| characterization of thermophilic bacteria using surface-enhanced raman scattering. | surface-enhanced raman scattering (sers) can provide molecular-level information about the molecules and molecular structures in the vicinity of nanostructured noble metal surfaces such as gold and silver. the three thermophilic bacteria bacillus licheniformis, geobacillus stearothermophilus, and geobacillus pallidus, a gram-negative bacterium e. coli, and a gram-positive bacterium b. megaterium are comparatively characterized using sers. the sers spectra of thermophilic bacteria are similar, wh ... | 2008 | 19007464 |
| isolation of the phe-operon from g. stearothermophilus comprising the phenol degradative meta-pathway genes and a novel transcriptional regulator. | geobacillus stearothermophilus is able to utilize phenol as a sole carbon source. a dna fragment encoding a phenol hydroxylase catalyzing the first step in the meta-pathway has been isolated previously. based on these findings a pcr-based dna walk was performed initially to isolate a catechol 2,3-dioxygenase for biosensoric applications but was continued to elucidate the organisation of the genes encoding the proteins for the metabolization of phenol. | 2008 | 19014555 |
| structural and functional studies of bacillus stearothermophilus serine hydroxymethyltransferase: the role of asn(341), tyr(60) and phe(351) in tetrahydrofolate binding. | shmt (serine hydoxymethyltransferase), a type i pyridoxal 5'-phosphate-dependent enzyme, catalyses the conversion of l-serine and thf (tetrahydrofolate) into glycine and 5,10-methylene thf. shmt also catalyses several thf-independent side reactions such as cleavage of beta-hydroxy amino acids, transamination, racemization and decarboxylation. in the present study, the residues asn(341), tyr(60) and phe(351), which are likely to influence thf binding, were mutated to alanine, alanine and glycine ... | 2009 | 19046138 |
| crystallization and preliminary crystallographic studies of the recombinant l-n-carbamoylase from geobacillus stearothermophilus cect43. | n-carbamoyl-l-amino-acid amidohydrolases (l-n-carbamoylases; ec 3.5.1.87) hydrolyze the carbon-nitrogen bond of the ureido group in n-carbamoyl-l-alpha-amino acids. these enzymes are commonly used in the production of optically pure natural and non-natural l-amino acids using the ;hydantoinase process'. recombinant l-n-carbamoylase from geobacillus stearothermophilus cect43 has been expressed, purified and crystallized by hanging-drop vapour diffusion. x-ray data were collected to a resolution o ... | 2008 | 19052368 |
| purification, crystallization and preliminary crystallographic analysis of wsaf, an essential rhamnosyltransferase from geobacillus stearothermophilus. | the beta1,2-rhamnosyltransferase wsaf is involved in the biosynthesis of a polyrhamnan chain which is attached to the surface-layer protein from geobacillus stearothermophilus nrs 2004/3a. the enzyme belongs to the large retaining gt4 family. to date, no structure of a rhamnosyltransferase has been published. recombinant purified native wsaf has been crystallized, resulting in crystals that belonged to space group p2(1)2(1)2(1) with unit-cell parameters a = 50.5, b = 56.1, c = 276.8 a and diffra ... | 2008 | 19052376 |
| comparison studies of the human heart and bacillus stearothermophilus lactate dehydrogreanse by transition path sampling. | transition path sampling is a well-known technique that generates reactive paths ensembles. due to the atomic detail of these reactive paths, information about chemical mechanisms can be obtained. we present here a comparative study of bacillus stearothermophilus and human heart homologues of lactate dehydrogenase (ldh). a comparison of the transition path ensemble of both enzymes revealed that small differences in the active site reverses the order of the particle transfer of the chemical step. ... | 2009 | 19053545 |
| identification of new secreted proteins and secretion of heterologous amylase by c. glutamicum. | in this study, secreted corynebacterium glutamicum proteins were investigated by two-dimensional gel electrophoresis. around 100 spots observed in the ph range 4.5-5.5 had molecular masses that varied from 10 to 50 kda. upon n-terminal amino acid sequence analysis by edman degradation, two of them were hits to two hypothetical proteins encoded by cgr_1176 and cgr_2070 on c. glutamicum r genome, respectively. active-form alpha-amylase derived from geobacillus stearothermophilus was successfully s ... | 2009 | 19066885 |
| snapshots of catalysis in the e1 subunit of the pyruvate dehydrogenase multienzyme complex. | the pyruvate dehydrogenase multienzyme assembly (pdh) generates acetyl coenzyme a and reducing equivalents from pyruvate in a multiple-step process that is a nexus of central metabolism. we report crystal structures of the geobacillus stearothermophilus pdh e1p subunit with ligands that mimic the prereaction complex and the postdecarboxylation product. the structures implicate residues that help to orient substrates, nurture intermediates, and organize surface loops so that they can engage a mob ... | 2008 | 19081062 |
| antioxidant activity of biotransformed sex hormones facilitated by bacillus stearothermophilus. | bacillus stearothermophilus, a thermophilic bacterium isolated from kuwaiti desert, when incubated with exogenous progesterone for 10 days at 65 degrees c produced two monohydroxylated, two dihydroxy isomers of progesterone and a b-seco compound. these metabolites were purified by tlc and hplc followed by their identification through (1)h, (13)c nmr and other spectroscopic data. microbial hydroxylation of 17beta-estradiol resulted in the production of estrone. the effect of some inducers resulte ... | 2008 | 19082955 |
| thermodynamic analysis reveals a temperature-dependent change in the catalytic mechanism of bacillus stearothermophilus tyrosyl-trna synthetase. | catalysis of trna(tyr) aminoacylation by tyrosyl-trna synthetase can be divided into two steps. in the first step, tyrosine is activated by atp to form the tyrosyl-adenylate intermediate. in the second step, the tyrosyl moiety is transferred to the 3' end of trna. to investigate the roles that enthalpic and entropic contributions play in catalysis by bacillus stearothermophilus tyrosyl-trna synthetase (tyrrs), the temperature dependence for the activation of tyrosine and subsequent transfer to t ... | 2009 | 19098308 |
| evaluation of a microbiological multi-residue system on the detection of antibacterial substances in ewe milk. | to protect both, public health and the dairy industry, from the presence of antibiotic residues in milk, control programmes have been established, which include the needed screening tests. this work focuses on the application of a microbiological multi-residue system in ewe milk, a method based on the use of six different plates, each seeded with one of the following bacteria: geobacillus stearothermophilus var. calidolactis (beta-lactams), bacillus subtilis at ph 8.0 (aminoglycosides), kocuria ... | 2009 | 19100896 |
| how to switch off a histidine kinase: crystal structure of geobacillus stearothermophilus kinb with the inhibitor sda. | entry to sporulation in bacilli is governed by a histidine kinase phosphorelay, a variation of the predominant signal transduction mechanism in prokaryotes. sda directly inhibits sporulation histidine kinases in response to dna damage and replication defects. we determined a 2.0-a-resolution x-ray crystal structure of the intact cytoplasmic catalytic core [comprising the dimerization and histidine phosphotransfer domain (dhp domain), connected to the atp binding catalytic domain] of the geobacil ... | 2009 | 19101565 |
| molecular dynamics study of the structure, flexibility and dynamics of thermostable l1 lipase at high temperatures. | molecular dynamics (md) simulations have been used to understand how protein structure, dynamics, and flexibility are affected by adaptation to high temperature for several years. we report here the results of the high temperature md simulations of bacillus stearothermophilus l1 (l1 lipase). we found that the n-terminal moiety of the enzyme showed a high flexibility and dynamics during high temperature simulations which preceded and followed by clear structural changes in two specific regions; t ... | 2009 | 19130194 |
| kinetics of deoxy-ctp incorporation opposite a dg-c8-n-2-aminofluorene adduct by a high-fidelity dna polymerase. | the model carcinogen n-2-acetylaminofluorene covalently binds to the c8 position of guanine to form two adducts, the n-(2'-deoxyguanosine-8-yl)-aminofluorene (g-af) and the n-2-(2'-deoxyguanosine-8-yl)-acetylaminofluorene (g-aaf). although they are chemically closely related, their biological effects are strongly different and they are processed by different damage tolerance pathways. g-af is bypassed by replicative and high-fidelity polymerases, while specialized polymerases ensure synthesis pa ... | 2009 | 19150355 |
| cloning, purification and characterization of geobacillus stearothermophilus v uroporphyrinogen-iii c-methyltransferase: evaluation of its role in resistance to potassium tellurite in escherichia coli. | the geobacillus stearothermophilus v coba gene encoding uroporphyrinogen-iii c-methyltransferase (also referred to as sumt) was cloned into escherichia coli and the recombinant enzyme was overexpressed and purified to homogeneity. the enzyme binds s-adenosyl-l-methionine and catalyzes the production of iii methyl uroporphyrinogen in vitro. e. coli cells expressing the g. stearothermophilus v coba gene exhibited increased resistance to potassium tellurite and potassium tellurate. site-directed mu ... | 2009 | 19154787 |
| immobilization of recombinant thermostable beta-galactosidase from bacillus stearothermophilus for lactose hydrolysis in milk. | a recombinant thermostable beta-galactosidase from bacillus stearothermophilus was immobilized onto chitosan using tris(hydroxymethyl)phosphine (thp) and glutaraldehyde, and a packed bed reactor was utilized to hydrolyze lactose in milk. the thermostability and enzyme activity of thp-immobilized beta-galactosidase during storage was superior to that of free and glutaraldehyde-immobilized enzymes. the thp-immobilized beta-galactosidase showed greater relative activity in the presence of ca(2+) th ... | 2009 | 19164659 |
| independent saturation of three trprs subsites generates a partially assembled state similar to those observed in molecular simulations. | two new crystal structures of bacillus stearothermophilus tryptophanyl-trna synthetase (trprs) afford evidence that a closed interdomain hinge angle requires a covalent bond between amp and an occupant of either pyrophosphate or tryptophan subsite. they also are within experimental error of a cluster of structures observed in a nonequilibrium molecular dynamics simulation showing partial active-site assembly. further, the highest energy structure in a minimum action pathway computed by using ela ... | 2009 | 19174517 |
| two crystal structures of lysyl-trna synthetase from bacillus stearothermophilus in complex with lysyladenylate-like compounds: insights into the irreversible formation of the enzyme-bound adenylate of l-lysine hydroxamate. | aminoacyl-trna synthetase forms an enzyme-bound intermediate, aminoacyladenylate in the amino-acid activation reaction. this reaction is monitored by measuring the atp-ppi exchange reason in which [(32)p]ppi is incorporated into atp. we previously reported that l-lysine hydroxamate completely inhibited the l-lysine-dependent atp-ppi exchange reaction catalysed by lysyl-trna synthetase from bacillus stearothermophilus (bslysrs). several experiments suggested that bslysrs can adenylate l-lysine hy ... | 2009 | 19174549 |
| rna and protein complexes of trp rna-binding attenuation protein characterized by mass spectrometry. | we have characterized both wild-type and mutant trap (trp rna-binding attenuation protein) from bacillus stearothermophilus , and their complexes with rna or its regulator anti-trap protein (at), by electrospray ionization mass spectrometry (esi-ms). wild-type trap mainly forms homo-11mer rings. the mutant used carries three copies of the trap monomer on a single polypeptide chain so that it associates to form a 12mer ring with four polypeptide molecules. mass spectra showed that both the wild-t ... | 2009 | 19219981 |
| bacterial contaminants in carbonated soft drinks sold in bangladesh markets. | a total of 225 carbonated soft drink (csd) samples from nine brands, from various locations in five metropolitan cities of bangladesh were examined to determine their bacteriological quality. most samples were not in compliance with microbiological standards set by organizations like the world health organization (who). pseudomonas aeruginosa was the predominant species with an incidence of 95%. streptococcus spp. and bacillus stearothermophilus were the next most prevalent with numbers ranging ... | 2009 | 19232446 |
| antifirming effects of starch degrading enzymes in bread crumb. | antifirming properties of amylases in bread crumb were evaluated in straight dough breadmaking and related to the amylolytically modified starch structure. amylase properties and action mechanisms determine starch structure in the breads and, hence, how amylopectin recrystallization, starch network formation, water redistribution, and water mobility occur during breadmaking and storage. a bacterial endo-alpha-amylase mainly hydrolyzed the longer starch polymer chains internally. it thus reduced ... | 2009 | 19239186 |
| scanning the corynebacterium glutamicum r genome for high-efficiency secretion signal sequences. | systematic screening of secretion proteins using an approach based on the completely sequenced genome of corynebacterium glutamicum r revealed 405 candidate signal peptides, 108 of which were able to heterologously secrete an active-form alpha-amylase derived from geobacillus stearothermophilus. these comprised 90 general secretory (sec)-type, 10 twin-arginine translocator (tat)-type and eight sec-type with presumptive lipobox peptides. only sec- and tat-type signals directed high-efficiency sec ... | 2009 | 19246745 |
| inactivation of geobacillus stearothermophilus in canned food and coconut milk samples by addition of enterocin as-48. | the cyclic bacteriocin enterocin as-48 was tested on a cocktail of two geobacillus stearothermophilus strains in canned food samples (corn and peas), and in coconut milk. as-48 (7 microg/g) reduced viable cell counts below detection levels in samples from canned corn and peas stored at 45 degrees c for 30 days. in coconut milk, bacterial inactivation by as-48 (1.75 microg/ml) was even faster. in all canned food and drink samples inoculated with intact g. stearothermophilus endospores, bacterioci ... | 2009 | 19269571 |
| rational design of bacillus stearothermophilus us100 l-arabinose isomerase: potential applications for d-tagatose production. | l-arabinose isomerases catalyze the bioconversion of d-galactose into d-tagatose. with the aim of producing an enzyme optimized for d-tagatose production, three bacillus stearothermophilus us100 l-arabinose isomerase mutants were constructed, purified and characterized. our results indicate that mutant q268k was significantly more acidotolerant and more stable at acidic ph than the wild-type enzyme. the n175h mutant has a broad optimal temperature range from 50 to 65 degrees c. with the aim of c ... | 2009 | 19278622 |
| construction of a gene knockout system for application in paenibacillus alvei ccm 2051t, exemplified by the s-layer glycan biosynthesis initiation enzyme wsfp. | the gram-positive bacterium paenibacillus alvei ccm 2051t is covered by an oblique surface layer (s-layer) composed of glycoprotein subunits. the s-layer o-glycan is a polymer of [-->3)-beta-d-galp-(1[alpha-d-glcp-(1-->6)]-->4)-beta-d-manpnac-(1-->] repeating units that is linked by an adaptor of -[groa-2-->opo2-->4-beta-d-manpnac-(1-->4)]-->3)-alpha-l-rhap-(1-->3)-alpha-l-rhap-(1-->3)-alpha-l-rhap-(1-->3)-beta-d-galp-(1--> to specific tyrosine residues of the s-layer protein. for elucidation of ... | 2009 | 19304819 |
| characterization of a novel thermostable carboxylesterase from geobacillus kaustophilus hta426 shows the existence of a new carboxylesterase family. | the gene gk3045 (741 bp) from geobacillus kaustophilus hta426 was cloned, sequenced, and overexpressed into escherichia coli rosetta (de3). the deduced protein was a 30-kda monomeric esterase with high homology to carboxylesterases from geobacillus thermoleovorans ny (99% identity) and geobacillus stearothermophilus (97% identity). this protein suffered a proteolytic cut in e. coli, and the problem was overcome by introducing a mutation in the gene (k212r) without affecting the activity. the res ... | 2009 | 19304850 |
| [purification of recombinant dna methyltransferase m2.bstse from nickase-modification system nm.bstsei and study of the enzyme properties]. | the operon of nickase-modification system from bacillus stearothermophilus se-589 (recognition site 5'-gagtc-3') includes two dna methyltransferase genes: bstseim1 and bstseim2. gene encoding dna methyltransferase m2.bstsei was cloned in pjw vector and expressed in e. coli cells. the enzyme m2.bstsei has been isolated by chromatographic purification. m2.bstsei displays maximum activity at 55 degrees c and ph 7.5. the enzyme modifies adenine in dna sequence 5'-gagtc-3' and has substrate specifici ... | 2009 | 19334521 |
| simultaneous treatment of washing, disinfection and sterilization using ultrasonic levitation, silver electrolysis and ozone oxidation. | a new type of ultrasonic washer-disinfector-sterilizer, able to clean, disinfect and sterilize most kinds of reusable medical devices, has been developed by using the ultrasonic levitation function with umbrella-shape oscillators and ozone bubbling together with sterilization carried out by silver electrolysis. we have examined the biomedical and physicochemical performance of this instrument. prokariotic and gram-negative escherichia coli and eukariotic saccharomyces cerevisiae were killed by s ... | 2009 | 19344093 |
| discrimination between right and wrong purine dntps by dna polymerase i from bacillus stearothermophilus. | we used a series of datp and dgtp analogues to determine how dna polymerase i from bacillus stearothermophilus (bf), a prototypical a family polymerase, uses n-1, n(2), n-3, and n(6) of purine dntps to differentiate between right and wrong nucleotide incorporation. altering any of these nitrogens had two effects. first, it decreased the efficiency of correct incorporation of the resulting dntp analogue, with the loss of n-1 and n-3 having the most severe effects. second, it dramatically increase ... | 2009 | 19348507 |
| interaction of if2 with the ribosomal gtpase-associated center during 70s initiation complex formation. | addition of an escherichia coli 50s subunit (50s(cy5)) containing a cy5-labeled l11 n-terminal domain (l11-ntd) within the gtpase-associated center (gac) to an e. coli 30s initiation complex (30sic(cy3)) containing cy3-labeled initiation factor 2 complexed with gtp leads to rapid development of a fret signal during formation of the 70s initiation complex (70sic). initiation factor 2 (if2) and elongation factor g (ef-g) induce similar changes in ribosome structure. here we show that such similari ... | 2009 | 19366171 |
| structural and mutational analyses of deinococcus radiodurans uvra2 provide insight into dna binding and damage recognition by uvras. | uvra proteins are key actors in dna damage repair and play an essential role in prokaryotic nucleotide excision repair (ner), a pathway that is unique in its ability to remove a broad spectrum of dna lesions. understanding the dna binding and damage recognition activities of the uvra family is a critical component for establishing the molecular basis of this process. here we report the structure of the class ii uvra2 from deinococcus radiodurans in two crystal forms. these structures, coupled wi ... | 2009 | 19368888 |
| [transglycosylation of benzo[h]quinazolines]. | this was the first study to apply cyclodextrin glucanotransferases (cgtase, ec 2.4.1.19) produced by mesophilic, thermophilic, alkaliphilic, and halophilic bacilli as well as pullulanase, beta-amylase, beta-galactosidase, and beta-fructofuranosidase for transglycosylation of benzo[h]quinazolines. the combination of cgtase produced by bacillus stearothermophilus st-88 and gamma-cyclodextrin (cd) used as a donor of glucosyl residues was the most efficient. the derivatives obtained are water-solubl ... | 2009 | 19382700 |
| geobacillus stearothermophilus 6-phosphogluconate dehydrogenase complexed with 6-phosphogluconate. | two crystal structures of recombinant geobacillus stearothermophilus 6-phosphogluconate dehydrogenase (gs6pdh) in complex with the substrate 6-phosphogluconate have been determined at medium resolution. gs6pdh shares significant sequence identity and structural similarity with the enzymes from lactococcus lactis, sheep liver and the protozoan parasite trypanosoma brucei, for which a range of structures have previously been reported. comparisons indicate that amino-acid sequence conservation is m ... | 2009 | 19407374 |
| robust experimental design for optimizing the microbial inhibitor test for penicillin detection in milk. | to use experimental design techniques and a multiple logistic regression model to optimize a microbiological inhibition test with dichotomous response for the detection of penicillin g in milk. | 2009 | 19413756 |
| allosteric regulation of the primase (dnag) activity by the clamp-loader (tau) in vitro. | during dna replication the helicase (dnab) recruits the primase (dnag) in the replisome to initiate the polymerization of new dna strands. dnab is attached to the tau subunit of the clamp-loader that loads the beta clamp and interconnects the core polymerases on the leading and lagging strands. the tau-dnab-dnag ternary complex is at the heart of the replisome and its function is likely to be modulated by a complex network of allosteric interactions. using a stable ternary complex comprising the ... | 2009 | 19415803 |
| models of single-molecule experiments with periodic perturbations reveal hidden dynamics in rna folding. | traditionally, microscopic fluctuations of molecules have been probed by measuring responses of an ensemble to perturbations. now, single-molecule experiments are capable of following fluctuations without introducing perturbations. however, dynamics not readily sampled at equilibrium should be accessible to nonequilibrium single-molecule measurements. in a recent study [qu, x. et al. proc. natl. acad. sci. u.s.a. 2008, 105, 6602-6607], the efficiency of fluorescence resonance energy transfer (fr ... | 2009 | 19415919 |
| the importance of an extra loop in the b-domain of an alpha-amylase from b. stearothermophilus us100. | to provide insight into the potential role of a loop in domain b of several bacterial alpha-amylases, molecular and structural investigation of bacillus stearothermophilus alpha-amylase (amy us100) was used as a model. combination deletion mutants of g(213), i(214) and g(215), described as a loop-forming on the surface bacterial amylases, were subjected to biochemical and structural investigation. thermoactivity, thermostability as well calcium requirement were studied for each mutant. thus, del ... | 2009 | 19422796 |
| effect of dimerization on the stability and catalytic activity of dihydrofolate reductase from the hyperthermophile thermotoga maritima. | in contrast to all other chromosomally encoded dihydrofolate reductases characterized so far, dihydrofolate reductase (dhfr) from the hyperthermophile thermotoga maritima forms a highly stable dimer. the dimer interface involves residues whose mobility is important for catalysis in monomeric dhfrs. here, we report the generation of a variant of dhfr from t. maritima, tmdhfr-v11d, in which a single amino acid replacement was sufficient to favor the monomeric form of the enzyme in the presence of ... | 2009 | 19453185 |
| the high-molecular-mass amylase (hmma) of geobacillus stearothermophilus atcc 12980 interacts with the cell wall components by virtue of three specific binding regions. | the complete nucleotide sequence encoding the high-molecular-mass amylase (hmma) of geobacillus stearothermophilus atcc 12980 was established by pcr techniques. based on the hmma gene sequence, the full-length rhmma, four n- or c-terminal rhmma truncations as well as three c-terminal rhmma fragments were cloned and heterologously expressed in escherichia coli. purified rhmma forms were used either for affinity studies with the recombinant (r) s-layer protein sbsc (rsbsc), peptidoglycan-containin ... | 2009 | 19460092 |
| structure of the sporulation histidine kinase inhibitor sda from bacillus subtilis and insights into its solution state. | the crystal structure of the dna-damage checkpoint inhibitor of sporulation, sda, from bacillus subtilis, has been solved by the mad technique using selenomethionine-substituted protein. the structure closely resembles that previously solved by nmr, as well as the structure of a homologue from geobacillus stearothermophilus solved in complex with the histidine kinase kinb. the structure contains three molecules in the asymmetric unit. the unusual trimeric arrangement, which lacks simple internal ... | 2009 | 19465772 |
| cold plasma technology: bactericidal effects on geobacillus stearothermophilus and bacillus cereus microorganisms. | cold plasma, also known as low temperature atmospheric pressure plasma (ltapp) is a novel technology consisting of neutral and charged particles, including free radicals, which can be used to destroy or inactivate microorganisms. research has been conducted regarding the effect of cold plasma on gram-positive bacteria; however, there is limited research regarding its ability to inactivate the spore-formers geobacillus stearothermophilus and bacillus cereus. | 2009 | 19470230 |
| safety evaluation of 1,4-alpha-glucan branching enzymes from bacillus stearothermophilus and aquifex aeolicus expressed in bacillus subtilis. | 1,4-alpha-glucan branching enzyme (be; ec 2.4.1.18) is a key biocatalyst in the synthesis of polysaccharides, and is therefore useful in the production of food ingredients. the bes evaluated in this study (be-01 and be-02) are obtained by fermentation of bacillus subtilis expressing the be gene from either bacillus stearothermophilus strain trbe14 or aquifex aeolicus strain vf5. the safety of be-01 and be-02 have not been previously evaluated, and therefore, both were subjected to standard toxic ... | 2009 | 19470400 |
| cloning and characterization of flagellin genes and identification of flagellin glycosylation from thermophilic bacillus species. | flagellin glycosylation was identified in bacillus sp. ps3 and geobacillus stearothermophilus. in vivo complementation showed that these flagellin genes did not restore the motility of a bacillus subtilis flagellin mutant, whereas the genes encoding non-glycosylated flagellin from geobacillus kaustophilus and bacillus sp. kps3 restored motility. moreover, four types of flagellins expressed in b. subtilis were not glycosylated. we speculate that glycosylation is required for flagellar filament as ... | 2009 | 19502747 |
| crystal structure of an inverting gh 43 1,5-alpha-l-arabinanase from geobacillus stearothermophilus complexed with its substrate. | arabinanases are glycosidases that hydrolyse alpha-(1-->5)- arabinofuranosidic linkages found in the backbone of the pectic polysaccharide arabinan. here we describe the biochemical characterization and the enzyme-substrate crystal structure of an inverting family 43 arabinanase from geobacillus stearothermophilus t-6 (abnb). based on viscosity and reducing power measurements, and based on product analysis for the hydrolysis of linear arabinan by abnb, the enzyme works in an endo mode of action. ... | 2009 | 19505290 |
| projection structure by single-particle electron microscopy of secondary transport proteins gltt, cits, and glts. | the structure of three secondary transporter proteins, gltt of bacillus stearothermophilus, cits of klebsiella pneumoniae, and glts of escherichia coli, was studied. the proteins were purified to homogeneity in detergent solution by ni(2+)-nta affinity chromatography, and the complexes were determined by bn-page to be trimeric, dimeric, and dimeric for gltt, cits, and glts, respectively. the subunit stoichiometry correlated with the binding affinity of the ni(2+)-nta resin for the protein comple ... | 2009 | 19518127 |
| toxicity assessment of the herbicide metolachlor comparative effects on bacterial and mitochondrial model systems. | metolachlor is one of the most intensively used chloroacetamide herbicides. however, its effects on the environment and on non-target animals and humans as well as its interference at a cell/molecular level have not yet been fully elucidated. the aim of this study was: firstly, to evaluate the potential toxicity of metolachlor at a cell/subcellular level by using two in vitro biological model systems (a strain of bacillus stearothermophilus and rat liver mitochondria); secondly, to evaluate the ... | 2009 | 19607910 |
| structural basis for thermostability revealed through the identification and characterization of a highly thermostable phosphotriesterase-like lactonase from geobacillus stearothermophilus. | a new enzyme homologous to phosphotriesterase was identified from the bacterium geobacillus stearothermophilus (gsp). this enzyme belongs to the amidohydrolase family and possesses the ability to hydrolyze both lactone and organophosphate (op) compounds, making it a phosphotriesterase-like lactonase (pll). gsp possesses higher op-degrading activity than recently characterized plls, and it is extremely thermostable. gsp is active up to 100 degrees c with an energy of activation of 8.0 kcal/mol to ... | 2009 | 19615330 |
| interaction of human dna polymerase alpha and dna polymerase i from bacillus stearothermophilus with hypoxanthine and 8-oxoguanine nucleotides. | to better understand how dna polymerases interact with mutagenic bases, we examined how human dna polymerase alpha (pol alpha), a b family enzyme, and dna polymerase from bacillus stearothermophilus (bf), an a family enzyme, generate adenine:hypoxanthine and adenine:8-oxo-7,8-dihydroguanine (8-oxog) base pairs. pol alpha strongly discriminated against polymerizing datp opposite 8-oxog, and removing n1, n(6), or n7 further inhibited incorporation, whereas removing n3 from datp dramatically increa ... | 2009 | 19642651 |
| safety evaluation of highly-branched cyclic dextrin and a 1,4-alpha-glucan branching enzyme from bacillus stearothermophilus. | highly-branched cyclic dextrin (hbcd), a dextrin food ingredient presently only used in japan, was investigated for digestibility and potential toxicity. hbcd was readily hydrolyzed in vitro to maltose and maltotriose by human salivary and porcine pancreatic alpha-amylases. incubation of hbcd with a rat intestinal homogenate, containing digestive enzymes, resulted in the formation of maltose, maltotriose, and maltotetraose, and with longer incubation times, resulted in the formation of glucose. ... | 2009 | 19651182 |
| evolved beta-galactosidases from geobacillus stearothermophilus with improved transgalactosylation yield for galacto-oligosaccharide production. | a mutagenesis approach was applied to the beta-galactosidase bgab from geobacillus stearothermophilus kve39 in order to improve its enzymatic transglycosylation of lactose into oligosaccharides. a simple screening strategy, which was based on the reduction of the hydrolysis of a potential transglycosylation product (lactosucrose), provided mutant enzymes possessing improved synthetic properties for the autocondensation product from nitrophenyl-galactoside and galacto-oligosaccharides (gos) from ... | 2009 | 19666723 |
| afnor validation of premi test, a microbiological-based screening tube-test for the detection of antimicrobial residues in animal muscle tissue. | premi test contains viable spores of a strain of bacillus stearothermophilus which is sensitive to antimicrobial residues, such as beta-lactams, tetracyclines, macrolides and sulphonamides. the growth of the strain is inhibited by the presence of antimicrobial residues in muscle tissue samples. premi test was validated according to afnor rules (french association for normalisation). the afnor validation was based on the comparison of reference methods (french official method, i.e. four plate tes ... | 2008 | 19680855 |
| evaluation of three different microbial inhibition tests for the detection of sulphamethazine residues in the edible tissues of rabbit. | the aim of the present study was to evaluate three microbial inhibition tests (mit) based on inhibition of growth of the test organisms: (a) four plate test (fpt) containing bacillus subtilis bga, (b) screening test for antibiotic residues (star) containing bacillus stearothermophilus var. calidolactis_atcc 10149 and (c) the premi(r)test containing bacillus stearothermophilus var. calidolactis. the tests were used to determine sulphamethazine (smz) residues in edible tissues of rabbit after oral ... | 2009 | 19680973 |
| molecular characterization of the alkb gene in the thermophilic geobacillus sp. strain mh-1. | an extremely thermophilic alkane-degrading bacterium, strain mh-1, was isolated from the deep subterranean petroleum reservoir in shengli oil field, pr china. based on its physiological characteristics and analysis of its 16s rrna gene sequence, strain mh-1 was identified as geobacillus stearothermophilus. strain mh-1 was able to grow at temperatures ranging from 50 to 72 degrees c and effectively degraded hexadecane as the sole carbon source at 70 degrees c. strain mh-1 degraded alkanes with di ... | 2009 | 19733653 |
| epr and endor characterization of the reactive intermediates in the generation of no by cryoreduced oxy-nitric oxide synthase from geobacillus stearothermophilus. | cryoreduction epr/endor/step-annealing measurements with substrate complexes of oxy-gsnos (3; gsnos is nitric oxide synthase from geobacillus stearothermophilus) confirm that compound i (6) is the reactive heme species that carries out the gsnos-catalyzed (stage i) oxidation of l-arginine to n-hydroxy-l-arginine (noha), whereas the active species in the (stage ii) oxidation of noha to citrulline and hno/no(-) is the hydroperoxy-ferric form (5). when 3 is reduced by tetrahydrobiopterin (bh4), ins ... | 2009 | 19754116 |
| the unstructured c-terminal extension of uvrd interacts with uvrb, but is dispensable for nucleotide excision repair. | during nucleotide excision repair (ner) in bacteria the uvrc nuclease and the short oligonucleotide that contains the dna lesion are removed from the post-incision complex by uvrd, a superfamily 1a helicase. helicases are frequently regulated by interactions with partner proteins, and immunoprecipitation experiments have previously indicated that uvrd interacts with uvrb, a component of the post-incision complex. we examined this interaction using 2-hybrid analysis and surface plasmon resonance ... | 2009 | 19762288 |
| crystal structures of leishmania mexicana phosphoglycerate mutase suggest a one-metal mechanism and a new enzyme subclass. | the structures of leishmania mexicana cofactor-independent phosphoglycerate mutase (lm ipgam) crystallised with the substrate 3-phosphoglycerate at high and low cobalt concentrations have been solved at 2.00- and 1.90-a resolutions. both structures are very similar and the active site contains both 3-phosphoglycerate and 2-phosphoglycerate at equal occupancies (50%). lm ipgam co-crystallised with the product 2-phosphoglycerate yields the same structure. two co(2+) are coordinated within the acti ... | 2009 | 19781556 |
| the n-terminal region is crucial for the thermostability of the g-domain of bacillus stearothermophilus ef-tu. | bacterial elongation factor tu (ef-tu) is a model monomeric g protein composed of three covalently linked domains. previously, we evaluated the contributions of individual domains to the thermostability of ef-tu from the thermophilic bacterium bacillus stearothermophilus. we showed that domain 1 (g-domain) sets up the basal level of thermostability for the whole protein. here we chose to locate the thermostability determinants distinguishing the thermophilic domain 1 from a mesophilic domain 1. ... | 2010 | 19800034 |
| insight into the stereospecificity of short-chain thermus thermophilus alcohol dehydrogenase showing pro-s hydride transfer and prelog enantioselectivity. | the stereochemistry of the hydride transfer in reactions catalyzed by nad(h)-dependent alcohol dehydrogenase from thermus thermophilus hb27 was determined by means of (1)h-nmr spectroscopy. the enzyme transfers the pro-s hydrogen of [4r-(2)h]nadh and exhibits prelog specificity. enzyme-substrate docking calculations provided structural details about the enantioselectivity of this thermophilic enzyme. these results give additional insights into the diverse active site architectures of the largely ... | 2010 | 19807673 |
| use of a new catabolite repression resistant promoter isolated from bacillus subtilis kcc103 for hyper-production of recombinant enzymes. | bacillus subtilis kcc103 hyper-produces alpha-amylase and the synthesis is resistant to carbon catabolite repression. the strain efficiently produced alpha-amylase in low cost agro-biomass based medium rich in simple sugars without catabolite repression. here, the catabolite repression resistant promoter (amyr4) of alpha-amylase was isolated from kcc103 and used to synthesize recombinant enzymes in b. subtilis. when the bgab gene encoding beta-galactosidase of bacillus stearothermophilus was clo ... | 2010 | 19815075 |
| aspergillusol a, an alpha-glucosidase inhibitor from the marine-derived fungus aspergillus aculeatus. | a new tyrosine-derived metabolite, aspergillusol a (4), was isolated on a gram scale, together with a methyl ester of 4-hydroxyphenylpyruvic acid oxime (5) and secalonic acid a, from the marine-derived fungus aspergillus aculeatus cri323-04. the tetraol in 4 was identified as erythritol by comparison of the 1h nmr spectrum of its benzoylated derivative with those of benzoylated erythritol (7) and d-threitol (8), as well as by cellulose-based chiral hplc analysis. aspergillusol a (4) selectively ... | 2009 | 19824618 |
| identifying assembly-inhibiting and assembly-tolerant sites in the sbsb s-layer protein from geobacillus stearothermophilus. | surface layer (s-layer) proteins self-assemble into two-dimensional crystalline lattices that cover the cell wall of all archaea and many bacteria. we have generated assembly-negative protein variants of high solubility that will facilitate high-resolution structure determination. assembly-negative versions of the s-layer protein sbsb from geobacillus stearothermophilus pv72/p2 were obtained using an insertion mutagenesis screen. the haemagglutinin epitope tag was inserted at 23 amino acid posit ... | 2010 | 19836402 |
| atomic substitution reveals the structural basis for substrate adenine recognition and removal by adenine dna glycosylase. | adenine dna glycosylase catalyzes the glycolytic removal of adenine from the promutagenic a.oxog base pair in dna. the general features of dna recognition by an adenine dna glycosylase, bacillus stearothermophilus muty, have previously been revealed via the x-ray structure of a catalytically inactive mutant protein bound to an a:oxog-containing dna duplex. although the structure revealed the substrate adenine to be, as expected, extruded from the dna helix and inserted into an extrahelical activ ... | 2009 | 19841264 |
| towards the structure of the c-terminal part of the s-layer protein sbsc. | the s-layer protein sbsc from geobacillus stearothermophilus atcc 12980 is the most prevalent single protein produced by the bacterium and covers the complete bacterial surface in the form of a two-dimensional crystalline monolayer. in order to elucidate the structural features of the assembly domains, several n-terminally truncated fragments of sbsc have been crystallized. crystals obtained from recombinant fragments showed anisotropic diffraction to a maximum of 3.5 a resolution using synchrot ... | 2009 | 19851018 |
| disentangling the web of allosteric communication in a homotetramer: heterotropic inhibition in phosphofructokinase from escherichia coli. | this study quantifies the contribution of each of the four unique inhibiting heterotropic interactions between the allosteric inhibitor, phosphoenolpyruvate (pep), and the substrate, fructose 6-phosphate (fru-6-p), in phosphofructokinase from escherichia coli (ecpfk). the unique heterotropic interactions, previously labeled by the distances between ligand binding sites, were isolated independently by constructing hybrid tetramers. of the four unique heterotropic pep-fru-6-p interactions, the 45 ... | 2009 | 19905012 |
| a new xylanase from thermoacidophilic alicyclobacillus sp. a4 with broad-range ph activity and ph stability. | we have identified a highly ph-adaptable and stable xylanase (xyna4) from the thermoacidophilic alicyclobacillus sp. a4, a strain that was isolated from a hot spring in yunnan province, china. the gene (xyna4) that encodes this xylanase was cloned, sequenced, and expressed in escherichia coli. it encodes a 338-residue polypeptide with a calculated molecular mass of 42.5 kda. the deduced amino acid sequence is most similar to (53% identity) an endo-1,4-beta-xylanase from geobacillus stearothermop ... | 2010 | 19916085 |
| ribosomal interaction of bacillus stearothermophilus translation initiation factor if2: characterization of the active sites. | infb-encoded translation initiation factor if2 contains a non-conserved n-terminal domain and two conserved domains (g and c) constituted by three (g1, g2 and g3) and two (c1 and c2) sub-domains. here, we show that: (i) bacillus stearothermophilus if2 complements in vivo an escherichia coli infb null mutation and (ii) the n-domain of b. stearothermophilus if2, like that of e. coli if2, provides a strong yet dispensable interaction with 30 s and 50 s subunits in spite of the lack of any size, seq ... | 2010 | 19917289 |
| endo- and exo-inulinases: enzyme-substrate interaction and rational immobilization. | three-dimensional models of exoinulinase from bacillus stearothermophilus and endoinulinase from aspergillus niger were built up by means of homology modeling. the crystal structure of exoinulinase from aspergillus awamori was used as a template, which is the sole structure of inulinase resolved so far. docking and molecular dynamics simulations were performed to investigate the differences between the two inulinases in terms of substrate selectivity. the analysis of the structural differences b ... | 2010 | 19941325 |
| absorption, steady-state fluorescence, fluorescence lifetime, and 2d self-assembly properties of engineered fluorescent s-layer fusion proteins of geobacillus stearothermophilus nrs 2004/3a. | s-layer fusion protein technology was used to design four different fluorescent fusion proteins with three different gfp mutants and the red fluorescent protein mrfp1. their absorption spectra, steady-state fluorescence, and fluorescence lifetime were investigated as a function of ph. it was found that fluorescence intensities and lifetime of the gfp mutant s-layer fusion proteins decreased about 50% between ph 6 and ph 5. the spectral properties of the red s-layer fusion protein were minimally ... | 2010 | 19954211 |
| on the importance of the small domain in the thermostability of thermoalkalophilic lipases from l1 and t1: insights from molecular dynamics simulation. | an all-atom level md simulation in explicit solvent at high temperature is a powerful technique to increase our knowledge about the structurally important regions modulating thermal stability in thermenzymes. in this respect, two large-sized thermoalkalophilic enzymes from bacillus stearothermophilus l1 (l1 lipase) and geobacillus zalihae strain t1 (t1 lipase) are well-established representatives. in this paper, comparative results from temperature-induced md simulations of both model systems at ... | 2010 | 19958281 |
| hydrolysis of amylopectin by amylolytic enzymes: structural analysis of the residual amylopectin population. | amylopectin fine structures were studied following limited hydrolysis of gelatinised waxy maize starch by amylases with a different level of inner chain attack (lica). this was done by size exclusion chromatography as well as by debranching the (partially hydrolysed) amylopectin samples and studying the size distributions of the released chains. alpha-amylases from bacillus amyloliquefaciens and aspergillus oryzae, with a relatively high lica, drastically altered amylopectin chain length distrib ... | 2010 | 19962130 |
| hydrolysis of amylopectin by amylolytic enzymes: level of inner chain attack as an important analytical differentiation criterion. | differences in amylase action pattern on amylopectin were demonstrated by the relation between the decrease in potassium iodide-iodine binding of waxy maize starch and the increase in reducing value during hydrolysis, as expressed by the rv(80) value (i.e., the reducing value for a potassium iodide-iodine binding value of 80% of that of the starting material). in the initial stages of the hydrolysis, the ratio of the increase in the level of reducing polysaccharides to the increase in the total ... | 2010 | 19962134 |
| a group i self-splicing intron in the flagellin gene of the thermophilic bacterium geobacillus stearothermophilus. | a group i intron that can be spliced in vivo and in vitro was identified in the flagellin gene of the thermophilic bacterium geobacillus stearothermophilus. we also found one or two intervening sequences (ivs) of flagellin genes in five additional bacterial species. furthermore, we report the presence of these sequences in two sites of a highly conserved region in the flagellin gene. | 2009 | 19966479 |
| characterization of thermostable fmn-dependent nadh azoreductase from the moderate thermophile geobacillus stearothermophilus. | the gene encoding an fmn-dependent nadh azoreductase, azrg, from thermophilic geobacillus stearothermophilus was cloned and functionally expressed in recombinant escherichia coli. purified recombinant azrg is a homodimer of 23 kda and bore fmn as a flavin cofactor. the optimal temperature of azrg was 85 degrees c for the degradation of methyl red (mr). azrg remained active for 1 h at 65 degrees c and for 1 month at 30 degrees c, demonstrating both superior thermostability and long-term stability ... | 2010 | 19997911 |
| dry thermal resistance of bacillus anthracis (sterne) spores and spores of other bacillus species: implications for biological agent destruction via waste incineration. | to obtain needed data on the dry thermal resistance of bacillus anthracis spores and other bacillus species for waste incinerator applications. | 2010 | 20015207 |
| antibacterial and antifungal studies of macrocyclic complexes of trivalent transition metal ions with their spectroscopic approach. | a new series of complexes of the type [m(c24h16n4)x]x2, where m = cr(iii), fe(iii), and mn(iii), x = cl-, no3-, and ch3coo-, has been synthesized by template condensation of 1,8-diaminonaphthalene and glyoxal in the presence of trivalent metal salts in methanolic medium. the complexes have been characterized with the help of elemental analysis, conductance measurements, magnetic measurements, and electronic, nmr, ir, and mass spectral studies. on the basis of these studies, a five-coordinate squ ... | 2010 | 20030506 |
| heterogeneous properties of individual molecules of beta-galactosidase from the thermophilic bacteria geobacillus stearothermophilus. | single enzyme molecule assays were performed on beta-galactosidase from the thermophilic bacteria geobacillus stearothermophilus using a capillary electrophoresis-based continuous flow assay and the substrate ddao-beta-d: -galactoside. the enzyme was found to be heterogeneous with respect to catalytic rate, electrophoretic mobility and activation energy of catalysis. catalytic rate was also found to vary over time for individual molecules at elevated temperature. comparison with beta-galactosida ... | 2010 | 20049517 |
| antioxidant activity of biotransformed sex hormones facilitated by bacillus stearothermophilus. | bacillus stearothermophilus, a thermophilic bacterium isolated from kuwaiti desert, when incubated with exogenous progesterone for 10 days at 65 degrees c produced two monohydroxylated, two dihydroxy isomers of progesterone and a b-seco compound. these metabolites were purified by tlc and hplc followed by their identification through (1)h, (13)c nmr and other spectroscopic data. microbial hydroxylation of 17beta-estradiol resulted in the production of estrone. the effect of some inducers resulte ... | 2010 | 20072930 |
| geobacillus sp., a thermophilic soil bacterium producing volatile antibiotics. | geobacillus, a bacterial genus, is represented by over 25 species of gram-positive isolates from various man-made and natural thermophilic areas around the world. an isolate of this genus (m-7) has been acquired from a thermal area near yellowstone national park, mt and partially characterized. the cells of this organism are globose (ca. 0.5 mu diameter), and they are covered in a matrix capsule which gives rise to elongate multicelled bacilliform structures (ranging from 3 to 12 mum) as seen by ... | 2010 | 20091406 |
| refolding of the non-specific neutral protease from bacillus stearothermophilus proceeds via an autoproteolytically sensitive intermediate. | a very thermostable variant of the thermolysin-like protease from bacillus stearothermophilus (g8c/n60c) was previously created by introduction of a disulfide bond into the cysteine-free pseudo-wild type variant (pwt) and thus fixing the unfolding region 56-69. in the present paper, we show that g8c/n60c and pwt can be reactivated from the completely unfolded states, accessible at >or=7.5m guanidine hydrochloride, and analyze the kinetics of folding, autoproteolytic degradation and aggregation. ... | 2010 | 20096501 |
| structural basis of substrate binding in wsaf, a rhamnosyltransferase from geobacillus stearothermophilus. | carbohydrate polymers are medically and industrially important. the s-layer of many gram-positive organisms comprises protein and carbohydrate polymers and forms an almost paracrystalline array on the cell surface. not only is this array important for the bacteria but it has potential application in the manufacture of commercially important polysaccharides and glycoconjugates as well. the s-layer glycoprotein glycan from geobacillus stearothermophilus nrs 2004/3a is mainly composed of repeating ... | 2010 | 20097205 |
| a mas nmr study of the bacterial abc transporter artmp. | atp-binding cassette (abc) transport systems facilitate the translocation of substances, like amino acids, across cell membranes energised by atp hydrolysis. this work describes first structural studies on the abc transporter artmp from geobacillus stearothermophilus in native lipid environment by magic-angle spinning nmr spectroscopy. the 2d crystals of artmp and 3d crystals of isolated artp were prepared in different nucleotide-bound or -unbound states. from selectively (13)c,(15)n-labelled ar ... | 2010 | 20099290 |
| magnetic circular dichroism spectroscopic characterization of the nos-like protein from geobacillus stearothermophilus (gsnos). | nitric oxide synthase (nos) catalyzes the nadph- and o(2)-dependent oxidation of l-arginine (l-arg) to nitric oxide (no) and citrulline via an n(g)-hydroxy-l-arginine (nha) intermediate. mammalian noss have been studied quite extensively; other eukaryotes and some prokaryotes appear to express nos-like proteins comparable to the oxygenase domain of mammalian noss. in this study, a recombinant nos-like protein from the thermostable bacterium geobacillus stearothermophilus (gsnos) has been charact ... | 2010 | 20110129 |
| alkaline ph-dependent differential unfolding characteristics of mesophilic and thermophilic homologs of dimeric serine hydroxymethyltransferase. | environmental variables such as ph can significantly influence the folding and stability of a protein molecule. in the present investigation, we compared the alkaline ph-induced unfolding of two homologous serine hydroxymethyltransferase from mesophilic bacillus subtilis (bsshmt) and thermophilic bacillus stearothermophilus (bstshmt) using various biophysical techniques. the thermophilic enzyme bstshmt was found to be more resistant to alkaline denaturation compared to its mesophilic counterpart ... | 2010 | 20152942 |
| the second extracellular loop of pore-forming subunits of atp-binding cassette transporters for basic amino acids plays a crucial role in interaction with the cognate solute binding protein(s). | in the thermophile geobacillus stearothermophilus, the uptake of basic amino acids is mediated by an abc transporter composed of the substrate binding protein (receptor) artj and a homodimer each of the pore-forming subunit, artm, and the nucleotide-binding subunit, artp. we recently identified two putative binding sites in artj that might interact with the art(mp)(2) complex, thereby initiating the transport cycle (a. vahedi-faridi et al., j. mol. biol. 375:448-459, 2008). here we investigated ... | 2010 | 20154136 |
| antibacterial activities of the extracts and conessine from holarrhena floribunda g. don. (apocynaceae). | the methanolic extract and conessine isolated from the stem bark of holarrhena floribunda (hf) were tested for their antibacterial activities on bacillus: bacillus cereus, bacillus subtilis, bacillus megaterium and bacillus stearothermophilus using the disc diffusion method. phytochemical analysis of the crude extract and fractions was also conducted. the inhibition parameters of the crude methanol extract and the total alkaloid fraction were determined using the macrodilution method. the result ... | 2007 | 20161899 |
| the structure of dinb from geobacillus stearothermophilus: a representative of a unique four-helix-bundle superfamily. | the crystal structure of the dinb gene product from geobacillus stearothermophilus (gsdinb) is reported at 2.5 a resolution. the dinb gene is one of the dna-damage-induced genes and the corresponding protein, dinb, is the founding member of a pfam family with no known function. the protein contains a four-helix up-down-down-up bundle that has previously been described in the literature in three disparate proteins: the enzyme mdmpi (mycothiol-dependent maleylpyruvate isomerase), yfit and ttha0303 ... | 2010 | 20208147 |
| [enzymatic transglycosylation of natural and modified nucleosides by immobilized thermostable nucleoside phosphorylases from geobacillus stearothermophilus]. | natural and modified purine nucleosides have been synthesized using the recombinant thermostable enzymes purine nucleoside phosphorylase ii (ec 2.4.2.1) and pyrimidine nucleoside phosphorylase (ec 2.4.2.2) from geobacillus stearothermophilus b-2194. the enzymes were produced in recombinant e. coli strains and covalently immobilized on aminopropylsilochrom ap-cpg-170 after heating the cell lysates and the removal of coagulated thermolabile proteins. the resulting preparations of thermostable nucl ... | 2009 | 20208582 |
| studying subunit-subunit interactions in a bacterial abc transporterby in vitro assembly. | the thermostable arginine abc transporter of geobacillus stearothermophilus consists of a solute binding protein, artj; a transmembrane subunit, artm; and a nucleotide-binding subunit, artp. an artm/his(6)-artp complex was functionally assembled from separately purified subunits as demonstrated by assaying stimulation of its atpase activity by arginine-loaded artj in proteoliposomes. studying in vitro assembly with variants carrying mutations in the conserved q loop and/or the eaa loop of artp a ... | 2010 | 20226162 |
| improved yields of cyclic nigerosylnigerose from starch by pretreatment with a thermostable branching enzyme. | cyclic nigerosylnigerose (cnn) is produced enzymatically from starch by the combined action of 6-alpha-glucosyltransferase and 3-alpha-isomaltosyltransferase. in our previous study, alpha-1,6-branching chains found in the structure of amylopectin and glycogen were shown to be favorable for cnn formation by the two enzymes. therefore, we examined whether the introduction of alpha-1,6-branch points into starch using the action of branching enzyme (be) could improve the yield of cnn from starch. th ... | 2010 | 20226381 |
| substrate specificity and biochemical properties of m3.bstf5i dna methyltransferase from the bstf5i restriction-modification system. | optimal conditions for dna methylation by the m3.bstf5i enzyme from bacillus stearothermophilus and kinetic parameters of lambda phage dna modification and that of a number of oligonucleotide substrates are established. comparison of m1.bstf5i and m3.bstf5i kinetic parameters revealed that with similar temperature optima and affinity for dna, m3.bstf5i has nearly fourfold lower turnover number (0.24 min(-1)) and modifies the hemimethylated recognition site with lower efficiency under optimal con ... | 2010 | 20331425 |
| structural study of carboxylesterase from hyperthermophilic bacteria geobacillus stearothermophilus by molecular dynamics simulation. | carboxylesterases are ubiquitous enzymes with important physiological, industrial and medical applications such as synthesis and hydrolysis of stereo specific compounds, including the metabolic processing of drugs, and antimicrobial agents. here, we have performed molecular dynamics simulations of carboxylesterase from hyperthermophilic bacterium geobacillus stearothermophilus (gsest) for 10ns each at five different temperatures namely at 300k, 343k, 373k, 473k and 500k. profiles of root mean sq ... | 2010 | 20347362 |
| preparation of carboxylated magnetic particles for the efficient immobilization of c-terminally lysine-tagged bacillus stearothermophilus aminopeptidase ii. | this article reports the synthesis and use of surface-modified iron oxide particles for the simultaneous purification and immobilization of bacillus stearothermophilus aminopeptidase ii (bsapii) tagged c-terminally with either tri- or nona-lysines (bsapii-lys(3/9)). the carboxylated magnetic particles were prepared by the simple co-precipitation of fe(3+)/fe(2+) in aqueous medium and then subsequently modified with adipic acid. transmission electron microscopy (tem) micrographs showed that the c ... | 2010 | 20373125 |
| enzymatic synthesis of glycosylated puerarin using maltogenic amylase from bacillus stearothermophilus expressed in bacillus subtilis. | the maltogenic amylase from bacillus stearothermophilus (bsma) is a valuable biocatalyst that has been used to transglycosylate natural glycosides to improve solubility. to ensure safety, bsma was produced in bacillus subtilis, using new shuttle vector-based expression vectors. the transglycosylation of puerarin was also conducted with crude bsma and analyzed. | 2010 | 20393999 |
| pcr detection of thermophilic spore-forming bacteria involved in canned food spoilage. | thermophilic bacteria that form highly heat-resistant spores constitute an important group of spoilage bacteria of low-acid canned food. a pcr assay was developed in order to rapidly trace these bacteria. three pcr primer pairs were designed from rrna gene sequences. these primers were evaluated for the specificity and the sensitivity of detection. two primer pairs allowed detection at the species level of geobacillus stearothermophilus and moorella thermoacetica/thermoautrophica. the other pair ... | 2010 | 20397018 |
| enhanced production of lipase by the thermophilic geobacillus stearothermophilus strain-5 using statistical experimental designs. | statistically based experimental designs were applied to optimize the cultural conditions for the production of a glycerol-inducible lipase from the thermophilic geobacillus stearothermophilus strain-5. the effect of nineteen culture conditions on enzyme production was evaluated using plackett-burman factorial design. tween 80, k(2)hpo(4), glycerol and glucose were the most significant factors in improving enzyme production. the selected parameters were then further investigated using central co ... | 2010 | 20412872 |
| a selection that reports on protein-protein interactions within a thermophilic bacterium. | many proteins can be split into fragments that exhibit enhanced function upon fusion to interacting proteins. while this strategy has been widely used to create protein-fragment complementation assays (pcas) for discovering protein-protein interactions within mesophilic organisms, similar assays have not yet been developed for studying natural and engineered protein complexes at the temperatures where thermophilic microbes grow. we describe the development of a selection for protein-protein inte ... | 2010 | 20418388 |
| detection of fusarium graminearum dna using a loop-mediated isothermal amplification (lamp) assay. | loop-mediated isothermal amplification (lamp) of dna is a simple, cost effective, and rapid method for the specific detection of genomic dna using a set of six oligonucleotide primers with eight binding sites hybridizing specifically to different regions of a target gene, and a thermophilic dna polymerase from geobacillus stearothermophilus for dna amplification. the method has been applied in various assays for the diagnosis of bacterial and viral infections of humans and animals, sexing of bov ... | 2010 | 20442002 |
| physicochemical characteristics of a thermostable gellan lyase from geobacillus stearothermophilus 98. | a purified thermostable gellan lyase, produced by a thermophilic bacterium, geobacillus stearothermophilus 98, was characterized in relation to its physicochemical properties. the gellan lyase was established to have a molecular weight of 216 kda, defined by capillary gel electrophoresis. amino acid analysis revealed high quantities of lys, his, ala, val, ile, glx, and pro residues. the circular dichroism revealed 45% beta-structure and practically lack of a-spiral domains. kinetic studies showe ... | 2010 | 20469643 |
| thermostable l-arabinose isomerase from bacillus stearothermophilus iam 11001 for d-tagatose production: gene cloning, purification and characterisation. | d-tagatose, as one of the rare sugars, has been found to be a natural and safe low-calorie sweetener in food products and is classified as a gras substance. l-arabinose isomerase (l-ai, ec 5.3.1.4), catalysing the isomerisations of l-arabinose and d-galactose to l-ribulose and d-tagatose respectively, is considered to be the most promising enzyme for the production of d-tagatose. | 2010 | 20474051 |
| cloning and characterization of a xylanase, krict px1 from the strain paenibacillus sp. hpl-001. | the krict px1 gene (gb: fj380951) consisting of 996bp encoding a protein of 332 amino acids (38.1kda) from the recently isolated paenibacillus sp. strain hpl-001 (kctc11365bp) has been cloned and expressed in escherichia coli. the xylanase krict px1 showed high activity on birchwood xylan, and was active over a ph range of 5.0 to 11.0, with two optima at ph 5.5 and 9.5 at 50 degrees c with k(m) value of 5.35 and 3.23, respectively. the xylanase activity was not affected by most salts, such as na ... | 2010 | 20493247 |