Publications

a few amino acid substitutions are responsible for the higher thermostability of a novel nad(+)-dependent bacillar alcohol dehydrogenase.	the gene adh-ht encoding a thermostable and thermophilic nad(+)-dependent alcohol dehydrogenase (adh) from the novel and more thermophilic bacillus stearothermophilus lld-r strain was cloned and its nucleotide sequence determined. the deduced protein sequence shows remarkable amino acid substitutions when compared to the sequence of the protein isolated from strain nca1503 and significant similarity with the highly thermostable adh from the thermoacidophilic archaebacterium sulfolobus solfataric ...	1994	8020473
sequence and analysis of the citrulline biosynthetic operon argc-f from bacillus subtilis.	the citrulline biosynthetic operon argc-f located at 100 degrees on the bacillus subtilis chromosome contains seven open reading frames which encode all the enzymes required for the biosynthesis of citrulline. the operon is transcribed as a single transcription unit. the second cistron of the operon is homologous to argj (ornithine acetyltransferase) from bacillus stearothermophilus and neisseria gonorrhoeae, suggesting that the acetylation of glutamate and the deacetylation of acetylornithine a ...	1994	8025667
cloning and dna sequence of the gene coding for bacillus stearothermophilus t-6 xylanase.	bacillus stearothermophilus t-6 produces an extracellular thermostable xylanase. affinity-purified polyclonal serum raised against the enzyme was used to screen a genomic library of b. stearothermophilus t-6 constructed in lambda-embl3. two positive phages were isolated, both containing similar 13-kb inserts, and their lysates exhibited xylanase activity. a 3,696-bp sali-bamhi fragment containing the xylanase gene was subcloned in escherichia coli and subsequently sequenced. the open reading fra ...	1994	8031084
proton and nitrogen nmr sequence-specific assignments and secondary structure determination of the bacillus subtilis spo1-encoded transcription factor 1.	sequence-specific 1h and 15n nmr1 assignments are reported for the transcription factor 1 (tf1), a 22-kda type ii dna-binding protein (dbpii) that consists of two 99-residue monomers. an assignment strategy is employed that uses six complementary selectively deuterium-labeled tf1 variants and an uniformly 15n-labeled tf1 variant. two-dimensional and three-dimensional homonuclear and heteronuclear nmr correlated spectra are analyzed and yield nearly complete assignments for the 1h and 15n resonan ...	1994	8038176
sequence analysis of the sbsa gene encoding the 130-kda surface-layer protein of bacillus stearothermophilus strain pv72.	bacillus stearothermophilus (bs) contains a surface-layer (s-layer) protein (sbsa), which forms a hexagonal array on the cell wall. in order to understand the structural/functional relationship of sbsa from bs pv72, the entire nucleotide (nt) sequence of the sbsa gene was determined from three overlapping fragments. the 3'-end was cloned and expressed in escherichia coli, whereas the 5'-region was amplified from the genome of bs pv72 by the polymerase chain reaction using two overlapping fragmen ...	1994	8045409
gene structure and amino acid sequences of alcohol dehydrogenases of bacillus stearothermophilus.	partial amino acid sequences of the two alcohol dehydrogenases of bacillus stearothermophilus and the oligonucleotide sequence of a cloned fragment containing the gene for adh 2334 were determined and compared with the known, derived adh 1503 amino acid sequence. the two proteins are identical at 244 of 349 positions. adh 2334 is encoded in a transcription unit containing an aldehyde dehydrogenase.	1994	8049268
effectiveness of two types of sterilizers on the contents of sharps containers.	the purpose of this study was to evaluate the killing effect that a gravity steam autoclave (gsa), a high-vacuum steam sterilizer (hva) or an unsaturated chemical vapor sterilizer (ucv) had on endospores present on strips or applied to dental needles within three sizes of sharps containers. commercial spore strips containing 1.7 x 10(5) bacillus stearothermophilus endospores were used, while the needles were soiled with an equal number of spores or with spores mixed with blood. needles were test ...	1994	8054195
stability and proton-permeability of liposomes composed of archaeal tetraether lipids.	liposomes composed of tetraether lipids originating from the thermoacidophilic archaeon sulfolobus acidocaldarius were analyzed for their stability and proton permeability from 20 degrees c up to 80 degrees c. at room temperature, these liposomes are considerably more stable and have a much lower proton permeability than liposomes composed of diester lipids originating from the mesophilic bacterium escherichia coli or the thermophilic bacterium bacillus stearothermophilus. with increasing temper ...	1994	8054346
purification procedure for bacterial translational initiation factors if2 and if3.	in bacteria the initiation of protein synthesis is a complex phenomenon in which specific proteins, termed initiation factors (ifs) if1, if2, and if3, are involved. notwithstanding the progress made in understanding their functions, the precise molecular mechanisms of action of these factors remain somewhat obscure. one reason for this lack of knowledge is the difficulty involved in purifying sufficient quantities of these proteins. we have developed a new procedure for purification of ifs from ...	1994	8054843
zinc chelation and structural stability of adenylate kinase from bacillus subtilis.	adenylate kinase from bacillus subtilis, like the enzyme from bacillus stearothermophilus, contains a structural zinc atom. cys153 in the enzyme from b. stearothermophilus, which is involved in the zinc coordination, is replaced in the adenylate kinase from b. subtilis by an aspartic acid residue. therefore, we were interested in establishing whether this difference has an impact on the structure, the metal chelation, and the overall stability of these proteins. we also were interested in determ ...	1994	8061005
parameters governing steam sterilization of deadlegs.	use of saturated steam for sterilization-in-place (sip) is limited by factors effecting displacement of air from deadlegs. effects of tube diameter, length, orientation and position within a deadleg were quantitatively studied by examining temperature profiles and rates of kill of bacillus stearothermophilus spores. tube diameter had the greatest effect on sterilization. for small diameter tubes, 0.4 cm inside diameter (id), air displacement was minimal and due mainly to diffusion. 8.8 cm long t ...	1994	8069515
identification and characterization of clustered genes for thermostable xylan-degrading enzymes, beta-xylosidase and xylanase, of bacillus stearothermophilus 21.	bacillus stearothermophilus 21 is a gram-positive, facultative thermophilic aerobe that can utilize xylan as a sole source of carbon. we isolated this strain from soil, purified its extracellular xylanase and beta-xylosidase, and analyzed the two-step degradation of xylan by these enzymes (t. nanmori, t. watanabe, r. shinke, a. kohno, and y. kawamura, j. bacteriol. 172:6669-6672, 1990). an escherichia coli transformant carrying a 4.2-kbp chromosomal segment of this bacterium as a recombinant pla ...	1994	8074507
complete amino acid sequence of ribosomal protein s14 from bacillus stearothermophilus and homology studies to other ribosomal proteins.	the complete amino acid sequence of protein s14 from the small subunit of bacillus stearothermophilus was determined by n-terminal sequence analysis and by sequencing of overlapping peptides obtained from enzymatic digestions. protein s14 consists of 60 amino acid residues with a molecular mass of 7148 da. it has a high content of basic amino acids and a predicted isoelectric point of 11.46. protein s14 contains two pairs of cysteines in the carboxyl-terminal region, presumably linked by two sul ...	1994	8076678
the expression of the bstvim gene from bacillus stearothermophilus v is restricted to vegetative cell growth.	the activity of bstvi dna methyltransferase was monitored during the sporulative cycle of bacillus stearothermophilus v. significant methylase activity was found only in bacteria growing vegetatively. this was confirmed by northern hybridization, which indicated that the bstvim gene was not transcribed in cells undergoing sporulation. supporting evidence came from experiments which demonstrated that the rna polymerase holoenzyme from these cells did not recognize the promoter elements upstream o ...	1994	8081498
detection of penicillin g in milk using a conductimetric method.	a highly sensitive method was developed that used conductance measurement for the detection of penicillin g in milk. the method is based on the inhibition by the antibiotic on the growth of bacillus stearothermophilus atcc 10149. the conductance change in pm indicator agar containing the bacterial spores was continuously monitored at 55 degrees c by a microbiological analyzer, and the detection time was delayed when penicillin g was present in the samples. the detection limit of the method for p ...	1994	8083409
purification and characterization of thermostable beta-n-acetylhexosaminidase of bacillus stearothermophilus ch-4 isolated from chitin-containing compost.	thermostable exochitinase was purified to homogeneity from the culture fluid of bacillus stearothermophilus ch-4, which was isolated from agricultural compost containing shrimp and crabs. the enzyme was a single polypeptide with a molecular mass of 74 kda, and the n-terminal amino acid sequence was wdkvgvtdli islnipeadavvvgmtlqlqalhly. the enzyme specifically hydrolyzed c-4 beta-anomeric bonding of n-acetylchitooligosaccharides, as well as their p-nitrophenyl (pnp) derivatives. the enzyme also h ...	1994	8085829
use of biological indicators designed for steam or ethylene oxide to monitor a liquid chemical sterilization process.	to determine the ability of a commercially available biological indicator (bi), used to monitor steam and ethylene oxide sterilization, to biologically monitor a liquid sterilization process consisting of a sterile processor, a proprietary peracetic acid sterilant, and a sterile rinse system.	1993	8093121
molecular cloning, sequencing, and transcriptional analysis of the groesl operon from bacillus stearothermophilus.	using a gene probe of the bacillus subtilis groel gene, a 7.3-kb hindiii fragment of chromosomal dna of bacillus stearothermophilus was cloned. sequencing of 2,309 bp led to the detection of two open reading frames in the order groes groel. primer extension studies revealed one potential transcription start site preceding the groesl operon, which was activated upon temperature upshift. northern blot (rna) analysis resolved two mrna species with lengths of 2.2 and 1.5 kb; rna slot-blot experiment ...	1993	8096841
structural dependence of post-translational modification and reductive acetylation of the lipoyl domain of the pyruvate dehydrogenase multienzyme complex.	the lipoyl domain of the dihydrolipoyl acetyltransferase (e2) component of the pyruvate dehydrogenase multienzyme complex is recognized specifically by the lipoylating enzyme(s) in the cell and by the pyruvate dehydrogenase (e1) component in the parent complex. highly conserved aspartic acid and alanine residues flank the lipoyl-lysine residue, on the n and c-terminal sides, respectively, in the sharp beta-turn in which the lipoyl-lysine residue is prominently displayed. a sub-gene encoding the ...	1994	8107106
the dsg gene of myxococcus xanthus encodes a protein similar to translation initiation factor if3.	the dsg mutants of myxococcus xanthus are defective in fruiting body development and sporulation, yet they grow normally. the deduced amino acid sequence of the dsg gene product is 50 and 51% identical to the amino acid sequence of translation initiation factor if3 of both escherichia coli and bacillus stearothermophilus, respectively. however, the dsg protein has a carboxy-terminal extension of 66 amino acids, which are absent from its e. coli and b. stearothermophilus homologs. the shine-dalga ...	1994	8113184
the myxococcus xanthus dsg gene product performs functions of translation initiation factor if3 in vivo.	the amino acid sequence of the dsg protein is 50% identical to that of translation initiation factor if3 of escherichia coli, the product of its infc gene. anti-e. coli if3 antibodies cross-react with the dsg protein. tn5 insertion mutations in dsg are lethal. when ample nutrients are available, however, certain dsg point mutant strains grow at the same rate as wild-type cells. under the starvation conditions that induce fruiting body development, these dsg mutants begin to aggregate but fail to ...	1994	8113185
source of catalysis in the lactate dehydrogenase system. ground-state interactions in the enzyme-substrate complex.	the raman spectra of both the nad-pyruvate and the pyridine aldehyde adenine dinucleotide (paad)-pyruvate bound to pig heart, pig muscle, and bacillus stearothermophilus lactate dehydrogenases were measured and are nearly the same, which is consistent with the conserved shell of residues surrounding the active-site cavity in these enzymes. the symmetrical stretching mode of the pyruvate carboxylate group, found at 1398 cm-1, is shifted only slightly when complexed to these enzymes, which shows t ...	1994	8117687
molecular recognition in proteins. simulation analysis of substrate binding by a tyrosyl-trna synthetase mutant.	alchemical molecular dynamics simulations are performed to determine the difference in the free energy of binding of the tyrosine substrate between the wild type of tyrosyl-trna synthetase (tyrrs) from bacillus stearothermophilus and the mutant tyr169-->phe. the results are of general interest because the tyr169 hydroxyl group interacts with the ammonium group of the substrate in a manner corresponding to that found in other amino acid binding proteins (e.g. the asp receptor of the chemotactic b ...	1994	8120886
involvement of conserved lysine 68 of bacillus stearothermophilus leucine dehydrogenase in substrate binding.	lysine 68 of bacillus stearothermophilus leucine dehydrogenase is highly conserved in the corresponding regions of nad(p)+-dependent amino acid dehydrogenase sequences. to elucidate its functional role, the lysyl residue of the recombinant enzyme has been replaced with alanine or arginine by site-directed mutagenesis. either mutation resulted in nearly complete loss of activity in the oxidative deamination, whereas only the mutation to alanine led to a marked increase in michaelis constants for ...	1994	8125938
[effect of microwaves on the vegetative and spore forms of bacillus stearothermophilus].	microwave irradiation provides a rapid and effective method for sterilization of stainless steel scalpel blades or cover glasses contaminated by b. stearothermophilus. a study by sem of vegetative forms showed that microwave irradiation induce a progressive series of alterations and finally the complete destruction of the microorganism. on the contrary there were no significant morphological variations of the spores after lethal irradiation by microwaves.	1993	8129884
the complete primary structure of ribosomal protein l1 from thermus thermophilus.	the primary structure of the 23s rrna binding ribosomal protein l1 from the 50s ribosomal subunit of thermus thermophilus ribosomes has been elucidated by direct protein sequencing of selected peptides prepared by enzymatic and chemical cleavage of the intact purified protein. the polypeptide chain contains 228 amino acids and has a calculated molecular mass of 24,694 d. a comparison with the primary structures of the corresponding proteins from escherichia coli and bacillus stearothermophilus r ...	1993	8136022
characterization of the two anion-recognition sites of glyceraldehyde-3-phosphate dehydrogenase from bacillus stearothermophilus by site-directed mutagenesis and chemical modification.	the active site of the glycolytic glyceraldehyde-3-phosphate dehydrogenase (gapdh) contains two anion recognition sites which have been attributed to the phosphate binding of the substrates, namely, glyceraldehyde 3-phosphate (ps site) and inorganic phosphate (pi site) [moras et al. (1975) j. biol. chem. 250, 9137-9162]. in order to probe the role of both sites during the catalytic event, arg 195 from the pi site and arg 231 from the ps site of the bacillus stearothermophilus enzyme have been ch ...	1994	8136361
kinetic characteristics of phosphofructokinase from bacillus stearothermophilus: mgatp nonallosterically inhibits the enzyme.	the kinetic mechanism of phosphofructokinase from bacillus sterothermophilus has been investigated using steady-state measurements. the double-reciprocal patterns observed for initial velocity, product inhibition, and mixed alternate substrate studies of the reverse reaction establish that the mechanism involves rapid-equilibrium random binding of substrates and the formation of an abortive complex composed of enzyme, mgadp, and fructose 6-phosphate (e-mgadp-fru-6p). initial velocity patterns fo ...	1994	8136379
protein stabilization by hydrophobic interactions at the surface.	the contribution of the solvent-exposed residue 63 to thermal stability of the thermolysin-like neutral protease of bacillus stearothermophilus was studied by analyzing the effect of twelve different amino acid substitutions at this position. the thermal stability of the enzyme was increased considerably by introducing arg, lys or bulky hydrophobic amino acids. in general, the effects of the mutations showed that hydrophobic contacts in this surface-located region of the protein are a major dete ...	1994	8143751
expression in escherichia coli of genes encoding the e1 alpha and e1 beta subunits of the pyruvate dehydrogenase complex of bacillus stearothermophilus and assembly of a functional e1 component (alpha 2 beta 2) in vitro.	the e1 alpha and e1 beta subunits of the pyruvate decarboxylase (e1) component of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus were produced from two genes overexpressed separately in escherichia coli. a functional e1 enzyme was generated from disrupted mixtures of cells containing the separately overexpressed e1 alpha and e1 beta genes. the purified e1 enzyme exhibited an apparent molecular mass of 150,000 da, consistent with an alpha 2 beta 2 structure. the km ...	1994	8144620
overcoming non-isomorphism by phase permutation and likelihood scoring: solution of the trprs crystal structure.	entropy maximization to maximum likelihood, constrained jointly by the best available experimental phases and by a sufficiently good envelope, can bring about substantial model-independent map improvement, even at medium (3.1 a) resolution [xiang, carter, bricogne & gilmore (1993). acta cryst. d49, 193-212]. in the crystal structure determination of the bacillus stearothermophilus tryptophanyl-trna synthetase (trprs), however, the following had to be dealt with simultaneously: (1) a serious lack ...	1994	8166950
quantitative analysis of crystal growth. tryptophanyl-trna synthetase crystal polymorphism and its relationship to catalysis.	we show that quantitative analysis of replicated, full-factorial crystal growth experiments and, by implication, similar studies of a wide variety of other phenomena, can be a powerful tool for analyzing macromolecular systems with complex, interacting dependencies on functionally significant factors. bacillus stearothermophilus tryptophanyl-trna synthetase crystallizes in three different crystal forms depending on the ligands present under otherwise identical conditions. comparison of crystallo ...	1994	8176729
allosteric activation in bacillus stearothermophilus lactate dehydrogenase investigated by an x-ray crystallographic analysis of a mutant designed to prevent tetramerization of the enzyme.	the crystal structure of a mutant bacillus stearothermophilus lactate dehydrogenase, into which an additional loop has been engineered in order to prevent tetramerization of the enzyme, has been solved and refined at 2.4 a. the minimal repeat unit in the crystal is a dimer and the tetramer cannot be generated by any of the crystallographic symmetry operations in p2(1). the loop protrudes out into the solvent, stabilized by a good hydrogen bonding arrangement, and clearly sterically hinders tetra ...	1994	8176749
structure-function relationships in the catalytic and starch binding domains of glucoamylase.	sixteen primary sequences from five sub-families of fungal, yeast and bacterial glucoamylases were related to structural information from the model of the catalytic domain of aspergillus awamori var. x100 glucoamylase obtained by protein crystallography. this domain is composed of thirteen alpha-helices, with five conserved regions defining the active site. interactions between methyl alpha-maltoside and active site residues were modelled, and the importance of these residues on the catalytic ac ...	1994	8177888
comparative cell targeting in vitro using the co2 laser.	in keeping with current interest in bactericidal effects of lasers to treat localized infection, we examined the effect of co2 laser light on three bacterial species. cell monolayers were placed atop filters minimizing the presence of extracellular water. samples were exposed to irradiance values between 0 and 520 w/cm2 for 0.1 s. cell survival was determined by colony counts. there was logarithmic killing of the bacteria with increasing irradiance; 90% kill was effected at 180 w/cm2 for escheri ...	1994	8183050
molecular cloning of a maltose transport gene from bacillus stearothermophilus and its expression in escherichia coli k-12.	genes responsible for maltose utilization from bacillus stearothermophilus atcc7953 were cloned in the plasmid vector pbr325 and functionally expressed in escherichia coli. the 4.2 kb bacillus dna insert in clone pam1750 suppressed the growth defects on maltose caused by mutations in e. coli maltose transport genes (male, malk or complete malb deletion) but not mutations in genes affecting intracellular maltose metabolism (mala region). transport studies in e. coli and b. stearothermophilus sugg ...	1994	8190087
discrimination between transfer-rnas by tyrosyl-trna synthetase.	we have constructed a model of the complex between tyrosyl-trna synthetase (tyrrs) from bacillus stearothermophilus and trna(tyr) by successive cycles of predictions, mutagenesis of tyrrs and molecular modeling. we confront this model with data obtained independently, compare it to the crystal structures of other complexes and review recent data on the discrimination between trnas by tyrrs. comparison of the crystal structures of tyrrs and glnrs, both of which are class i synthetases, and compar ...	1993	8199245
regulation of dihydrodipicolinate synthase and diaminopimelate decarboxylase activity in bacillus stearothermophilus.	the feedback inhibition of the enzymes dihydrodipicolinate (dhdps) and diaminopimelate decarboxylase (dapd) in the wild strain zu 183 of bacillus stearothermophilus and in its s-(2-aminoethyl)-cysteine resistant l-lysine overproducing strain aec 12 was studied. the optimum temperature and ph of both enzymes were also evaluated. no inhibition of dhdps by l-lysine, l-threonine, l-methionine and l-isoleucine was observed either in the wild strain or in the aec 12 mutant. dapd was completely inhibit ...	1994	8206742
immunoelectron microscopic localization of ribosomal proteins bs8, bs9, bs20, bl3 and bl21 on the surface of 30s and 50s subunits from bacillus stearothermophilus.	the locations of ribosomal proteins bs8, bs9 and bs20 on the 30s subunit of bacillus stearothermophilus ribosomes, and of bl3 and bl21 on the 50s subunit, were determined by immunoelectron microscopy. bl3 was found to lie half-way down the body of the 50s subunit on the interface side, below the l7/l12 stalk, in agreement with the placement of the corresponding protein in escherichia coli by neutron-scattering; bl21 was located at a similar position on the solvent side of the subunit, as predict ...	1993	8223574
cloning, purification and characterization of the bseci dna methyltransferase from bacillus stearothermophilus.	the gene (bsecim) encoding the bseci dna methyltransferase (mtase; m.bseci) from a bacillus stearothermophilus species was cloned and expressed in escherichia coli using plasmid vector pbr322. selection of transformants carrying bsecim was based on the resistance of the modified plasmid to cleavage by bseci. the mtase was purified to homogeneity and further characterized. its size as determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis and size exclusion chromatography was 68 ...	1993	8224900
evaluation of milk antibiotic residue screening tests in cattle with naturally occurring clinical mastitis.	milk from 172 commercial cows with mild to moderate clinical mastitis was tested with five antibiotic residue detection assay systems. one hundred cows were treated with one of two intramammary beta-lactam antibiotics, and the remaining 72 cows were treated with intramuscular oxytocin. milk samples were collected pretreatment, twice after therapy, and again 21 d following the initiation of treatment. presumptive false-positive assay results were tabulated from all pretreatment and 21-d milk samp ...	1993	8227629
mapping the stability determinants of bacterial tyrosyl transfer rna synthetases by an experimental evolutionary approach.	the tyrosyl-trna synthetases from bacillus stearothermophilus (bst-tyrts) and escherichia coli (eco-tyrts) are 56% identical in amino acid sequence. to map and characterize the set of interactions that makes bst-tyrts more stable than eco-tyrts, a family of nine hybrid proteins was constructed between the two enzymes. the n-terminal part of each hybrid came from eco-tyrts and the c-terminal part from bst-tyrts. the stability and activity of these hybrids were estimated by experiments of thermal ...	1993	8230200
sequence of the triosephosphate isomerase-encoding gene isolated from the thermophile bacillus stearothermophilus.	by analysis of genomic clones, we have determined the complete nucleotide sequence of the gene encoding triosephosphate isomerase (tim; ec 5.3.1.1) in the thermophilic bacterium, bacillus stearothermophilus. the gene encodes a 253-amino-acid tim which is 39% identical to that of the mesophile, escherichia coli.	1993	8244026
threonine 246 at the active site of the l-lactate dehydrogenase of bacillus stearothermophilus is important for catalysis but not for substrate binding.	threonine 246 is an active site residue that is conserved in all known l-lactate dehydrogenase (ldh; ec 1.1.1.27) sequences. in order to investigate the role of thr246 in bacillus stearothermophilus ldh, this residue was altered by site-directed mutagenesis to valine, alanine, leucine, and serine, respectively. the effects of these mutations, as observed in both steady-state and single-turnover kinetic measurements with different substrates, demonstrated the importance for catalysis of a hydroxy ...	1993	8251493
ribosomal protein s17: characterization of the three-dimensional structure by 1h and 15n nmr.	the structure of ribosomal protein s17 from bacillus stearothermophilus was investigated by two-dimensional homonuclear and heteronuclear magnetic resonance spectroscopy. the 1h and 15n chemical shift assignments are largely complete, and a preliminary structural characterization is presented. the protein consists of five beta-strands that form a single antiparallel beta-sheet with greek-key topology. the beta-strands are connected by several extended loops, and two of these contain residue type ...	1993	8251502
involvement of threonine 234 in catalysis of tyrosyl adenylate formation by tyrosyl-trna synthetase.	there is a mobile loop in the tyrosyl-trna synthetase that contains the kmsks signature sequence of class i aminoacyl-trna synthetases. as it has not been possible to determine the role of the mobile loop in catalysis from x-ray crystallographic studies, we are investigating its importance by a series of site-directed mutagenic and kinetic studies. here we examine the role of threonine 234 (t234) in the catalysis of tyrosyl adenylate formation by tyrosyl-trna synthetase from bacillus stearotherm ...	1993	8257697
mutation of lysine 233 to alanine introduces positive cooperativity into tyrosyl-trna synthetase.	tyrosyl-trna synthetase from bacillus stearothermophilus is a dimeric enzyme which displays half-of-sites reactivity with respect to the binding of both tyrosine and atp. the binding of both substrates follows michaelis-menten kinetics. mutation of lysine 233 to alanine (k233a) decreases the affinity of the active subunit for atp at both saturating and subsaturating tyrosine concentrations (from the hill plot, kcat = 0.56 s-1, nh = 1.54, kd = 372 mm at 50 microm tyrosine). in addition, this muta ...	1993	8257698
ribosomal protein l6: structural evidence of gene duplication from a primitive rna binding protein.	in all cells, protein synthesis is coordinated by the ribosome, a large ribonucleoprotein particle that is composed of > 50 distinct protein molecules and several large rna molecules. here we present the crystal structure of ribosomal protein l6 from the thermophilic bacterium bacillus stearothermophilus solved at 2.6 a resolution. l6 contains two domains with almost identical folds, implying that it was created by an ancient gene duplication event. the surface of the molecule displays several l ...	1993	8262035
evidence for lysine 80 as general base catalyst of leucine dehydrogenase.	to elucidate the functional role of the lysyl residue highly conserved in nad(p)(+)-dependent amino acid dehydrogenases, lys-80 of leucine dehydrogenase from bacillus stearothermophilus has been mutated into ala, arg, or gln. all of the mutant enzymes had markedly reduced activities in the oxidative deamination, whereas the michaelis constants for substrate and coenzyme did not change significantly upon the mutation, except for a 10-30-fold increase in km values for alpha-keto-iso-caproate in th ...	1993	8262941
temperature-induced inversion of allosteric phenomena.	two instances, involving the enzymes carbamoyl-phosphate synthetase from escherichia coli and phosphofructokinase from bacillus stearothermophilus, respectively, are described in which increasing temperature alone causes the actions of an allosteric ligand to change from inhibition to activation. in neither case are these effects due to a change in the activation energy of the enzyme catalyzed reaction induced by the allosteric ligand. rather, they are due to temperature-dependent changes in the ...	1994	8276837
the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the pyruvate dehydrogenase complex of bacillus stearothermophilus: preparation and characterization of its binding to the dihydrolipoyl dehydrogenase component.	the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase polypeptide chain of the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus was released by limited proteolysis from a di-domain (lipoyl domain plus binding domain) encoded by a subgene over-expressed in escherichia coli. the domain was characterized by n-terminal sequence analysis, electrospray m.s. and c.d. spectroscopy. it was found to be identical in all respects to a chemically synthesized pe ...	1994	8280091
gene cloning, sequence analysis, purification, and characterization of a thermostable aminoacylase from bacillus stearothermophilus.	a genomic dna fragment encoding aminoacylase activity of the eubacterium bacillus stearothermophilus was cloned into escherichia coli. transformants expressing aminoacylase activity were selected by their ability to complement e. coli mutants defective in acetylornithine deacetylase activity, the enzyme that converts n-acetylornithine to ornithine in the arginine biosynthetic pathway. the 2.3-kb cloned fragment has been entirely sequenced. analysis of the sequence revealed two open reading frame ...	1993	8285691
studies on thermophile products. vi. activation of mouse peritoneal macrophages by bis(2-hydroxyethyl) trisulfide.	the biological effects of a cytotoxic substance (bs-1), isolated from bacillus stearothermophilus uk563 and identified as bis(2-hydroxyethyl) trisulfide, on elicited mouse peritoneal macrophages induced by a casein injection, were investigated in vitro. balb/c mouse macrophages treated or pretreated with bs-1 (1-10 micrograms/ml) showed cytotoxicity against syngeneic dba/2 mouse p815 mastocytoma. bs-1 also showed weak cytotoxicity directly against p815 in the absence of macrophages. bs-1 signifi ...	1993	8287048
zinc, a structural component of adenylate kinases from gram-positive bacteria.	the recent finding that bacillus stearothermophilus adenylate kinase contains a zinc atom coordinated to four cysteines prompted us to investigate the metal-binding properties of the enzyme from various bacteria. we conclude that zinc was present only in adenylate kinase from gram-positive species and that this property is correlated with the presence of three or four cys residues in the sequence cys-x2-cys-x16-cys-x2-cys/asp, in which x stands for different amino acid residues.	1994	8288548
isolation of two physiologically induced variant strains of bacillus stearothermophilus nrs 2004/3a and characterization of their s-layer lattices.	during growth of bacillus stearothermophilus nrs 2004/3a in continuous culture on complex medium, the chemical properties of the s-layer glycoprotein and the characteristic oblique lattice were maintained only if glucose was used as the sole carbon source. with increased aeration, amino acids were also metabolized, accompanied by liberation of ammonium and by changes in the s-layer protein. depending on the stage of fermentation at which oxygen limitation was relieved, two different variants, on ...	1994	8300538
crystal structure of prokaryotic ribosomal protein l9: a bi-lobed rna-binding protein.	the crystal structure of protein l9 from the bacillus stearothermophilus ribosome has been determined at 2.8 a resolution using x-ray diffraction methods. this primary rna-binding protein has a highly elongated and unusual structure consisting of two separated domains joined by a long exposed alpha-helix. conserved, positively charged and aromatic amino acids on the surfaces of both domains probably represent the sites of specific interactions with 23s rrna. comparisons with other prokaryotic l9 ...	1994	8306963
zinc ions bound to chimeric his4/lactate dehydrogenase facilitate decarboxylation of oxaloacetate.	a chemically synthesized dna linker coding for a peptide fragment that contains four histidines was fused in-frame to the 5'-end of the bacillus stearothermophilus lactate dehydrogenase gene. the gene product, his4/lactate dehydrogenase, could be purified to homogeneity using either immobilized metal (zn2+)-affinity chromatography or affinity chromatography on oxamate agarose. the stability against heat and urea for the modified enzymes was decreased as compared to the native lactate dehydrogena ...	1993	8309939
comparison of the inhibition by phospho(enol)pyruvate and phosphoglycolate of phosphofructokinase from b. stearothermophilus.	a comparison between the inhibition by phospho(enol)pyruvate (pep) versus the inhibition by phosphoglycolate (pg) of phosphofructokinase (pfk) from bacillus stearothermophilus is presented. both inhibitors act by decreasing the apparent affinity displayed by the enzyme for its substrate fructose 6-phosphate (fru-6-p) while having little effect on vmax. however, the two ligands differ in both their affinity for the enzyme and their effectiveness at antagonizing the subsequent binding of fru-6-p. ...	1994	8311457
two putative insertion sequences flank a truncated glycogen branching enzyme gene in the thermophile bacillus stearothermophilus cu21.	we have isolated a region from the bacillus stearothermophilus cu21 chromosome hybridizing strongly to a fragment of the b. caldolyticus glycogen operon. sequence analysis of this region revealed the presence of a truncated glgb gene encoding the n-terminus of branching enzyme. a region highly similar to an internal fragment of b. caldolyticus glgc encoding adp-glucose pyrophosphorylase was located approximately 1kb downstream from the incomplete glgb gene. the two truncated genes appeared to fl ...	1993	8312600
purification and characterization of a thermostable xylanase from bacillus stearothermophilus t-6.	bacillus stearothermophilus t-6 produces an extracellular xylanase that was shown to optimally bleach pulp at ph 9 and 65 degrees c. the enzyme was purified and concentrated in a single adsorption step onto a cation exchanger and is made of a single polypeptide with an apparent m(r) of 43,000 (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). xylanase t-6 is an endoxylanase that completely degrades xylan to xylose and xylobiose. the pis of the purified protein were 9 and ...	1993	8328796
the relationships between transketolase, yeast pyruvate decarboxylase and pyruvate dehydrogenase of the pyruvate dehydrogenase complex.	the amino acid sequences of four thiamine pyrophosphate-requiring enzymes were aligned with the published amino acid sequence of the transketolase of hansenula polymorpha. sequences of the combined alpha and beta subunits of the e1 enzyme of the pyruvate dehydrogenase complexes of homo sapiens and bacillus stearothermophilus aligned well with the transketolase while the e1 of the pyruvate dehydrogenase complex of escherichia coli aligned easily provided a non-aligning segment of 77 amino acids w ...	1993	8344439
[isolation and properties of bstbsi restriction endonuclease from the thermophilic soil bacteria bacillus stearothermophilus bs].	a new restriction endonuclease bstbsi was isolated and purified from the thermophilic soil bacterium bacillus stearothermophilus bs by the blue sepharose and hydroxyapatite chromatographies. the enzyme is an isoschizomer of snai from sphaerotilus natans c. it recognizes the hexanucleotide gtatac and cleaves dna in the center of the sequence. the maximal catalytic activity of the endonuclease is registered in 50 mm tris-hcl (ph 9.0) buffer with the high ionic strength (100 mm nacl) in the presenc ...	1993	8350878
evaluation of a rapid readout biological indicator for flash sterilization with three biological indicators and three chemical indicators.	flash sterilization is most commonly used for emergency sterilization of unwrapped items in a gravity displacement sterilizer for three minutes. sterilization quality assurance is monitored by biological indicators that require a 24-hour incubation prior to reading. in this study, we compared a new biological indicator that provides results within 60 minutes with three conventional, 24-hour biological indicators for monitoring flash sterilization and three chemical indicators.	1993	8354870
cloning, sequencing, and expression in escherichia coli of the gene coding for phosphofructokinase in lactobacillus bulgaricus.	a fragment of 1,185 bp containing the gene coding for phosphofructokinase (atp:d-fructose-6-phosphate-1-phosphotransferase; ec 2.7.1.11) in lactobacillus bulgaricus has been cloned, sequenced, and expressed in escherichia coli. the amino acid sequence of this enzyme was homologous to those of the atp-dependent phosphofructokinases from e. coli, thermus thermophilus, spiroplasma citri, and bacillus stearothermophilus, suggesting that these enzymes have closely related structures despite their dif ...	1993	8366023
the primary structure of a fungal chitin deacetylase reveals the function for two bacterial gene products.	chitin deacetylase (ec 3.5.1.41) hydrolyzes the n-acetamido groups of n-acetyl-d-glucosamine residues in chitin. a cdna to the mucor rouxii mrna encoding chitin deacetylase was isolated, characterized, and sequenced. protein sequence comparisons revealed significant similarities of the fungal chitin deacetylase to rhizobial nodb proteins and to an uncharacterized protein encoded by a bacillus stearothermophilus open reading frame. these data suggest the functional homology of these evolutionaril ...	1993	8367456
time-resolved fluorescence of the single tryptophan of bacillus stearothermophilus phosphofructokinase.	the fluorescence of the single tryptophan in bacillus stearothermophilus phosphofructokinase was characterized by steady-state and time-resolved techniques. the enzyme is a tetramer of identical subunits, which undergo a concerted allosteric transition. time-resolved emission spectral data were fitted to discrete and distributed lifetime models. the fluorescence decay is a double exponential with lifetimes of 1.6 and 4.4 ns and relative amplitudes of 40 and 60%. the emission spectra of both comp ...	1993	8369432
ribosomal proteins, tl4 and tl5, from thermus thermophilus form hybrid complexes with 5 s ribosomal rna from different microorganisms.	hybrid complexes of the ribosomal proteins, tl4 and tl5, from thermus thermophilus with 5 s ribosomal rna from escherichia coli and bacillus stearothermophilus have been prepared. there was no competition between the two proteins for the binding sites. stoichiometry of 5 s rna binding for both proteins was 1:1 (protein/rna). the tl4 protein competed with the e. coli ribosomal l5 protein, and the tl5 protein competed with the e. coli ribosomal proteins, l18 and l25, for binding with 5 s rna.	1993	8370456
characterization of the bstvirm genes encoding the bacillus stearothermophilus v restriction-modification system.	the nucleotide (nt) sequence of a 2.7-kb hindiii-ecori dna fragment encoding the bstvir and bstvim genes has been determined. the sequence predicts a restriction endonuclease of 224 amino acids (aa), m(r) 25,104, and a methyl-transferase of 561 aa, m(r0 65,702. both genes are aligned in the same orientation and are separated by a 102-nt intergenic region. no homology was found between r.bstvi and m.bstvi when their deduced aa sequences were compared. significant similarity at the aa level was fo ...	1993	8370531
the daca gene of bacillus stearothermophilus coding for d-alanine carboxypeptidase: cloning, structure and expression in escherichia coli and pichia pastoris.	the bacterial d-alanine carboxypeptidases (cpases) remove c-terminal d-alanyl residues from sugar-peptide cell wall precursors. the cpases have many characteristics in common with the high-m(r) penicillin-binding proteins (pbps) whose inhibition by beta-lactam antibiotics is lethal. the cpases are attractive as model pbps, because of their relatively lower m(r) and higher activity in vitro. we have cloned and sequenced the bacillus stearothermophilus gene (daca) coding for a membrane-bound cpase ...	1993	8370539
identification of the l-tartrate dehydratase genes (ttda and ttdb) of escherichia coli and evolutionary relationship with the class i fumarase genes.	the genes encoding an oxygen-labile stereospecific l-tartrate dehydratase (l-ttd, ec 4.2.1.32) have been identified as the orfz1 and orfz2 genes located upstream of the rpsu-dnag-rpod operon at 67 min in the escherichia coli linkage map. they were previously cloned and sequenced by m. nesin and others (gene 51, 149-161, 1987) and have now been independently cloned, partially resequenced, and designated as an operon (ttdab) containing two translationally coupled genes. the enzyme behaves as a tet ...	1993	8371115
dicranin, an antimicrobial and 15-lipoxygenase inhibitor from the moss dicranum scoparium.	extracts of nine mosses, collected in switzerland, were screened for antimicrobial, antioxidative, and 15-lipoxygenase (15-lpo) inhibitory activities. the ch2cl2 extract of dicranum scoparium was found to possess pronounced antimicrobial activity against bacillus cereus, bacillus stearothermophilus, bacillus subtilis, staphylococcus aureus, and escherichia coli. in addition, inhibition of soybean 15-lpo occurred at very low concentration. phytochemical investigation of this extract afforded z,z, ...	1993	8377015
on two transposable elements from bacillus stearothermophilus.	two transposable elements, is5376 and is5377, were identified in the thermophile bacillus stearothermophilus cu21 based upon the following criteria: (1) both were found to appear on different plasmids introduced into the same host cu21; (2) signals of homology were found between the genomic dna of cu21 and each of them; (3) different numbers of southern hybridization bands were found for the genomic dna of different strains of b. stearothermophilus; and (4) characteristic inverted repeats at bot ...	1993	8382825
energy-transducing properties of primary proton pumps reconstituted into archaeal bipolar lipid vesicles.	archaeal lipids differ considerably from eubacterial and eukaryotic lipids in their structure and physical properties. from the membranes of the extreme thermophilic archaea sulfolobus acidocaldarius a tetraether lipid fraction was isolated, which can form closed and stable monolayer liposomes in aqueous media. the function of three different primary proton pumps originating from archaeal, bacterial and eukaryotic lipid sources have been studied after reconstitution in these liposomes: bacterior ...	1993	8391438
rat muscle 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. study of the kinase domain by site-directed mutagenesis.	sequence alignment and modeling of the 2-kinase domain of the liver bifunctional enzyme, 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase, on 6-phosphofructo-1-kinase from bacillus stearothermophilus and escherichia coli (bazan, j. f., fletterick, r. j., and pilkis, s. j. (1989) proc. natl. acad. sci. u.s.a. 86, 9642-9646) suggested that cys-160 of the 2-kinase would correspond to asp-127 of the 1-kinase, which acts as a general base catalyst. we have studied the validity of this alignment ...	1993	8392072
[n-acetylglutamate-5-phosphotransferase of the thermophilic bacterium bacillus stearothermophilus: nucleotide sequence of the gene and enzyme characterization].	a nucleotide sequence of the argb gene of strain bacillus stearothermophilus ncib 8224 was determined. the argb gene codes for n-acetylglutamate-5-phosphotransferase of 258 amino acids with a molecular weight of 26918 d. this value is in good agreement with the sds-pag electrophoresis gata for identification of the heat stable b. stearothermophilus argb product synthesized in mesophilic escherichia coli host cells. the substrates mgatp and n-acetyl-l-glutamate efficiently protect the enzyme agai ...	1993	8394836
determinants of coenzyme specificity in glyceraldehyde-3-phosphate dehydrogenase: role of the acidic residue in the fingerprint region of the nucleotide binding fold.	on the basis of the three-dimensional structure of the glycolytic nad-dependent glyceraldehyde-3-phosphate dehydrogenase (gapdh) and of sequence comparison with the photosynthetic nad(p)-dependent gapdh of the chloroplast, a series of mutants of gapdh from bacillus stearothermophilus have been constructed. the results deduced from kinetic and binding studies suggest that the absence of activity of the wild-type gapdh with nadp as a cofactor is the consequence of at least three factors: (1) steri ...	1993	8399144
the identification of a lysine residue reactive to pyridoxal-5-phosphate in the glycerol dehydrogenase from the thermophile bacillus stearothermophilus.	the glycerol dehydrogenase (gdh) from bacillus stearothermophilus is inactivated by incubation with pyridoxal-5-phosphate (palp). the complex formed between the two can be trapped by reduction with sodium borohydride to yield a protein with an absorbance band at 325 nm and a fluorescence emission band at 430 nm, typical of trapped pyridoxal-5-phosphate moieties. total loss of catalytic activity of the enzyme is associated with the modification of approximately one equivalent of the reagent; the ...	1993	8399385
isolation and amino acid sequence of the 30s ribosomal protein s19 from mycobacterium bovis bcg.	the 30s ribosomal proteins from mycobacterium bovis bcg were separated by reverse phase-high performance liquid chromatography (rp-hplc). the isolated proteins were analyzed by sds-page, blotted on pvdf-membranes and subjected to sequence analyses using a gas-phase sequencer to correlate them to those of the well studied escherichia coli and bacillus stearothermophilus ribosomes. moreover, the internal amino acid sequence of one ribosomal protein, mbos19, which is homologous to e. coli ribosomal ...	1993	8405418
molecular cloning and sequencing of infc, the gene encoding translation initiation factor if3, from four enterobacterial species.	translation initiation factor if3 plays a crucial role in initiation of protein synthesis in bacteria. in order to elucidate the if3 structural elements required for these functions, the evolutionary conservation of if3 and its gene, infc, was investigated. homologous infc sequences from salmonella typhimurium, klebsiella pneumoniae, serratia marcescens and proteus vulgaris were amplified by the polymerase chain reaction and sequenced. analysis of these sequences, as well as that from bacillus s ...	1993	8405963
cloning and sequencing of a cellobiose phosphotransferase system operon from bacillus stearothermophilus xl-65-6 and functional expression in escherichia coli.	cellulolytic strains of bacillus stearothermophilus were isolated from nature and screened for the presence of activities associated with the degradation of plant cell walls. one isolate (strain xl-65-6) which exhibited strong activities with 4-methylumbelliferyl-beta-d-glucopyranoside (mug) and 4-methylumbelliferyl-beta-d-cellobiopyranoside (muc) was used to construct a gene library in escherichia coli. clones degrading these model substrates were found to encode the cellobiose-specific genes o ...	1993	8407820
site-directed mutagenesis of a hexapeptide segment involved in substrate recognition of phenylalanine dehydrogenase from thermoactinomyces intermedius.	phenylalanine dehydrogenase from thermoactinomyces intermedius and leucine dehydrogenase from bacillus stearothermophilus show a 59% sequence similarity in their substrate-binding domains, although their substrate specificities are different. we prepared a phenylalanine dehydrogenase mutant enzyme whose inherent hexapeptide segment (124f-v-h-a-a-129r) in the substrate-binding domain was replaced by the corresponding part of leucine dehydrogenase (m-d-i-i-y-q) in order to investigate the mechanis ...	1993	8407879
crystallization and preliminary x-ray diffraction studies of bacillus stearothermophilus farnesyl diphosphate synthase expressed in escherichia coli.	thermostable farnesyl diphosphate synthase (ec 2.5.1.10) from bacillus stearothermophilus, which was overexpressed in escherichia coli, has been crystallized by the vapor-diffusion procedure. tetragonal crystals were obtained using ammonium sulfate as a precipitant. the crystals diffracted x-rays to about 3 a resolution. the diffraction pattern indicated that the space group is i4(1)22 with unit-cell dimensions of a = b = 114 a and c = 247 a. it is thought that the asymmetric unit comprises two ...	1993	8411181
functional analysis of the lactococcus lactis usp45 secretion signal in the secretion of a homologous proteinase and a heterologous alpha-amylase.	the ups45 gene encodes the major extracellular protein from lactococcus lactis. the deduced sequence of the 27 residue leader peptide revealed the tripartite characteristics of a signal peptide. this leader peptide directed the efficient secretion of the homologous proteinase (prtp) in l. lactis, indicating that the putative signal peptide of prtp can be replaced by the 27 residue usp45 leader peptide. in addition, the 27 residue leader peptide could be used to secrete the bacillus stearothermop ...	1993	8413193
cloning and overexpression of the triosephosphate isomerase genes from psychrophilic and thermophilic bacteria. structural comparison of the predicted protein sequences.	we focused on the temperature adaptation of triosephosphate isomerase (tim; e.c. 5.3.1.1.) by comparing the structure of tims isolated from bacterial organisms living in either cold or hot environments. the tim gene from psychrophilic bacteria moraxella sp. ta137 was cloned and its nucleotide sequence determined. its deduced amino acid sequence revealed 34% identity with the thermophilic bacteria bacillus stearothermophilus tim. expression vectors were constructed and recombinant moraxella ta137 ...	1993	8421318
purification and characterisation of bst lvi restriction endonuclease, a thermostable isoschizomer of clai from bacillus stearothermophilus lv.	this work describes the purification and biochemical characterization of bstlvi restriction endonuclease, a thermostable isoschizomer of clai, from bacillus stearothermophilus lv. the enzyme was purified by successive deae-cellulose, affi-gel blue and heparin-sepharose cl-6b column chromatography. a molecular weight of 37,000 was determined for bst lvi by gel filtration. as expected from thermophilic proteins, the enzyme showed a high stability towards heat and also to other known protein-denatu ...	1993	8424953
stabilization of bacillus stearothermophilus neutral protease by introduction of prolines.	the thermostability of neutral proteases has been shown to depend on autolysis which presumably occurs in flexible regions of the protein. in an attempt to rigidify such a region in the neutral protease of bacillus stearothermophilus, residues in the solvent-exposed 63-69 loop were replaced by proline. the mutations caused large positive (ser-65-->pro, ala-69-->pro) or negative (thr-63-->pro, tyr-66-->pro) changes in thermostability, which were explained on the basis of molecular modelling of th ...	1993	8428638
molecular cloning and nucleotide sequence of the gene for pyruvate kinase of bacillus stearothermophilus and the production of the enzyme in escherichia coli. evidence that the genes for phosphofructokinase and pyruvate kinase constitute an operon.	pyruvate kinase from bacillus stearothermophilus is an allosteric enzyme activated by amp or ribose 5-phosphate but not by fructose 1,6-bisphosphate. the gene for the enzyme was cloned in escherichia coli and its entire nucleotide sequence was determined. the deduced amino acid sequence consisted of 587 residues and the molecular mass was calculated to be 62 317 da. the sequence was highly similar to other pyruvate kinases, indicating that they have the same evolutional origin. similarly to the ...	1993	8436141
effect of mutation of an amino acid residue near the catalytic site on the activity of bacillus stearothermophilus alpha-amylase.	site-directed mutagenesis of a thermostable alpha-amylase from bacillus stearothermophilus was performed to assess the role of amino acid residues near the catalytic site in catalysis. asn329 is presumed to be adjacent to the proposed catalytic residue asp331. its mutation to lys, which is found at the corresponding position in pullulanase, resulted in the loss of 99.7% of the activity, while the mutation to asp or val did not drastically reduce the activity. the mutation to val altered the temp ...	1993	8436143
modification of the amino acid specificity of tyrosyl-trna synthetase by protein engineering.	the amino acid specificity of bacillus stearothermophilus tyrosyl-trna synthetase was studied by site-directed mutagenesis of residues close to the active site. x-ray crystallographic studies of the enzyme have suggested that asp-176 is a major determinant of amino acid specificity, as its carboxylate is observed to make a hydrogen bond with the hydroxyl group of the substrate tyrosine. previous efforts to test the importance of asp-176 by site-directed mutagenesis led to inactive enzymes. we ha ...	1993	8440372
effects of steam sterilization on the contents of sharps containers.	one form of medical waste known to be capable of transmitting disease is the contaminated sharp. safe handling and disposal of sharps is an essential element of any infection control program. many areas allow the on-site treatment of sharps containers. however, little information currently exists as to the most effective sterilization procedures and container designs.	1993	8442519
dissecting the contributions of a specific side-chain interaction to folding and catalysis of bacillus stearothermophilus lactate dehydrogenase.	x-ray crystallography predicts hydrogen-bonding interactions between the side chains of thr198 and two other amino acid residues, glu194 (adjacent to the catalytic his195) and ser318 (on the alpha-h helix which rearranges on substrate binding). in order to investigate the contribution of this conserved amino acid residue, thr198, two mutants of bacillus stearothermophilus lactate dehydrogenase were created (val198 and ile198). the steady-state kinetic parameters for both mutant enzymes were very ...	1993	8444183
cross-linked amino acids in the protein pairs l3-l19 and l23-l29 of bacillus stearothermophilus ribosomes after treatment with diepoxybutane.	treatment of native 50 s ribosomal subunits of bacillus stearothermophilus with the homobifunctional cross-linking reagent diepoxybutane generated two cross-linked protein pairs, l3-l19 and l23-l29, which were isolated and identified. the analysis of the cross-linking sites at the amino acid level in both protein pairs is presented. using a combination of sequence analysis and mass spectrometry it could be demonstrated that his-28 in protein l3 and the n-terminal amino acids met-1, his-2, and hi ...	1993	8444837
three-dimensional structure of the lipoyl domain from bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex.	the structure of the lipoyl domain from the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been determined by means of nuclear magnetic resonance spectroscopy. a total of 452 nuclear overhauser effect distance constraints and 76 dihedral angle restraints were employed as the input for the structure calculations, which were performed using a hybrid distance geometry-simulated annealing strategy and the programs disgeo and x-plor. the overall structure of the lipoyl ...	1993	8445635
molecular basis of allosteric activation of bacterial l-lactate dehydrogenase.	the three-dimensional structure of allosteric l-lactate dehydrogenase from bifidobacterium longum, the first example of a t-state structure of l-lactate dehydrogenase, has been determined to 2.0 a. a comparative study of this structure with the previously reported r-state structure from bacillus stearothermophilus has revealed the allosteric activation mechanism of the bacterial l-lactate dehydrogenase. the fructose 1,6-bisphosphate-induced conformational change at the effector site and the subs ...	1993	8450537
the high-resolution structure of the peripheral subunit-binding domain of dihydrolipoamide acetyltransferase from the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus.	the three-dimensional structure of a 43-residue active, synthetic peptide encompassing the peripheral subunit-binding domain of dihydrolipoamide acetyltransferase from the pyruvate dehydrogenase multienzyme complex of bacillus stearothermophilus has been determined by means of a multi-cooling dynamical simulated annealing protocol using restraints derived from 1h nuclear magnetic resonance spectroscopy. a total of 442 experimentally derived restraints including 13 dihedral angle (phi, chi 1) res ...	1993	8450544
the structure of a thermally stable 3-phosphoglycerate kinase and a comparison with its mesophilic equivalent.	the structure of the phosphoglycerate kinase (pgk) from bacillus stearothermophilus, a moderate thermophile, has been determined and compared with that of its mesophilic equivalent from yeast. the bacillus enzyme structure was solved by molecular replacement and improved using constrained rigid-body, molecular dynamics and conventional refinement procedures. the refinement residual, calculated using all the measured data between 8 and 1.65 a, is 0.18(1). the stereo chemical deviations of the fin ...	1993	8456097
identification of cross-linked amino acids in the protein pair hmal23-hmal29 from the 50s ribosomal subunit of the archaebacterium haloarcula marismortui.	50s ribosomal subunits from the extreme halophilic archaebacterium haloarcula marismortui were treated with the homobifunctional protein-protein cross-linking reagents diepoxybutane (4 a) and dithiobis(succinimidyl propionate) (12 a). the dominant product with both cross-linking reagents was identified on the protein level as hmal23-hmal29, which is homologous to the protein pair l23-l29 from escherichia coli [walleczek, j., martin, t., redl, b., stöffler-meilicke, m., & stöffler, g. (1989) bioc ...	1993	8457554
structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase from trypanosoma brucei determined from laue data.	the three-dimensional structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase [d-glyceraldehyde-3-phosphate:nad+ oxidoreductase (phosphorylating), ec 1.12.1.12] from the sleeping-sickness parasite trypanosoma brucei was solved by molecular replacement at 3.2-a resolution with an x-ray data set collected by the laue method. for data collection, three crystals were exposed to the polychromatic synchrotron x-ray beam for a total of 20.5 sec. the structure was solved by using the bacillus s ...	1993	8460146
the energetics and cooperativity of protein folding: a simple experimental analysis based upon the solvation of internal residues.	the reversible unfolding of two dissimilar proteins, phosphoglycerate kinase from bacillus stearothermophilus (pgk) and staphylococcus aureus nuclease (san), was induced with two denaturants, urea and guanidinium chloride (guhcl). for each protein, structural transitions were monitored by intrinsic fluorescence intensity changes arising from a unique tryptophan residue. in the case of san the single, native tryptophan residue was used, whereas for pgk two versions, one with a tryptophan at posit ...	1993	8471598
primary structure, partial purification and regulation of key enzymes of the acetyl cycle of arginine biosynthesis in bacillus stearothermophilus: dual function of ornithine acetyltransferase.	a 3.4 kb ecori fragment, cloned in e. coli, that carries part of a cluster of genes encoding arginine biosynthetic functions of the thermophilic bacterium bacillus stearothermophilus, was sequenced on both strands. the sequence consists of a truncated argc gene, an argj region encoding a polypeptide with both n-acetylglutamate synthase and ornithine acetyltransferase activities, the argb gene and the n-terminal part of argd. the argb gene encodes a 258-amino-acid polypeptide with a deduced m(r) ...	1993	8473852