Publications

rapid, sensitive, microbial detection by gene amplification using restriction endonuclease target sequences.	the use of primers synthesized to eight class ii restriction endonuclease target sequences, from haemophilus parainfluenzae, escherichia coli, staphylococcus aureus, salmonella infantis, rhodobacter sphaeroides, klebsiella pneumoniae, bacillus amyloliquefaciens and proteus vulgaris for single and multiplex pcr identification of the organisms is discussed. results indicate that the method is sensitive and specific enough to detect single cells and attogram amounts of target dna. it has also been ...	1997	9281417
[riboflavin biosynthetic genes in bacillus amyloliquefaciens: primary structure, organization and regulation of activity].		1997	9297088
bacillus subtilis contains four closely related type i signal peptidases with overlapping substrate specificities. constitutive and temporally controlled expression of different sip genes.	most biological membranes contain one or two type i signal peptidases for the removal of signal peptides from secretory precursor proteins. in this respect, the gram-positive bacterium bacillus subtilis seems to be exceptional, because it contains at least four chromosomally-encoded type i signal peptidases, denoted sips, sipt, sipu, and sipv. here, we report the identification of the sipt and sipv genes, and the functional characterization of sipt, sipu, and sipv. the four signal peptidases hav ...	1997	9325333
dsc studies of the conformational stability of barstar wild-type.	the temperature induced unfolding of barstar wild-type of bacillus amyloliquefaciens (90 residues) has been characterized by differential scanning microcalorimetry. the process has been found to be reversible in the ph range from 6.4 to 8.3 in the absence of oxygen. it has been clearly shown by a ratio of delta hvh/delta hcal near 1 that denaturation follows a two-state mechanism. for comparison, the c82a mutant was also studied. this mutant exhibits similar reversibility, but has a slightly low ...	1997	9336842
isolation and purification of two novel streptomycete rnase inhibitors, sai14 and sai20, and cloning, sequencing, and expression in escherichia coli of the gene coding for sai14.	two new rnase inhibitors, sai14 (mr, approximately 14,000) and sai20 (mr, approximately 20,000), were isolated and purified from a streptomyces aureofaciens strain. the gene sai14, coding for sai14 protein, was cloned and expressed in escherichia coli. the alignment of the deduced amino acid sequence of sai14 with that of barstar, the rnase inhibitor from bacillus amyloliquefaciens, showed significant similarity between them, especially in the region which contains most of the residues involved ...	1998	9515932
enzymatic properties of a novel liquefying alpha-amylase from an alkaliphilic bacillus isolate and entire nucleotide and amino acid sequences.	a novel liquefying alpha-amylase (lamy) was found in cultures of an alkaliphilic bacillus isolate, ksm-1378. the specific activity of purified lamy was approximately 5,000 u mg of protein-1, a value two- to fivefold greater between ph 5 and 10 than that of an industrial, thermostable bacillus licheniformis enzyme. the enzyme had a ph optimum of 8.0 to 8.5 and displayed maximum activity at 55 degreesc. the molecular mass deduced from sodium dodecyl sulfate-polyacrylamide gel electrophoresis was a ...	1998	9726872
crystallization and preliminary x-ray investigation of the complex of rnase sa with wild-type barstar.	rnase sa, an extracellular ribonuclease produced by streptomyces aureofaciens, is inhibited by barstar, the natural protein inhibitor of barnase, the ribonuclease of bacillus amyloliquefaciens. the complex of rnase sa with wild-type barstar was crystallized by hanging-drop vapour diffusion. it was shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis that rnase sa and barstar are present in equimolar proportions in the crystals. the crystals are in the hexagonal space group p65 with ...	1998	9761909
conformational analysis of gpa and gpap in aqueous solution by molecular dynamics and statistical methods.	barnase, an extracellular endoribonuclease from bacillus amyloliquefaciens, hydrolyses single-stranded rna. its very low catalytic activity toward gpn dinucleotides, where n stands for any nucleoside, is markedly increased when a phosphate is added to the 3'-end, as in gpnp. here we investigate the conformational properties of gpa and gpap in solution, in order to determine whether differences in these properties may be related to the changes in enzymatic activity. two independent 1.3 ns molecul ...	1998	9790845
from detergent additive to semisynthetic peroxidase-simplified and up-scaled synthesis of seleno-subtilisin	a simplified and up-scaled synthesis of the semisynthetic peroxidase seleno-subtilisin was developed. highly purified to technical grade subtilisin preparations from bacillus licheniformis and bacillus amyloliquefaciens were applied as starting materials. activation of ser 221 with phenylmethanesulfonyl fluoride, nucleophilic substitution by sodium hydrogen selenide, and oxidation to the seleninic acid with hydrogen peroxide completed the chemical active-site modification. the reactions were acc ...	1998	10099399
the unusually slow unfolding rate causes the high stability of pyrrolidone carboxyl peptidase from a hyperthermophile, pyrococcus furiosus: equilibrium and kinetic studies of guanidine hydrochloride-induced unfolding and refolding.	to elucidate the energetic features of the anomalously high-level stabilization of a hyperthermophile pyrrolidone carboxyl peptidase (pfpcp) from a hyperthermophilic archaeon, pyrococcus furiosus, equilibrium and kinetic studies of the guanidine hydrochloride (guhcl)-induced unfolding and refolding were carried out with cd measurements at 220 nm in comparison with those from the mesophile homologue (bapcp) from bacillus amyloliquefaciens. the mutant protein of pfpcp substituted with ser at both ...	1998	9860869
discrepancies between the nmr and x-ray structures of uncomplexed barstar: analysis suggests that packing densities of protein structures determined by nmr are unreliable.	the crystal structure of the c82a mutant of barstar, the intracellular inhibitor of the bacillus amyloliquefaciens ribonuclease barnase, has been solved to a resolution of 2.8 a. the molecule crystallizes in the space group i41 with a dimer in the asymmetric unit. an identical barstar dimer is also found in the crystal structure of the barnase-barstar complex. this structure of uncomplexed barstar is compared to the structure of barstar bound to barnase and also to the structure of barstar solve ...	1998	9578582
recognition of rnase sa by the inhibitor barstar: structure of the complex at 1.7 a resolution.	we report the 1.7 a resolution structure of rnase sa complexed with the polypeptide inhibitor barstar. the crystals are in the hexagonal space group p65 with unit-cell dimensions a = b = 56.9, c = 135.8 a and the asymmetric unit contains one molecule of the complex. rnase sa is an extracellular microbial ribonuclease produced by streptomyces aureofaciens. barstar is the natural inhibitor of barnase, the ribonuclease of bacillus amyloliquefaciens. it inhibits rnase sa and barnase in a similar man ...	1998	9757110
thermodynamic stability of a cold-active alpha-amylase from the antarctic bacterium alteromonas haloplanctis.	the thermal stability of the cold-active alpha-amylase (aha) secreted by the antarctic bacterium alteromonas haloplanctis has been investigated by intrinsic fluorescence, circular dichroism, and differential scanning calorimetry. it was found that this heat-labile enzyme is the largest known multidomain protein exhibiting a reversible two-state unfolding, as demonstrated by the recovery of deltahcal values after consecutive calorimetric transitions, a deltahcal/deltaheff ratio close to unity, an ...	1999	10194383
effect of temperature on the saccharide composition obtained after alpha-amylolysis of starch	the hydrolysis of starch to low-molecular-weight products (normally characterised by their dextrose equivalent (de), which is directly related to the number-average molecular mass) was studied at different temperatures. amylopectin potato starch, lacking amylose, was selected because of its low tendency towards retrogradation at lower temperatures. bacillus licheniformis alpha-amylase was added to 10% [w/w] gelatinised starch solutions. the hydrolysis experiments were done at 50, 70, and 90 degr ...	1999	10099614
cloning and expression of a gene encoding the lytic functions of bacillus amyloliquefaciens phage: evidence of an auxiliary lysis system.	a bacteriophage specific to bacillus amyloliquefaciens, a gram-positive bacterium, was isolated from a local sewage treatment center. using a lysis assay, a gene, lys1521, was isolated and its nucleotide sequence revealed one open reading frame of 375 bp. homology studies showed amino acid alignment similarity with gene 5a of bacillus subtilis phages pza and phi29. overexpression of the cloned gene yielded a 13 kda protein corresponding to the predicted gene product. despite the fact that no sig ...	1999	16232602
effect of hypergravitational stress on microbial cell viability.	cell of escherichia coli b, thiobacillus intermedius 13-1, bacillus amyloliquefaciens ifo14141, staphylococcus aureus iid975, and saccharomyces cerevisiae kyokai no. 7 cultivated in nutrient media were subjected to hypergravitational stress for a period of between 1 and 24 h at 450,000 x g. the e. coli, b. amyloliquefaciens and s. cerevisiae cells showed survival rates of 38.5%, 0.005%, and 14.7%, respectively, after gravity treatment for 24 h as determined by their ability for colony formation, ...	1999	16232625
the crystal structure of pyroglutamyl peptidase i from bacillus amyloliquefaciens reveals a new structure for a cysteine protease.	the n-terminal pyroglutamyl (pglu) residue of peptide hormones, such as thyrotropin-releasing hormone (trh) and luteinizing hormone releasing hormone (lh-rh), confers resistance to proteolysis by conventional aminopeptidases. specialized pyroglutamyl peptidases (pgps) are able to cleave an n-terminal pyroglutamyl residue and thus control hormonal signals. until now, no direct or homology-based three-dimensional structure was available for any pgp.	1999	10196127
structural details of urea binding to barnase: a molecular dynamics analysis.	the molecular mechanism of urea-induced protein unfolding has not been established. it is generally thought that denaturation results from the stabilizing interactions of urea with portions of the protein that are buried in the native state and become exposed upon unfolding of the protein.	1999	10378267
a new phagemid vector for positive selection of recombinants based on a conditionally lethal barnase gene.	a new phagemid cloning vector for positive selection of recombinants, pba-7, was constructed which contains an active barnase gene encoding the cytotoxic ribonuclease from bacillus amyloliquefaciens, under control of the lac promoter. pba-7 is a derivative of the high-copy number pbluescript ii ks+ phagemid in which the modified barnase killer gene has been fused downstream from the lac promoter of the pbluescript ii ks+ multiple restriction site. when a laciq-negative escherichia coli strain is ...	1999	10386620
comparative enzymatic hydrolysis of phytate in various animal feedstuff with two different phytases.	bacillus amyloliquefaciens ds11 phytase (ds11 phytase) and aspergillus ficuum phytase (af phytase) activities were investigated by measuring the release of phosphate from phytate in animal feedstuff such as wheat bran, corn meal, soybean meal and rice flour at ph 5 and 7. in all the tested feedstuff, the enzymatic activity of ds11 phytase was more active at ph 7, but that of af phytase was more active at ph 5. from these results, the phytate in the gastrointestinal tract could be degraded in the ...	1999	10593587
refinement and structural analysis of barnase at 1.5 a resolution.	the structure of bacillus amyloliquefaciens ribonuclease (barnase), an extracellular 110-residue enzyme initially solved at 2.0 a resolution, has been refined at 1.5 a using synchrotron radiation and an imaging-plate scanner. refinement with anisotropic atomic displacement parameters resulted in increased accuracy of the structure. the final model has a crystallographic r factor of 11.5% and an rfree of 17.4%. the three independent molecules in the asymmetric unit, referred to as a, b and c, all ...	1999	10089345
preliminary x-ray crystallographic analysis of a novel phytase from a bacillus amyloliquefaciens strain.	a novel bacterial phytase from a bacillus amyloliquefaciens strain was crystallized using the hanging-drop vapour-diffusion method. the amino-acid sequence of the enzyme does not show any homology to those of other known phytases or phosphatases, with the exception of a phytase from bacillus subtilis. the enzyme exhibits a thermal stability which is strongly dependent on calcium ions. high-quality single crystals of the enzyme in the absence of calcium ions were obtained using a precipitant solu ...	1999	10089471
high-level expression of a recombinant thermostable phytase in bacillus subtilis.	an efficient expression system was developed in bacillus subtilis for the large scale production of phytase. the phytase gene with a native promoter derived from bacillus amyloliquefaciens was cloned in the bacillus expression vector pjh27 under a strong bj27 promoter and its expression was optimized. the expression of the phytase gene occurred during late exponential growth and the extracellular phytase production was 2.0 units/ml, which constituted over 90% of the total protein. the yield was ...	1999	27373920
detection of the dipicolinic acid biomarker in bacillus spores using curie-point pyrolysis mass spectrometry and fourier transform infrared spectroscopy.	thirty-six strains of aerobic endospore-forming bacteria confirmed by polyphasic taxonomic methods to belong to bacillus amyloliquefaciens, bacillus cereus, bacillus licheniformis, bacillus megaterium, bacillus subtilis (including bacillus niger and bacillus globigii), bacillus sphaericus, and brevi laterosporus were grown axenically on nutrient agar, and vegetative and sporulated biomasses were analyzed by curie-point pyrolysis mass spectrometry (pyms) and diffuse reflectance-absorbance fourier ...	2000	10655643
interspecific transformation of bacillus subtilis competent cells by chromosomal dna in lysates of protoplasts of bacillus amyloliquefaciens.	competent cells of bacillus subtilis were transformed with chromosomal dna in lysates of protoplasts of b. subtilis or b. amyloliquefaciens. the interspecific transformation frequency of b. subtilis by cysa in a conserved region was 3.1 x 10(4) transformants per microg dna, 60 times higher than that for conventional transformation using purified dna. increased interspecific transformation frequencies of b. subtilis were also observed for arg-1, lys-1, leub, arog, thr-5, hish, or metc markers out ...	2000	10737181
structural analysis of a chimeric bacterial alpha-amylase. high-resolution analysis of native and ligand complexes.	several chimeric alpha-amylases genes were constructed by an in vivo recombination technique from the bacillus amyloliquefaciens and bacillus licheniformis genes. one of the fusion amylases (hereafter ba2), consisting of residues 1-300 from b. amyloliquefaciens and 301-483 from b. licheniformis, has been extensively studied by x-ray crystallography at resolutions between 2.2 and 1.7 a. the 3-dimensional structure of the native enzyme was solved by multiple isomorphous replacement, and refined at ...	2000	10924103
purification and characterization of a new xylanase from acrophialophora nainiana.	a new xylanase activity (xynii) was isolated from liquid state cultures of acrophialophora nainiana containing birchwood xylan as carbon source. xynii was purified to apparent homogeneity by gel filtration and ion exchange chromatographies. the enzyme was optimally active at 55 degrees c and ph 7.0. xynii had molecular mass of 22630+/-3.0 and 22165 da, as determined by mass spectrometry and sds-page, respectively. the purified enzyme was able to act only on xylan as substrate. the apparent k(m) ...	2000	10989179
phylogenetic analysis of bacillus subtilis and related taxa based on partial gyra gene sequences.	partial gyra sequences were determined for twelve strains belonging to bacillus amyloliquefaciens, b. atrophaeus, b. licheniformis, b. mojavensis, b. subtilis subsp. subtilis, b. subtilis subsp. spizizenii and b. vallismortis. the average nucleotide and translated amino acid similarities for the seven type strains were 83.7 and 95.1%, respectively, whereas the corresponding value for the 16s rrna sequences was 99.1%. all of the type strains were sharply separated; the closest relationship was fo ...	2000	11204764
purification and some properties of a novel chitosanase from bacillus subtilis kh1.	one of at least two chitosanases secreted in the culture filtrate of bacillus subtilis kh1 was purified by two sequential deae sepharose cl-6b chromatographies, followed by sephacryl s-100 hr gel chromatography. the purified enzyme was homogenous as judged by sds-page. it showed an estimated molecular weight and pi of 28,000 and 8.3, respectively. the enzyme drastically reduced the viscosity of highly deacetylated chitosan substrates, with the subsequent formation of chitooligosaccharides [(glcn ...	2000	12483600
analysis of myo-inositol hexakisphosphate hydrolysis by bacillus phytase: indication of a novel reaction mechanism.	phytic acid (myo-inositol hexakisphosphate, insp(6)) hydrolysis by bacillus phytase (phyc) was studied. the enzyme hydrolyses only three phosphates from phytic acid. moreover, the enzyme seems to prefer the hydrolysis of every second phosphate over that of adjacent ones. furthermore, it is very likely that the enzyme has two alternative pathways for the hydrolysis of phytic acid, resulting in two different myo-inositol trisphosphate end products: ins(2,4,6)p(3) and ins(1,3,5)p(3). these results, ...	2000	11104666
[mald-ms in the quantitative analysis of peptides and proteins].	a modified method of isotope dilution was applied to the quantitative determination of peptides and proteins by maldi ms at subpicomolar level. the essence of the method consists in the quantitative analysis of the enzymic hydrolysis products rather than the starting compounds. this allows the measurements to be performed at a higher resolution and makes the method independent of the molecular mass of oligopeptides and proteins examined. fragments obtained by hydrolysis of the same oligopeptide ...	2000	11036525
improving the activity of immobilized subtilisin by site-directed attachment through a genetically engineered affinity tag.	an octapeptide affinity tag, asp-tyr-lys-asp-asp-asp-asp-lys (temied flag), was genetically fused to the c-terminus of subtilisin bpn' (sbt) from bacillus amyloliquefaciens. the fusion protein sbt-flag was immobilized to nonporous polystyrene and silica beads both in a site-directed and a random fashion. site-directed immobilization was achieved by employing the interaction between protein a and a monoclonal antibody specific for the flag peptide, while random immobilization was obtained by usin ...	2001	11293705
[dna-(n4-cytosine)-methyltransferase from bacillus amyloliquefaciens: kinetic and substrate binding properties].	interaction of dna-(n4-cytosine)-methyltransferase from the bacillus amyloliquefaciens (bamhi mtase, 49 kda) with a 20-mer oligonucleotide duplex containing the palindrome recognition site ggatcc was studied by methods of steady-state and presteady-state kinetics of the methyl group transfer, gel retardation, and crosslinking of the enzyme subunits with glutaric aldehyde. in steady-state conditions, bamhi mtase displays a simple kinetic behavior toward a 20-mer oligonucleotide substrate. a linea ...	2001	11234382
conformational characterization of designed minibarnase.	we have designed a minibarnase by removing one module from barnase, a bacterial rnase from bacillus amyloliquefaciens. barnase, consisting of 110 amino acid residues, is decomposed into six modules, m1-m6. module is defined as a peptide segment consisting of contiguous amino acid residues that makes a small compact conformation within a globular domain. to understand the role of module in protein architecture, we analyzed nmr and cd spectra of a minibarnase, which lacked 26 amino acid residues c ...	2001	11169386
the mechanism of aubstrate eecognition of pyroglutamyl-peptidase i from bacillus amyloliquefaciens as determined by x-ray crystallography and site-directed mutagenesis.	pyroglutamyl-peptidase is able to specifically remove the amino-terminal pyroglutamyl residue protecting proteins or peptides from aminopeptidases. to clarify the mechanism of substrate recognition for the unique structure of the pyrrolidone ring, x-ray crystallography and site-directed mutagenesis were applied. the crystal structure of pyroglutamyl-peptidase bound to a transition state analog inhibitor (inh), pyroglutaminal, was determined. two hydrogen bonds were located between the main chain ...	2001	11359794
conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells.	conditional expression of suicide genes in vivo has a wide range of applications in biological research and requires a minimal basal promoter activity in the uninduced state. to reduce basal activity of tetracycline (tc)-inducible target promoters we combined synthetic tet operators in varying numbers with a core promoter derived from the plant viral 35s promoter. an optimized promoter, p(tf), was found to exert a stringent regulation of luciferase in combination with tta and rtta in different m ...	2001	11504884
enhancement of the thermal stability of pyroglutamyl peptidase i by introduction of an intersubunit disulfide bond.	from the comparison of the three-dimensional structure of mesophilic pyroglutamyl peptidase from bacillus amyloliquefaciens and the thermophilic enzyme from thermococcus litoralis, the intersubunit disulfide bond was estimated to be one of the factors for thermal stability. since ser185 was corresponded to cys190 of the thermophilic enzyme by sequence alignment, the ser185 residue was replaced with cysteine by site-directed mutagenesis. the s185c mutant enzyme appeared to form a disulfide bond, ...	2001	11410277
thermal stability of pyrrolidone carboxyl peptidases from the hyperthermophilic archaeon, pyrococcus furiosus.	the temperature adaptation of pyrrolidone carboxyl peptidase (pcp) from a hyperthermophile, pyrococcus furiosus (pf pcp), was characterized in the context of an assembly form of the protein which is a homotetramer at neutral ph. the pf pcp exhibited maximal catalytic activity at 90-95 degrees c and its activity was higher in the temperature range 30-100 degrees c than its counterpart from the mesophilic bacillus amyloliquefaciens (bapcp). thermal stability was monitored by differential scanning ...	2001	11389725
one-step purification of glucoamylase by affinity precipitation with alginate.	it was found that alginate binds to glucoamylase, presumably through the recognition of starch binding domain of the latter. the present work exploits this for purification of glucoamylases from commercial preparation of aspergillus niger and crude culture filtrate of bacillus amyloliquefaciens by affinity precipitation technique in a single-step protocol. glucoamylase is selectively precipitated using alginate as macroaffinity ligand and later eluted with 1.0 m maltose. in the case of a. niger, ...	2001	11746949
molecular cloning and characterization of mycobacterium bovis bcg pcp gene encoding pyrrolidone carboxyl peptidase.	the mycobacterium bovis bacilli calmette-guerin (bcg) pcp gene that encodes the pyrrolidone carboxyl peptidase (pcp) was cloned from a lambdagtll genomic library and sequenced. the nucleotide sequence contains a 669 bp open reading frame coding for a protein of 222 amino acid residues with a calculated molecular mass of 23,209 da. the deduced amino acid sequence is highly homologous to the pcps from bacillus amyloliquefaciens, pseudomonas fluorescens, bacillus subtilis, streptococcus pyogenes, a ...	2001	11804334
quantitation of the capacity of the secretion apparatus and requirement for prsa in growth and secretion of alpha-amylase in bacillus subtilis.	regulated expression of amyq alpha-amylase of bacillus amyloliquefaciens was used to examine the capacity of the protein secretion apparatus of b. subtilis. one b. subtilis cell was found to secrete maximally 10 fg of amyq per h. the signal peptidase sipt limits the rate of processing of the signal peptide. another limit is set by prsa lipoprotein. the wild-type level of prsa was found to be 2 x 10(4) molecules per cell. decreasing the cellular level of prsa did not decrease the capacity of the ...	2001	11222585
rapid detection and identification of bacterial strains by fourier transform near-infrared spectroscopy.	the use of fourier transform near-infrared (ft-nir) spectroscopy and multivariate pattern recognition techniques for the rapid detection and identification of bacterial contamination in liquids was evaluated. the complex biochemical composition of bacteria yields ft-nir vibrational transitions (overtone and combination bands) that can be used for classification and identification. bacterial suspensions (escherichia coli hb101, e. coli atcc 43888, e. coli 1224, bacillus amyloliquifaciens, pseudom ...	2001	11261995
chemical modification of lysine residues in bacillus alpha-amylases: effect on activity and stability.	chemical modification of lysine residues in two bacterial alpha-amylases, a mesophilic enzyme from bacillus amyloliquefaciens (baa) and a thermophilic enzyme from bacillus licheniformis (bla) was carried out using citraconic anhydride. 13 +/- 1 residues in baa and 10 +/- 1 residues in bla were found modified under defined experimental conditions. modification brought about dramatic enhancement of thermal stability of baa and catalytic activity of bla. such alterations were found dependent on the ...	2001	11267650
detergent-independent in vitro activity of a truncated bacillus signal peptidase.	the gram-positive eubacterium bacillus subtilis contains five chromosomally encoded type i signal peptidases (spases) for the processing of secretory pre-proteins. even though four of these spases, denoted sips, sipt, sipu and sipv, are homologous to the unique spase i of escherichia coli, they are structurally different from that enzyme, being almost half the size and containing one membrane anchor instead of two. to investigate whether the unique membrane anchor of bacillus spases is required ...	2001	11283286
a bacillus amyloliquefaciens chbb protein binds beta- and alpha-chitin and has homologues in related strains.	a small (19.8 kda) protein was identified in bacillus amyloliquefaciens alko 2718 cultures during growth in the presence of yeast extract and chitin, but not with glucose. the protein targets beta-chitin best, then alpha-chitin, but barely any other polysaccharide. this described chitin-binding protein (chbb) is the first of its type from a bacillus strain and cross-reacts with antibodies raised against the streptomyces alpha-chitin-binding protein chb1. using reverse genetics, the chromosomal c ...	2001	11429457
x-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, pyrococcus furiosus, and its cys-free mutant.	in order to elucidate the mechanism of the thermostability of proteins from hyperthermophiles, x-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, pyrococcus furiosus (pfpcp), and its mutant protein with ser substituted at cys142 and cys188 were determined at 2.2 and 2.7 a resolution, respectively. the obtained structures were compared with those previously reported for pyrrolidone carboxyl peptidases from a hyperthermophilie, thermococcus litoralis (tlpcp), a ...	2001	11432786
functional analysis of antibacterial activity of bacillus amyloliquefaciens phage endolysin against gram-negative bacteria.	to analyze the antibacterial activity of bacillus amyloliquefaciens phage endolysin, nine deletion derivatives of the endolysin were constructed. each deletion mutant was overexpressed, purified and characterized. the catalytic domain was located on the n-terminal region and the c-terminus had an affinity with the bacterial envelope. the enzymatic activity remained in spite of the deletion of the c-terminal 116-amino acid region; however, the antibacterial activity was lost. these results indica ...	2001	11434926
deduced amino-acid sequence of a calcium-free alpha-amylase from a strain of bacillus: implications from molecular modeling of high oxidation stability and chelator resistance of the enzyme.	alkaline alpha-amylase (amyk38) from the alkaliphilic bacillus sp. strain ksm-k38 is a unique enzyme in that it is highly chelator-resistant and oxidatively stable [hagihara, h., igarashi, k., hayashi, y., endo, k., ikawa-kitayama, k., ozaki, k., kawai, s. & ito, s. (2001) appl. environ. microbiol. 67, 1744-1750]. this enzyme was found to contain no ca and require na (or monovalent cations) for manifestation of activity. the nucleotide sequence of the gene for the novel enzyme was determined, an ...	2001	11453991
proteolysis of mesophilic and thermophilic alpha-amylases: a comparative study.	a comparative study was performed on limited and extensive proteolysis of mesophilic (from bacillus amyloliquefaciens [baa]) and thermophilic (from bacillus licheniformis [bla]) alpha-amylases using trypsin. as expected, the thermophilic enzyme showed greater resistance to digestion by the protease. while the catalytic potential of bla was enhanced on proteolysis, that of baa was diminished owing to this process. combined with greater catalytic activity, a lower thermal stability was observed fo ...	2001	11456297
free polymeric bioligands in aqueous two-phase affinity extractions of microbial xylanases and pullulanase.	two reversibly soluble-insoluble polymers (viz. eudragit s-100 and alginate) were used as free macroaffinity bioligands in polyethylene glycol (peg)/salt two-phase systems for separation of enzymes. incorporation of eudragit s-100 and alginate in the peg phase led to considerable selectivity in separation of microbial xylanases and pullulanase, respectively. xylanase from aspergillus niger was recovered 93% with 56-fold purification, whereas the enzyme from trichoderma reesei and bacillus amylol ...	2001	11483013
activity and stability of a thermostable alpha-amylase compared to its mesophilic homologue: mechanisms of thermal adaptation.	to elucidate how enzymes adapt to extreme environmental conditions, a comparative study with a thermostable alpha-amylase from bacillus licheniformis (bla) and its mesophilic homologue from bacillus amyloliquefaciens (baa) was performed. we measured conformational stability, catalytic activity, and conformational fluctuations on the picosecond time scale for both enzymes as a function of temperature. the objective of this study is to analyze how these properties are related to each other. bla sh ...	2001	11524019
calcium-dependent catalytic activity of a novel phytase from bacillus amyloliquefaciens ds11.	the thermostable phytase from bacillus amyloliquefaciens ds11 hydrolyzes phytate (myo-inositol hexakisphosphate, ip6) to less phosphorylated myo-inositol phosphates in the presence of ca2+. in this report, we discuss the unique ca2+-dependent catalytic properties of the phytase and its specific substrate requirement. initial rate kinetic studies of the phytase indicate that the enzyme activity follows a rapid equilibrium ordered mechanism in which binding of ca2+ to the active site is necessary ...	2001	11583167
cloning and expression of an endocellulase gene from a novel streptomycete isolated from an east african soda lake.	alkaline cellulase-producing actinomycete strains were isolated from mud samples collected from east african soda lakes. the strains were identified as novel streptomyces spp. by 16s rdna sequence analysis. a cellulase gene (cel12a) from streptomyces sp. strain 11ag8 was cloned by expression screening of a genomic dna library in escherichia coli. from the nucleotide sequence of a 1.5-kb dna fragment, an open reading frame of 1,113 nucleotides was identified encoding a protein of 371 amino acids. ...	2001	11699647
antibacterial activity of bacillus amyloliquefaciens phage endolysin without holin conjugation.	to characterize the enzymatic activity and antibacterial activity of endolysin encoded by a bacillus amyloliquefaciens phage, the open reading frame encoding endolysin was amplified by pcr and cloned into the expression plasmid pet21d(+). the resultant plasmid was used to transform escherichia coli jm109(de3). production of endolysin in the cytosol facilitated cell lysis without coproduction of holin, which is considered to degrade or alter the cytoplasmic membrane. the phage endolysin was overe ...	2001	16233024
antimicrobial activity of culture filtrate of bacillus amyloliquefaciens rc-2 isolated from mulberry leaves.	abstract a potential antagonist, bacillus amyloliquefaciens strain rc-2, against colletotrichum dematium, mulberry anthracnose fungus, was obtained from healthy mulberry leaves by in vitro and in vivo screening techniques. application of culture filtrate of rc-2 inhibited disease on mulberry leaves, indicating that suppression was due to antifungal compounds in the filtrate. development of mulberry anthracnose on mulberry leaves was inhibited only when the culture filtrate was applied before fun ...	2001	18944392
directed evolution of barnase stability using proteolytic selection.	we report the construction of a phage-displayed repertoire of mutants of the ribonuclease barnase from bacillus amyloliquefaciens. the construction was guided by the natural variability between two closely related ribonucleases, barnase and binase from bacillus intermedius. this repertoire was selected using a proteolytic selection method, allowing sorting of the library according to the resistance of the mutants toward proteolysis. susceptibility toward proteolysis has been correlated with flex ...	2002	12368103
mulberry anthracnose antagonists (iturins) produced by bacillus amyloliquefaciens rc-2.	bacillus amyloliquefaciens strain rc-2 produced seven antifungal compounds (1-7) secreted into the culture filtrate. these compounds inhibited the development of mulberry anthracnose caused by the fungus, colletotrichum dematium. chemical structural analyses by nmr and fab-ms revealed that all these compounds were iturins (cyclic peptides with the following sequence: l-asn --> d-tyr --> d-asn --> l-gln --> l-pro --> d-asn --> l-ser --> d-beta-amino acid -->) and compounds 1-6 are identical to it ...	2002	12423891
extracellular phytase activity of bacillus amyloliquefaciens fzb45 contributes to its plant-growth-promoting effect.	several bacillus strains belonging to the b. subtilis/amyloliquefaciens group isolated from plant-pathogen-infested soil possess plant-growth-promoting activity [krebs, b. et al. (1998) j plant dis prot 105, 181-197]. three out of the four strains investigated were identified as b. amyloliquefaciens and were able to degrade extracellular phytate (myo-inositol hexakisphosphate). the highest extracellular phytase activity was detected in strain fzb45, and diluted culture filtrates of this strain s ...	2002	12101298
the x-ray crystal structure of pyrrolidone-carboxylate peptidase from hyperthermophilic archaea pyrococcus horikoshii.	the crystal structure of pyrrolidone-carboxylate peptidase (pcp) from hyperthermophilic archaea pyrococcus horikoshii (phopcp) has been determined at 1.6-a resolution by x-ray crystallography. pcp belongs to the c15 family of cysteine protease, and specifically removes the amino terminal pyroglutamate residue from a wide range of n-terminal-blocking peptides. the crystal structure is very similar to that of other hyperthermophiles, pyrococcus furiosus and thermococcus litoralis, and even that fr ...	2002	12836705
thermodynamic and activation parameters for the hydrolysis of amylose with bacillus alpha-amylases in a diluted anionic surfactant solution.	the effects of an anionic surfactant, sodium dodecyl sulfate (sds), on the kinetic behavior in the hydrolysis of amylose with two bacterial alpha-amylases from bacillus amyloliquefaciens and b. licheniformis were studied. the catalytic rates of both amylases in the present study showed sigmoidal kinetics with increased concentration of added sds; the rates were increased in the range below the critical micelle concentration (cmc) and then markedly decreased above the cmc. the catalytic rate of t ...	2002	16233236
bacillus amyloliquefaciens orthologue of bacillus subtilis ywro encodes a nitroreductase enzyme which activates the prodrug cb 1954.	a nitroreductase with distinct properties that can activate the prodrug 5-aziridinyl-2,4-dinitrobenzamide (cb 1954) was isolated from bacillus amyloliquefaciens. the encoding gene was identified as a homologue of the ywro of bacillus subtilis, and was obtained as a pcr product by reverse genetics, cloned and the entire nucleotide sequence determined. the gene was found to reside between homologues of the b. subtilis alsd and yswb genes; however, the ywro and yswb genes of b. amyloliquefaciens we ...	2002	11782522
bacillus endophyticus sp. nov., isolated from the inner tissues of cotton plants (gossypium sp.).	four strains of aerobic, endospore-forming bacteria were isolated from the inner tissues of healthy cotton plants (gossypium sp., dushanbe, tajikistan). the organisms had identical randomly amplified polymorphic dna patterns that distinguished them from other bacilli that are commonly isolated from plant tissues, e.g. bacillus amyloliquefaciens, bacillus licheniformis, bacillus pumilus and bacillus subtilis. pcr amplification of 16s-23s rrna spacer regions suggested that the four strains could b ...	2002	11837291
identification and properties of type i-signal peptidases of bacillus amyloliquefaciens.	the use of bacillus amyloliquefaciens for enzyme production and its exceptional high protein export capacity initiated this study where the presence and function of multiple type i signal peptidase isoforms was investigated. in addition to type i signal peptidases sips(ba) [meijer, w.j.j., de jong, a., bea, g., wisman, a., tjalsma, h., venema, g., bron, s. & van dijl, j.m. (1995) mol. microbiol. 17, 621-631] and sipt(ba) [hoang, v. & hofemeister, j. (1995) biochim. biophys. acta 1269, 64-68] whi ...	2002	11856304
purification and characterization of two antifungal chitinases extracellularly produced by bacillus amyloliquefaciens v656 in a shrimp and crab shell powder medium.	a gram-positive bacterium with antagonistic activity was isolated from the soil. it has been identified as bacillus amyloliquefaciens strain v656 on the basis of 16s ribosomal dna analysis and standard bacteriological tests. b. amyloliquefaciens v656 produced antifungal enzymes when it was grown in a medium containing shrimp and crab shell powder (scsp) of marine waste. the antifungal enzymes displayed chitinase activities. two extracellular antifungal chitinases (fi and fii) were purified and c ...	2002	11929278
construction of self-disruptive bacillus megaterium in response to substrate exhaustion for polyhydroxybutyrate production.	in order to establish a novel recovery system for polyhydroxyalkanoates, a self-disruptive strain of bacillus megaterium that responds to substrate exhaustion was constructed. a gene cassette carrying the lysis system of bacillus amyloliquefaciens phage - holin and endolysin - was inserted into the escherichia coli- bacillus subtilis shuttle vector px under the control of a xylose-inducible expression system, xylr-xyla '. in this system, the expression of a target gene is induced by xylose but i ...	2002	12111148
action pattern and subsite mapping of bacillus licheniformis alpha-amylase (bla) with modified maltooligosaccharide substrates.	this study represents the first characterisation of the substrate-binding site of bacillus licheniformis alpha-amylase (bla). it describes the first subsite map, namely, number of subsites, apparent subsite energies and the dual product specificity of bla. the product pattern and cleavage frequencies were determined by high-performance liquid chromatography, utilising a homologous series of chromophore-substituted maltooligosaccharides of degree of polymerisation 4-10 as model substrates. the bi ...	2002	11997021
rhizobacteria-induced resistance perturbs viral disease progress and triggers defense-related gene expression.	selected strain of nonpathogenic rhizobacterium extn-1 from the bacillus amyloliquefaciens is capable of eliciting broad-spectrum induced systemic resistance (isr) in several crops that is phenotypically similar to pathogen-induced systemic acquired resistance (sar). in tobacco (nicotiana tabacum cv. samsun-nn), extn-1 treatment also perturbs the disease progress by pepper mild mottle virus (pmmov), a member of tobamovirus group. to investigate the defense mechanisms induced by this rhizobacteri ...	2002	12019515
enterotoxin production in natural isolates of bacillaceae outside the bacillus cereus group.	thirty-nine bacillus strains obtained from a variety of environmental and food sources were screened by pcr for the presence of five gene targets (hblc, hbld, hbla, nhea, and nheb) in two enterotoxin operons (hbl and nhe) traditionally harbored by bacillus cereus. seven isolates exhibited a positive signal for at least three of the five possible targets, including bacillus amyloliquefaciens, b. cereus, bacillus circulans, bacillus lentimorbis, bacillus pasteurii, and bacillus thuringiensis subsp ...	2002	12039781
calcium-binding parameter of bacillus amyloliquefaciens alpha-amylase determined by inactivation kinetics.	the irreversible thermal inactivation and the thermodynamics of calcium ion binding of bacillus amyloliquefaciens alpha-amylase in the absence of substrates were studied. the enzyme inactivation on heating was apparently followed by first-order kinetics. the enzyme was stabilized with an increased concentration of calcium ion and thus the inactivation was highly dependent on the state of calcium binding. the activation parameter for the inactivation suggests an unfolding of the enzyme protein up ...	2002	12049626
reactions of alpha amylases with starch granules in aqueous suspension giving products in solution and in a minimum amount of water giving products inside the granule.	porcine pancreatic alpha amylase (ppa) and bacillus amyloliquefaciens alpha amylase (baa) were allowed to react with starch granules from maize, waxy maize, amylomaize-7, and potato in an aqueous suspension with a starch to water ratio of 1:10 and in a minimum of water with a starch to water ratio of 1:1. quantitative amounts of the maltodextrin products were determined by tlc and scanning densitometry. the two alpha amylases gave different products that were characteristic of their unique actio ...	2002	12062526
cytotoxic potential of industrial strains of bacillus sp.	the cytotoxic potential of selected strains of bacillus licheniformis, bacillus amyloliquefaciens, and bacillus subtilis, used in the production of industrial enzyme products, has been assessed. cytotoxicity was determined in chinese hamster ovary (cho-k1) cells by measuring total cellular metabolic activity using the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (mtt). initially the mtt assay was validated against toxigenic strains of bacillus cereus, to define t ...	2002	12460750
revision of the taxonomic position of the xylanolytic bacillus sp. mir32 reidentified as bacillus halodurans and plasmid-mediated transformation of b. halodurans.	bacillus sp. mir32 has been isolated using xylan as the only carbon source, and one of its xylanolytic enzymes has been extensively studied. biochemical analysis first related this strain to bacillus amyloliquefaciens, but further studies based on a comparison of 16s rdna sequences, g+c content, and dna-dna hybridization showed that strain mir32 should be classified as a member of the species bacillus halodurans. this change is also supported by the typical phenotype observed and by the results ...	2002	12382115
the pathway of dephosphorylation of myo-inositol hexakisphosphate by phytate-degrading enzymes of different bacillus spp.	the pathway of dephosphorylation of myo-inositol hexakisphosphate by the phytate-degrading enzymes of bacillus subtilis 168, bacillus amyloliquefaciens atcc 15841, and bacillus amyloliquefaciens 45 was established using a combination of high-performance ion chromatography analysis and kinetic studies. the data demonstrate that all the bacillus phytate-degrading enzymes under investigation dephosphorylate myo-inositol hexakisphosphate by sequential removal of phosphate groups via two independent ...	2002	12556126
cloning of the maltose phosphorylase gene from bacillus sp. strain rk-1 and efficient production of the cloned gene and the trehalose phosphorylase gene from bacillus stearothermophilus sk-1 in bacillus subtilis.	the maltose phosphorylase (mpase) gene of bacillus sp. strain rk-1 was cloned by pcr with oligonucleotide primers designed on the basis of a partial n-terminal amino acid sequence of the purified enzyme. the mpase gene consisted of 2,655 bp encoding a theoretical protein with a mr of 88,460, and had no secretion signal sequence, although most of the mpase activity was detected in the culture supernatant of rk-1. this cloned mpase gene and the trehalose phosphorylase (tpase) gene from bacillus st ...	2002	12596853
dna (cytosine-n4-)- and -(adenine-n6-)-methyltransferases have different kinetic mechanisms but the same reaction route. a comparison of m.bamhi and t4 dam.	we studied the kinetics of methyl group transfer by the bamhi dna-(cytosine-n(4)-)-methyltransferase (mtase) from bacillus amyloliquefaciens to a 20-mer oligodeoxynucleotide duplex containing the palindromic recognition site ggatcc. under steady state conditions the bamhi mtase displayed a simple kinetic behavior toward the 20-mer duplex. there was no apparent substrate inhibition at concentrations much higher than the k(m) for either dna (100-fold higher) or s-adenosyl-l-methionine (adomet) (20 ...	2003	12598537
purification and characterization of a fibrinolytic enzyme produced by bacillus amyloliquefaciens dc-4 screened from douchi, a traditional chinese soybean food.	bacillus amyloliquefaciens dc-4, which produces a strongly fibrinolytic enzyme, was isolated from douchi, a traditional chinese soybean-fermented food. a fibrinolytic enzyme (subtilisin dfe) was purified from the supernatant of b. amyloliquefaciens dc-4 culture broth and displayed thermophilic, hydrophilic and strong fibrinolytic activity. subtilisin dfe was demonstrated to be homogeneous by sds-page and isoelectric focusing electrophoresis, and has molecular mass of 28000 da and a pi of 8.0. th ...	2003	12524032
study of the inhibition of four alpha amylases by acarbose and its 4iv-alpha-maltohexaosyl and 4iv-alpha-maltododecaosyl analogues.	acarbose analogues, 4iv-maltohexaosyl acarbose (g6-aca) and 4iv-maltododecaosyl acarbose (g12-aca), were prepared by the reaction of cyclomaltodextrin glucanyltransferase with cyclomaltohexaose and acarbose. the inhibition kinetics of acarbose and the two acarbose analogues were studied for four different alpha-amylases: aspergillus oryzae, bacillus amyloliquefaciens, human salivary, and porcine pancreatic alpha-amylases. the three inhibitors showed mixed, noncompetitive inhibition, for all four ...	2003	14499573
production of chlamydia pneumoniae proteins in bacillus subtilis and their use in characterizing immune responses in the experimental infection model.	due to intracellular growth requirements, large-scale cultures of chlamydiae and purification of its proteins are difficult and laborious. to overcome these problems we produced chlamydial proteins in a heterologous host, bacillus subtilis, a gram-positive nonpathogenic bacterium. the genes of chlamydia pneumoniae major outer membrane protein (momp), the cysteine-rich outer membrane protein (omp2), and the heat shock protein (hsp60) were amplified by pcr, and the pcr products were cloned into ex ...	2003	12738633
phylogenetic relationships between bacillus species and related genera inferred from comparison of 3' end 16s rdna and 5' end 16s-23s its nucleotide sequences.	the nucleotide sequences of the 3' end of the 16s rdna and the 16s-23s internal transcribed spacer (its) of 40 bacillaceae species were determined. these included 21 bacillus, 9 paenibacillus, 6 brevibacillus, 2 geobacillus, 1 marinibacillus and 1 virgibacillus species. comparative sequence analysis of a 220 bp region covering a highly conserved 150 bp sequence located at the 3' end of the 16s rrna coding region and a conserved 70 bp sequence located at the 5' end of the 16s-23s its of the 40 sp ...	2003	12807189
similarities between the thermal inactivation kinetics of bacillus amyloliquefaciens alpha-amylase in an aqueous solution of sodium dodecyl sulphate and the kinetics in the solution of anionic-phospholipid vesicles.	an anionic surfactant, sds, is commonly used to model compounds of negatively charged phospholipids. the effect of sds on the thermal inactivation of baa (alpha-amylase from bacillus amyloliquefaciens ) was compared with the effect of negatively charged phospholipid vesicles of dopa (dioleoylphosphatidic acid) at 40-55 degrees c. the inactivation kinetics revealed that both sds below its c.m.c. (critical micellar concentration) and dopa vesicles accelerated the inactivation and the unfolding of ...	2003	12816534
the enzyme activity of a novel phytase from bacillus amyloliquefaciens ds11 and its potential use as a feed pellet.		2003	12833216
purification and characterization of proteases from bacillus amyloliquefaciens isolated from traditional soybean fermentation starter.	bacillus amyloliquefaciens fse-68 isolated from meju, a korean soybean fermentation starter, was identified on the basis of biophysical tests and 16s rrna gene sequence. a neutral metalloprotease (npr68) and an alkaline serine-protease (apr68) were purified by ammonium sulfate precipitation and cation exchange chromatography and identified on the basis of their activities at different ph values and the selective protease inhibitors. the molecular weights of npr68 and apr68 measured with esi-ms w ...	2003	14664526
magnetic alginate microparticles for purification of alpha-amylases.	spherical magnetic alginate microparticles (25-60 microm in diameter) were prepared using the microemulsion system, with water-saturated 1-pentanol as the organic phase. the limited solubility of 1-pentanol in water enabled simple removal of the organic solvent from the prepared beads with water solution. the prepared alginate microparticles were used as magnetic affinity adsorbents for specific purification of alpha-amylases. enzyme activity was eluted by 1.0 m maltose. alpha-amylases from baci ...	2003	14580797
real-time molecular beacon nasba reveals hblc expression from bacillus spp. in milk.	nucleic acid sequence-based amplification (nasba) was applied in combination with a fluorescein-conjugated molecular beacon specific for a sequence flanked by transcript-specific primers in order to monitor hblc enterotoxin gene expression in real-time from milk separately contaminated with bacillus amyloliquefaciens, bacillus cereus, and bacillus circulans. maximal enterotoxin expression was noted following 16, 15, and 16 h, respectively, when grown in artificially contaminated nonfat dried mil ...	2003	14592426
peptide ligands that bind selectively to spores of bacillus subtilis and closely related species.	as part of an effort to develop detectors for selected species of bacterial spores, we screened phage display peptide libraries for 7- and 12-mer peptides that bind tightly to spores of bacillus subtilis. all of the peptides isolated contained the sequence asn-his-phe-leu at the amino terminus and exhibited clear preferences for other amino acids, especially pro, at positions 5 to 7. we demonstrated that the sequence asn-his-phe-leu-pro (but not asn-his-phe-leu) was sufficient for tight spore bi ...	2003	14602648
effects of inactivation and constitutive expression of the unfolded- protein response pathway on protein production in the yeast saccharomyces cerevisiae.	one strategy to obtain better yields of secreted proteins has been overexpression of single endoplasmic reticulum-resident foldases or chaperones. we report here that manipulation of the unfolded-protein response (upr) pathway regulator, hac1, affects production of both native and foreign proteins in the yeast saccharomyces cerevisiae. the effects of hac1 deletion and overexpression on the production of a native protein, invertase, and two foreign proteins, bacillus amyloliquefaciens alpha-amyla ...	2003	12676684
macroaffinity ligand-facilitated three-phase partitioning (mlftpp) of alpha-amylases using a modified alginate.	the crude extracts of alpha-amylases when mixed with alginate, tert-butyl alcohol, and ammonium sulfate resulted in an interfacial precipitate containing polymer-bound amylase. the precipitate was dissolved in 1 m maltose to recover alpha-amylase activity. the recovery of alpha-amylases were 74%, 77%, and 92% in the case of bacillus amyloliquefaciens, wheat germ, and porcine pancreas, respectively. all purified preparations showed a single band on sds-page.	2003	12675592
studies of yeast kluyveromyces lactis mutations conferring super-secretion of recombinant proteins.	we have isolated mutants responsible for a super-secretion phenotype in kluyveromyces lactis using the gene coding for a bacillus amyloliquefaciens alpha-amylase as a marker for secretion. these mutations defined two groups, dominant and recessive. the recessive mutant strain, which secreted the heterologous protein in five-fold excess compared to the wild-type strain, was used for the cloning of genes, restraining the super-secreting phenotype. in screening for genes affecting super-secreting p ...	2003	12489121
[dna-[n4-cytosine]-methyltransferase from bacillus amyloliquefaciens: mechanism of action derived from steady state kinetics].	kinetic analysis of methyl group transfer from s-adenosyl-l-methionine (sam) to the 5'-ggatcc recognition site catalyzed by the dna-[n4-cytosine]-methyltransferase from bacillus amyloliquefaciens [ec 2.1.1.113] has shown that the dependence of the rate of methylation of the 20-meric substrate duplex on sam and dna concentration are normally hyperbolic, and the maximal rate is attained upon enzyme saturation with both substrates. no substrate inhibition is observed even at concentrations many tim ...	2003	12624955
pyroglutamyl-peptidase i: cloning, sequencing, and characterisation of the recombinant human enzyme.	pyroglutamyl-peptidase i (ec 3.4.19.3) is well known from bacteria and archaea, but has not previously been cloned or sequenced from any vertebrate. we describe the cloning and sequencing of the human (aj278828) and mouse (aj278829) forms of pyroglutamyl-peptidase i. the deduced amino acid sequences each consist of 209 residues and show approximately 30% identity with bacterial forms of the enzyme. they show clear homology to the enzyme from prokaryotes and place the mammalian forms of the enzym ...	2003	12651114
monte carlo simulation of the alpha-amylolysis of amylopectin potato starch. 2. alpha-amylolysis of amylopectin.	a model is presented that describes all the saccharides that are produced during the hydrolysis of starch by an alpha-amylase. potato amylopectin, the substrate of the hydrolysis reaction, was modeled in a computer matrix. the four different subsite maps presented in literature for alpha-amylase originating from bacillus amyloliquefaciens were used to describe the hydrolysis reaction in a monte carlo simulation. the saccharide composition predicted by the model was evaluated with experimental va ...	2003	14618387
directed evolution of a bacterial alpha-amylase: toward enhanced ph-performance and higher specific activity.	alpha-amylases, in particular, microbial alpha-amylases, are widely used in industrial processes such as starch liquefaction and pulp processes, and more recently in detergency. due to the need for alpha-amylases with high specific activity and activity at alkaline ph, which are critical parameters, for example, for the use in detergents, we have enhanced the alpha-amylase from bacillus amyloliquefaciens (baa). the genes coding for the wild-type baa and the mutants baa s201n and baa n297d were s ...	2003	14500872
secondary calcium-binding parameter of bacillus amyloliquefaciens alpha-amylase obtained from inhibition kinetics.	calcium is required for the stabilization of alpha-amylase because of primary binding (essential binding), but has been shown to inhibit hydrolytic catalysis due to secondary binding at the catalytic site in the enzyme. the role of calcium in the hydrolysis of soluble amylose by bacillus amyloliquefaciens alpha-amylase was characterized using the equilibrium dissociation constant (k(m)) and k(cat) for the hydrolytic catalysis. the enzymatic hydrolysis was inhibited by a relatively high concentra ...	2003	16233519
[i87e mutation prevents barstar dimerization].	the c40,82a;i87e mutant of barstar, an intracellular inhibitor of the ribonuclease barnase from bacillus amyloliquefaciens, was obtained, and its physicochemical properties were studied. it was produced as a fusion protein with thioredoxin and then cleaved from this by ekmax enterokinase. the mutant was shown by nmr to retain the spatial structure of the wild-type protein but, in contrast to barstar, does not form the homodimers characteristic of barstar in aqueous solution. the mutant protein b ...	2004	15586816
taxonomic characterization and plant colonizing abilities of some bacteria related to bacillus amyloliquefaciens and bacillus subtilis.	the phylogenetic relationships of 17 bacillus strains isolated from plants and soil were determined from partial sequences of genes encoding 16s rrna, gyrasea (gyra) and the chea histidine kinase. five strains were closely related to bacillus subtilis subsp. subtilis, three strains were more closely related to b. subtilis subsp. spizizeni and two strains were identified as b. mojavensis. the remaining seven strains formed a cluster closely related to, but distinct from, bacillus amyloliquefacien ...	2004	19712408
bacillus amyloliquefaciens strains isolated from moisture-damaged buildings produced surfactin and a substance toxic to mammalian cells.	fungicidic bacillus amyloliquefaciens strains isolated from the indoor environment of moisture-damaged buildings contained heat-stable, methanol-soluble substances that inhibited motility of boar spermatozoa within 15 min of exposure and killed feline lung cells in high dilution in 1 day. boar sperm cells lost motility, cellular atp, and nadh upon contact to the bacterial extract (0.2 microg dry wt/ml). two bioactive substances were purified from biomass of the fungicidal isolates. one partially ...	2004	15014930
production of an antifungal protein for control of colletotrichum lagenarium by bacillus amyloliquefaciens met0908.	a plant pathogenic fungus, colletotrichum lagenarium, causing watermelon anthracnose, was isolated from naturally infected leaves, stems, and fruits of watermelon. a bacterial strain, met0908, showing a potent antifungal activity against c. lagenarium, was isolated from soil. an antifungal protein was purified by 30% ammonium sulfate saturation and concentrated using centricon 10, deae-sepharose(tm) fast flow column and sephacryl s-100 gel filtration chromatography. the molecular weight of the p ...	2004	15109737
stability parameters for one-step mechanism of irreversible protein denaturation: a method based on nonlinear regression of calorimetric peaks with nonzero deltacp.	thermal transitions of many proteins have been found to be calorimetrically irreversible and scan-rate dependent. calorimetric determinations of stability parameters of proteins which unfold irreversibly according to a first-order kinetic scheme have been reported. these methods require the approximation that the increase in heat capacity upon denaturation deltacp is zero. a method to obtain thermodynamic parameters and activation energy for the two-state irreversible process n --> d from nonlin ...	2004	15113687
isolation, characterization, and identification of bacterial contaminants in semifinal gelatin extracts.	bacterial contamination of gelatin is of great concern. indeed, this animal colloid has many industrial applications, mainly in food and pharmaceutical products. in a previous study (e. de clerck and p. de vos, syst. appl. microbiol. 25:611-618), contamination of a gelatin production process with a variety of gram-positive and gram-negative bacteria was demonstrated. in this study, bacterial contamination of semifinal gelatin extracts from several production plants was examined. since these extr ...	2004	15184171
comparative studies on trifluoroethanol (tfe) state of a thermophilic alpha-amylase and its mesophilic counterpart: limited proteolysis, conformational analysis, aggregation and reactivation of the enzymes.	detailed circular dichroism (cd), scattering and quenching studies, 1-anilinonaphthalene-8-sulfonate (ans) binding, irreversible thermoinactivation, activity measurements and proteolytic digestion of bacterial alpha-amylases have been carried out to elucidate the effect of trifluoroethanol (tfe) on the structure of these enzymes. under high concentrations of tfe both of the alpha-amylases, a thermostable alpha-amylase from bacillus licheniformis (bla) and its mesophilic counterpart from bacillus ...	2004	15225989