligosaccharides in geobacillus stearothermophilus. | geobacillus stearothermophilus t-6 utilizes an extensive and highly regulated hemicellulolytic system. the genes comprising the xylanolytic system are clustered in a 39.7-kb chromosomal segment. this segment contains a 6-kb transcriptional unit (xyndcefg) coding for a potential two-component system (xyndc) and an atp-binding cassette (abc) transport system (xynefg). the xynd promoter region contains a 16-bp inverted repeat resembling the operator site for the xylose repressor, xylr. xylr was fou ... | 2007 | 17142383 |
| a cold-active and thermostable alcohol dehydrogenase of a psychrotorelant from antarctic seawater, flavobacterium frigidimaris kuc-1. | an nad(+)-dependent alcohol dehydrogenase of a psychrotorelant from antarctic seawater, flavobacterium frigidimaris kuc-1 was purified to homogeneity with an overall yield of about 20% and characterized enzymologically. the enzyme has an apparent molecular weight of 160k and consists of four identical subunits with a molecular weight of 40k. the pi value of the enzyme and its optimum ph for the oxidation reaction were determined to be 6.7 and 7.0, respectively. the enzyme contains 2 gram-atoms z ... | 2007 | 17072683 |
| the smr gene resides on a novel plasmid psp187 identified in a staphylococcus pasteuri isolate recovered from unpasteurized milk. | this work describes a novel plasmid psp187 (5550 bp) carrying the small multidrug resistance determinant smr encoding resistance to quaternary ammonium compounds (qacs). psp187 was identified in a staphylococcus pasteuri isolate recovered from bulk milk in a dairy cattle herd in norway. sequence analysis revealed 6 putative orfs in addition to the smr gene within a cassette with identical genetic organization to that found in the psk41-like staphylococcus aureus plasmid ptz22. a protein homology ... | 2007 | 17074390 |
| crystal structure of tryptophanyl-trna synthetase complexed with adenosine-5' tetraphosphate: evidence for distributed use of catalytic binding energy in amino acid activation by class i aminoacyl-trna synthetases. | tryptophanyl-trna synthetase (trprs) is a functionally dimeric ligase, which specifically couples hydrolysis of atp to amp and pyrophosphate to the formation of an ester bond between tryptophan and the cognate trna. trprs from bacillus stearothermophilus binds the atp analogue, adenosine-5' tetraphosphate (aqp) competitively with atp during pyrophosphate exchange. estimates of binding affinity from this competitive inhibition and from isothermal titration calorimetry show that aqp binds 200 time ... | 2007 | 17428498 |
| species diversity and substrate utilization patterns of thermophilic bacterial communities in hot aerobic poultry and cattle manure composts. | this study investigated the species diversity and substrate utilization patterns of culturable thermophilic bacterial communities in hot aerobic poultry and cattle manure composts by coupling 16s rdna analysis with biolog data. based on the phylogenetic relationships of 16s rdna sequences, 34 thermophilic (grown at 60 degrees c) bacteria isolated during aerobic composting of poultry manure and cattle manure were classified as bacillus licheniformis, b. atrophaeus, geobacillus stearothermophilus, ... | 2007 | 17450396 |
| inxy and sexy, compact heterologous reporter proteins for mammalian cells. | mammalian reporter proteins are essential for gene-function analysis, drugscreening initiatives and as model product proteins for biopharmaceutical manufacturing. bacillus subtilis can maintain its metabolism by secreting xylanase a (xyna), which converts xylan into shorter xylose oligosaccharides. xyna is a family 11 xylanase monospecific for d-xylose containing substrates. mammalian cells transgenic for constitutive expression of wild-type xyna showed substantial secretion of this prokaryotic ... | 2007 | 17461419 |
| characterization and structural modeling of a new type of thermostable esterase from thermotoga maritima. | a bioinformatic screening of the genome of the hyperthermophilic bacterium thermotoga maritima for ester-hydrolyzing enzymes revealed a protein with typical esterase motifs, though annotated as a hypothetical protein. to confirm its putative esterase function the gene (estd) was cloned, functionally expressed in escherichia coli and purified to homogeneity. recombinant estd was found to exhibit significant esterase activity with a preference for short acyl chain esters (c4-c8). the monomeric enz ... | 2007 | 17466017 |
| a rapid and efficient method for cloning genes of type ii restriction-modification systems by use of a killer plasmid. | we present a method for cloning restriction-modification (r-m) systems that is based on the use of a lethal plasmid (pkiller). the plasmid carries a functional gene for a restriction endonuclease having the same dna specificity as the r-m system of interest. the first step is the standard preparation of a representative, plasmid-borne genomic library. then this library is transformed with the killer plasmid. the only surviving bacteria are those which carry the gene specifying a protective dna m ... | 2007 | 17468281 |
| molecular determinants of substrate recognition in thermostable alpha-glucosidases belonging to glycoside hydrolase family 13. | bacillus stearothermophilus alpha-1,4-glucosidase (bs) is highly specific for alpha-1,4-glucosidic bonds of maltose, maltooligosaccharides and alpha-glucans. bacillus thermoglucosdasius oligo-1,6-glucosidase (bt) can specifically hydrolyse alpha-1,6 bonds of isomaltose, isomaltooligosaccharides and alpha-limit dextrin. the two enzymes have high homology in primary structure and belong to glycoside hydrolase family 13, which contain four conservative regions (i, ii, iii and iv). the two enzymes a ... | 2007 | 17525102 |
| structure, flexibility, and mechanism of the bacillus stearothermophilus recu holliday junction resolvase. | here we report a high resolution structure of recu-holliday junction resolvase from bacillus stearothermophilus. the functional unit of recu is a homodimer that contains a "mushroom" like structure with a rigid cap and two highly flexible loops extending outwards. these loops appear to be highly flexible/dynamic, and presumably are directly involved in dna binding and holding it for catalysis. structural modifications of both the protein and dna upon their interaction are essential for catalysis ... | 2007 | 17557334 |
| dynamic allostery in the ring protein trap. | we have discovered distinct, characteristic differences in the thermodynamic signatures of tryptophan binding by trp rna-binding attenuation protein (trap) from two different bacterial species. the trap 11mer ring binds 11 molecules of tryptophan at symmetry-related sites. tryptophan binding to bacillus stearothermophilus trap is not cooperative, but isothermal titration calorimetry shows that filling the first tryptophan binding sites of bacillus subtilis trap has a marked effect on the thermod ... | 2007 | 17559872 |
| reengineering cca-adding enzymes to function as (u,g)- or dcdcda-adding enzymes or poly(c,a) and poly(u,g) polymerases. | cca-adding enzymes build and repair the 3'-terminal cca sequence of trna. these unusual rna polymerases use either a ribonucleoprotein template (class i) or pure protein template (class ii) to form mock base pairs with the watson-crick edges of incoming ctp and atp. guided by the class ii bacillus stearothermophilus cca-adding enzyme structure, we introduced mutations designed to reverse the polarity of hydrogen bonds between the nucleobases and protein template. we were able to transform the cc ... | 2007 | 17179213 |
| purification, crystallization and preliminary x-ray analysis of the bseci dna methyltransferase from bacillus stearothermophilus in complex with its cognate dna. | the dna methyltransferase m.bseci from bacillus stearothermophilus (ec 2.1.1.72), a 579-amino-acid enzyme, methylates the n6 atom of the 3' adenine in the sequence 5'-atcgat-3'. m.bseci was crystallized in complex with its cognate dna. the crystals were found to belong to the hexagonal space group p6, with unit-cell parameters a = b = 87.0, c = 156.1 a, beta = 120.0 degrees and one molecule in the asymmetric unit. two complete data sets were collected at wavelengths of 1.1 and 2.0 a to 2.5 and 2 ... | 2007 | 17183163 |
| the dtdp-4-dehydro-6-deoxyglucose reductase encoding fcd gene is part of the surface layer glycoprotein glycosylation gene cluster of geobacillus tepidamans gs5-97t. | the glycan chain of the s-layer protein of geobacillus tepidamans gs5-97(t) consists of disaccharide repeating units composed of l-rhamnose and d-fucose, the latter being a rare constituent of prokaryotic glycoconjugates. although biosynthesis of nucleotide-activated l-rhamnose is well established, d-fucose biosynthesis is less investigated. the conversion of alpha-d-glucose-1-phosphate into thymidine diphosphate (dtdp)-4-dehydro-6-deoxyglucose by the sequential action of rmla (glucose-1-phospha ... | 2007 | 17202151 |
| linamarase activities in bacillus spp. responsible for thermophilic aerobic digestion of agricultural wastes for animal nutrition. | thermophilic bacillus spp. isolated from thermophilic aerobic digestion (tad) of model agricultural slurry were screened for ability to secret linamarase activity and degrade linamarin, a cyanogenic glycoside toxin abundant in cassava. screening was performed by both linamarin - picrate assay and by p-nitrophenyl beta-d-glucoside (pnpg) degradation, and results of both assays were related. linamarase positive isolates were identified as bacillus coagulans, bacillus licheniformis and bacillus ste ... | 2007 | 17207614 |