| the crystal structure of the bacillus lentus alkaline protease, subtilisin bl, at 1.4 a resolution. | the crystal structure of subtilisin bl, an alkaline protease from bacillus lentus with activity at ph 11, has been determined to 1.4 a resolution. the structure was solved by molecular replacement starting with the 2.1 a structure of subtilisin bpn' followed by molecular dynamics refinement using x-plor. a final crystallographic r-factor of 19% overall was obtained. the enzyme possesses stability at high ph, which is a result of the high pi of the protein. almost all of the acidic side-chains ar ... | 1992 | 1453465 |
| introduction of a free cysteinyl residue at position 68 in the subtilisin savinase, based on homology with proteinase k. | two subfamilies of the subtilisins, distinguished by the presence or absence of a free cysteinyl residue near the essential histidyl residue of the catalytic triad, are known. in order to evaluate the significance of the presence of this -sh group a cysteinyl residue has been introduced by site-directed mutagenesis into the cysteine-free subtilisin-like enzyme from bacillus lentus, i.e. savinase. the free cysteine affects the enzyme activity only slightly but renders it sensitive to mercurials p ... | 1992 | 1551423 |
| extensive comparison of the substrate preferences of two subtilisins as determined with peptide substrates which are based on the principle of intramolecular quenching. | subtilisins are serine endopeptidases with an extended binding cleft comprising at least eight binding subsites. interestingly, subsites distant from the scissile bond play a dominant role in determining the specificity of the enzymes. the development of internally quenched fluorogenic substrates, which allow polypeptides of more than 11 amino acids to be inserted between the donor and the acceptor, has rendered it possible to perform a highly systematic mapping of the individual subsites of the ... | 1992 | 1627543 |
| crystal structure of the alkaline proteinase savinase from bacillus lentus at 1.4 a resolution. | savinase (ec3.4.21.14) is secreted by the alkalophilic bacterium bacillus lentus and is a representative of that subgroup of subtilisin enzymes with maximum stability in the ph range 7 to 10 and high activity in the range 8 to 12. it is therefore of major industrial importance for use in detergents. the crystal structure of the native form of savinase has been refined using x-ray diffraction data to 1.4 a resolution. the starting model was that of subtilisin carlsberg. a comparison to the struct ... | 1992 | 1738156 |
| a highly active and oxidation-resistant subtilisin-like enzyme produced by a combination of site-directed mutagenesis and chemical modification. | the subtilisins are known to be susceptible to chemical oxidation due to the conversion of met222 into the corresponding sulfoxide. a number of derivatives with resistance towards oxidation have previously been prepared by replacement of this group with the other 19 amino acid residues. unfortunately, the activities of these enzymes were of the order of 1-10% of that obtained with the wild-type enzyme. in contrast, the oxidation-labile cysteine mutant exhibited much higher activity, suggesting t ... | 1990 | 2269308 |
| crystallization and preliminary x-ray diffraction studies of an alkaline protease from bacillus lentus. | | 1988 | 3225854 |
| reduction of the 20-carbonyl group of c-21 steroids by spores of fusarium solani and other microorganisms. i. side-chain degradation, epoxide cleavage, and substrate specificity. | the spores of fusarium solani reduced the c(2)-carbonyl group, 1-dehydrogenated ring "a" and cleaved the side chain of 16alpha, 17alpha-oxidopregn-4-ene-3, 20-dione (16alpha, 17alpha-oxidoprogesterone)(i) to give the following products: 20alpha-hydroxy-16alpha, 17alpha-oxidopregn-4-en-3-one(ii); 20alpha-hydroxy-16alpha, 17alpha-oxidopregna-1, 4-dien-3-one(iii); 16alpha-hydroxy-17a-oxa-androsta-1, 4-diene-3, 17-dione (16alpha-hydroxy-1-dehydrotestololactone)(iv); and 16alpha, 17beta-dihydroxy-and ... | 1972 | 5021973 |
| [base composition of dna and phenotypic characterization of bacillus firmus and bacillus lentus]. | | 1982 | 7187508 |
| 1h, 13c and 15n nmr backbone assignments and secondary structure of the 269-residue protease subtilisin 309 from bacillus lentus. | 1h, 13c and 15n nmr assignments of the backbone atoms of subtilisin 309, secreted by bacillus lentus, have been made using heteronuclear 3d nmr techniques. with 269 amino acids, this protein is one of the largest proteins to be sequentially assigned by nmr methods to date. because of the size of the protein, some useful 3d correlation experiments were too insensitive to be used in the procedure. the hnco, hn(co)-ca, hnca and hcaco experiments are robust enough to provide most of the expected cor ... | 1994 | 8019137 |
| cavity mutants of savinase. crystal structures and differential scanning calorimetry experiments give hints of the function of the buried water molecules in subtilisins. | the subtilisin molecule possesses several internal water molecules, which may be characterised as an integral part of the protein structure. we have introduced specific mutations (t71i, t71s, t71v, t71a and t71g) at position 71 in the subtilisin variant savinase from bacillus lentus. this position is involved in a hydrogen bonded network with several internal water molecules, forming a water channel. the water channel and most of the other internal water molecules are positioned in the interface ... | 1994 | 8089841 |
| mutational replacements of the amino acid residues forming the hydrophobic s4 binding pocket of subtilisin 309 from bacillus lentus. | the amino acid side chains of ile107, leu126, and leu135 participate in the formation of the important hydrophobic s4 binding pocket of the subtilisin savinase. ile107 and leu126, located on each side of the pocket, point toward each other, and leu135 is situated at the bottom of the pocket. these amino acid residues have been substituted for other hydrophobic amino acid residues by site-directed mutagenesis, and the resulting enzymes have been characterized with respect to their p4 substrate pr ... | 1993 | 8369272 |
| significance of hydrophobic s4-p4 interactions in subtilisin 309 from bacillus lentus. | the subtilisins have an extended substrate binding cleft comprising at least 8 subsites. two pockets at the s1 and s4 sites are particularly conspicuous, and the interactions between substrate and these two pockets are very important for the substrate specificity. phe residues have mutationally been introduced at one of positions 102, 128, 130, and 132 of the subtilisin savinase from bacillus lentus to investigate the effects of introducing bulky groups along the rim of the s4 binding pocket. it ... | 1993 | 8457550 |
| backbone dynamics of the 269-residue protease savinase determined from 15n-nmr relaxation measurements. | backbone dynamics of savinase, a subtilisin of 269 residues secreted by bacillus lentus, have been studied using 15n relaxation measurements derived from proton-detected dimensional 1h-15n-nmr spectroscopy. 15n spin-lattice rate constants (r1), spin-spin relaxation-rate constants(r2), and 1h-15n nuclear overhauser effects (noe) were determined for 84% of the backbone amide 15n nuclei. the model-free formalism [lipari, g. & szabo, a. (1982) j. am. chem. soc. 104, 4546-4559] was used to derive val ... | 1996 | 8654411 |
| crystal structures of the alkaline proteases savinase and esperase from bacillus lentus. | savinase and esperase (ec. 3.4.21.14) are secreted by the alkalophilic bacterium bacillus lentus and are representatives of that subgroup of subtilisin enzymes with maximum stability in the range of ph 7 to 10 and high activity in the range of ph 8 to 12. the crystal structures of native savinase and diisopropyl fluorophosphate (dfp) inhibited esperase have been refined using x-ray data to 1.4 angstroms and 1.8 angstroms resolution respectively collected with synchrotron radiation. the structure ... | 1996 | 8796310 |
| studies of binding sites in the subtilisin from bacillus lentus by means of site directed mutagenesis and kinetic investigations. | | 1996 | 8796314 |
| substrate specificity of natural variants and genetically engineered intermediates of bacillus lentus alkaline proteases. | three natural variants of subtilisin lentus could be differentiated by their amino acid sequence and their specific activity with low molecular weight peptide substrates of the type saapfpna. the variants had amino acid exchanges in five, respective six positions of their amino acid sequence, four of which are located in the substrate loop of the enzyme (positions 92 - 102). variants of one type of highly alkaline subtilisin (subtilisin 309) were made by site directed mutagenesis, each containin ... | 1996 | 8796329 |
| unusual ligand binding at the active site domain of an engineered mutant of subtilisin bl. | as an attempt to recruit the third calcium binding site of thermitase into subtilisin bl, a bacillus lentus alkaline protease (blap), the amino acid sequence from position 50 to 60 and position 92 was modified to the equivalent amino acids in thermitase. the resulting protein, designated blapm109, exhibited unusual biochemical features. peptide mapping and gel electrophoresis revealed that two protein species co-purify in a ratio of about 1:1. form 1 consisted of a single polypeptide of 269 amin ... | 1996 | 8796330 |
| structural changes leading to increased enzymatic activity in an engineered variant of bacillus lentus subtilisin. | | 1996 | 8796332 |
| active-site titration of serine proteases using a fluoride ion selective electrode and sulfonyl fluoride inhibitors. | we report a general procedure for the determination of active enzyme concentrations for serine proteases. the method relies on the measurement of fluoride ion released from sulfonyl fluorides upon reaction with the active-site serine using an ion selective electrode. the results have been independently confirmed by amino acid analyses of subtilisins and by spectrofluorometric and spectrophotometric titrations. the minimal enzyme concentration detectable is 1-10 microm protease. the method is ins ... | 1996 | 8937565 |
| a low-barrier hydrogen bond in subtilisin: 1h and 15n nmr studies with peptidyl trifluoromethyl ketones. | the n delta 1 proton of his 64 forms a hydrogen bond with asp 32, as part of the catalytic triad in serine proteases of the subtilisin family. his 64 in subtilisin has been studied by 1h and 15n nmr spectroscopy in the presence and absence of peptidyl trifluoromethyl ketones (tfmks) that are transition state analog inhibitors. for subtilisin carlsberg, the downfield resonance of the imidazolium n delta 1 proton is approximately 18.3 ppm and the d/h fractionation factor is 0.55 +/- 0.04 at ph 5.5 ... | 1996 | 8961961 |
| site-directed mutagenesis combined with chemical modification as a strategy for altering the specificity of the s1 and s1' pockets of subtilisin bacillus lentus. | by combining site-directed mutagenesis with chemical modification, we have altered the s1 and s1' pocket specificity of subtilisin bacillus lentus (sbl) through the incorporation of unnatural amino acid moieties, in the following manner: wt --> cysmutant + h3cso2sr --> cys-sr, where r may be infinitely variable. a paradigm between extent of activity changes and surface exposure of the modified residue has emerged. modification of m222c, a buried residue in the s1' pocket of sbl, caused dramatic ... | 1998 | 9558332 |
| bacillus pseudomycoides sp. nov. | previous dna relatedness studies showed that strains identified as bacillus mycoides segregated into two genetically distinct yet phenotypically similar groups, one being b. mycoides sensu stricto and the other, an unclassified taxon. in the present study, the taxonomic position of this second group was assessed by measuring dna relatedness and determining phenotypic characteristics of an increased number of b. mycoides strains. also determined was the second group's 16s rna gene sequence. the 3 ... | 1998 | 9734060 |
| the 0.78 a structure of a serine protease: bacillus lentus subtilisin. | ultrahigh-resolution x-ray diffraction data from cryo-cooled, b. lentus subtilisin crystals has been collected to a resolution of 0.78 a. the refined model coordinates have a rms deviation of 0.22 a relative to the same structure determined at room temperature and 2.0 a resolution. several regions of main-chain and side-chain disorder have been identified for 21 out of 269 residues in one polypeptide chain. hydrogen atoms appear as significant peaks in the fo - fc difference electron density map ... | 1998 | 9753430 |
| a combinatorial approach to chemical modification of subtilisin bacillus lentus. | the reaction between methanethiosulfonate reagents and cysteine mutants of subtilisin is quantitative and can be used to prepare chemically modified mutant enzymes (cmms) with novel properties. the virtually unrestricted structural variations possible for cmms presents a preparative and screening challenge. to address this, a rapid combinatorial method for preparing and screening the activities of cmms has been developed. | 1998 | 9873530 |
| engineered bacillus lentus subtilisins having altered flexibility. | the three-dimensional structures of engineered variants of bacillus lentus subtilisin having increased enzymatic activity, k27r/n87s/v104y/n123s/t274a (rsysa) and n76d/n87s/s103a/v104i (dsai), were determined by x-ray crystallography. in addition to identifying changes in atomic position we report a method that identifies protein segments having altered flexibility. the method utilizes a statistical analysis of variance to delineate main-chain temperature factors that represent significant depar ... | 1999 | 10493860 |
| altered flexibility in the substrate-binding site of related native and engineered high-alkaline bacillus subtilisins. | high-alkaline serine proteases have been successfully applied as protein degrading components of detergent formulations and are subject to extensive protein engineering efforts to improve their stability and performance. dynamics has been suggested to play an important role in determining enzyme activity and specificity and it is therefore of interest to establish how local changes in internal mobility affect protein stability, specificity and performance. here we present the dynamic properties ... | 1999 | 10493861 |
| contribution of the cell wall component teichuronopeptide to ph homeostasis and alkaliphily in the alkaliphile bacillus lentus c-125. | a teichuronopeptide (tup) is one of major structural components of the cell wall of the facultative alkaliphilic strain bacillus lentus c-125. a mutant defective in tup synthesis grows slowly at alkaline ph. an upper limit of ph for growth of the mutant was 10.4, while that of the parental strain c-125 was 10.8. gene tupa, directing synthesis of tup, was cloned from c-125 chromosomal dna. the primary translation product of this gene is likely a cytoplasmic protein (57. 3 kda) consisting of 489 a ... | 1999 | 10542159 |
| the controlled introduction of multiple negative charge at single amino acid sites in subtilisin bacillus lentus. | the use of methanethiosulfonates as thiol-specific modifying reagents in the strategy of combined site-directed mutagenesis and chemical modification allows virtually unlimited opportunities for creating new protein surface environments. as a consequence of our interest in electrostatic manipulation as a means of tailoring enzyme activity and specificity, we have adopted this approach for the controlled incorporation of multiple negative charges at single sites in the representative serine prote ... | 1999 | 10632039 |
| altering the specificity of subtilisin bacillus lentus through the introduction of positive charge at single amino acid sites. | the use of methanethiosulfonates as thiol-specific modifying reagents in the strategy of combined site-directed mutagenesis and chemical modification allows virtually unlimited opportunities for creating new protein surface environments. as a consequence of our interest in electrostatic manipulation as a means of tailoring enzyme activity and specificity, we have recently adopted this approach for the controlled incorporation of multiple negative charges at single sites in the representative ser ... | 1999 | 10632040 |
| cloning and sequencing of an alkaline protease gene from bacillus lentus and amplification of the gene on the b. lentus chromosome by an improved technique. | a gene encoding an alkaline protease was cloned from an alkalophilic bacillus, and its nucleotide sequence was determined. the cloned gene was used to increase the copy number of the protease gene on the chromosome by an improved gene amplification technique. | 2000 | 10653758 |
| display of active subtilisin 309 on phage: analysis of parameters influencing the selection of subtilisin variants with changed substrate specificity from libraries using phosphonylating inhibitors. | many attempts have been made to endow enzymes with new catalytic activities. one general strategy involves the creation of random combinatorial libraries of mutants associated with an efficient screening or selection scheme. phage display has been shown to greatly facilitate the selection of polypeptides with desired properties by establishing a close link between the polypeptide and the gene that encodes it. selection of phage displayed enzymes for new catalytic activities remains a challenge. ... | 2000 | 10656819 |
| benzophenone boronic acid photoaffinity labeling of subtilisin cmms to probe altered specificity. | a transition state analogue inhibitor, boronic acid benzophenone (bbp) photoprobe, was used to study the differences in the topology of the s1 pocket of chemically modified mutant enzymes (cmms). the bbp proved to be an effective competitive inhibitor and a revealing active site directed photoprobe of the cmms of the serine protease subtilisin bacillus lentus (sbl) which were chemically modified with the hydrophobic, negatively charged and positively charged moieties at the s1 pocket s166c resid ... | 2000 | 10732973 |
| controlled site-selective protein glycosylation for precise glycan structure-catalytic activity relationships. | glycoproteins occur naturally as complex mixtures of differently glycosylated forms which are difficult to separate. to explore their individual properties, there is a need for homogeneous sources of carbohydrate-protein conjugates and this has recently prompted us to develop a novel method for the site-selective glycosylation of proteins. the potential of the method was illustrated by site-selective glycosylations of subtilisin bacillus lentus (sbl) as a model protein. a representative library ... | 2000 | 10976501 |
| site-selective glycosylation of subtilisin bacillus lentus causes dramatic increases in esterase activity. | using site directed mutagenesis combined with chemical modification, we have developed a general and versatile method for the glycosylation of proteins which is virtually unlimited in the scope of proteins and glycans that may be conjugated and in which the site of glycosylation and the nature of the introduced glycan can be carefully controlled. we have demonstrated the applicability of this method through the synthesis of a library of 48 glycosylated forms of the serine protease subtilisin bac ... | 2000 | 10976502 |
| covalent modification of subtilisin bacillus lentus cysteine mutants with enantiomerically pure chiral auxiliaries causes remarkable changes in activity. | methanethiosulfonate reagents may be used to introduce virtually unlimited structural modifications in enzymes via reaction with the thiol group of cysteine. the covalent coupling of enantiomerically pure (r) and (s) chiral auxiliary methanethiosulfonate ligands to cysteine mutants of subtilisin bacillus lentus induces spectacular changes in catalytic activity between diastereomeric enzymes. amidase and esterase kinetic assays using a low substrate approximation were used to establish kcat/km va ... | 2000 | 11003141 |
| differences in the specificities of the highly alkalophilic proteinases savinase and esperase imposed by changes in the rigidity and geometry of the substrate binding sites. | savinase and esperase are closely related highly alkalophilic proteinases produced by bacillus lentus. they are suitable couple for investigating the structural basis of proteinase specificity due to the identity of the catalytic and the differences in the substrate binding sites. two of the substitutions in these sites are very important: t129p and g131p. the two prolines provide an extra rigidity of the savinase-binding site. the substitutions s166n and q191t in the s1 recognition loop change ... | 2001 | 11370841 |
| substrate specificity of the highly alkalophilic bacterial proteinase esperase: relation to the x-ray structure. | esperase is a highly alkalophilic bacterial proteinase produced by bacillus lentus. the enzyme hydrolyzes peptide bonds comprising the carboxylic groups of hydrophobic as well as hydrophilic residues in the oxidized insulin b chain. some of these bonds are not attacked by other alkaline microbial proteinases. p1-p4 specificity was determined by a series of peptide nitroanilides. the s1 recognition loop exhibits a preference for phe. the "cleft" of the smallest subsite s2 prefers ala and exhibits ... | 2001 | 11400059 |
| chemical composition and cytotoxic and antimicrobial activity of calycotome villosa (poiret) link leaves. | the chemical composition of the essential oil and methanol extract of calycotome villosa (poiret) link leaves collected in sardinia (italy) has been studied by analytical and spectroscopic methods. falcarinol and some alcohols, terpenes, furan derivatives, and paraffins have been isolated from the essential oil. thirteen alkaloids and falcarinol have been identified in the chloroform fraction of the basic methanol extract. six flavonoids and four anthraquinones have been isolated in the chlorofo ... | 2001 | 11482772 |
| antimicrobial evaluation of coumarins and flavonoids from the stems of daphne gnidium l. | the antimicrobial activity of stems methanol extract from daphne gnidium l. collected from sardinia (italy) was evaluated against 6 strains of standard and clinical isolated gram (+/-) bacteria. the antimicrobial effect on two strains of fungi was also tested. the extract in toto exhibited antibacterial activity against bacillus lentus and escherichia coli, but was inactive against fungi. four coumarins (daphnetin, daphnin, acetylumbelliferone, daphnoretin) and seven flavonoids (luteolin, orient ... | 2001 | 11515721 |
| microbial community analysis of thermophilic contact oxidation process by using ribosomal rna approaches and the quinone profile method. | microbial community structure of a lab scale thermophilic aerobic wastewater treatment reactor was analyzed by a combination of culture-independent methods. quinone profile method provides for chemical analysis of respiratory quinone molecular species, which corresponds to bacterial groups. denaturing gradient gel electrophoresis (dgge) of pcr-amplified 16s rdna partial sequences (pcr-dgge) clarifies community changes at species level, as dgge can separate dna fragments of different sequences. c ... | 2002 | 11827349 |
| decrease in cytoplasmic ph-homeostastatic activity of the alkaliphile bacillus lentus c-125 by a cell wall defect. | cytoplasmic ph homeostatic activities of cell wall-defective derivatives of the alkaliphile bacillus lentus c-125 were assessed using a ph-sensitive fluorescent probe, bcecf. it was shown that the acidic cell wall components took part in maintenance of the cytoplasmic ph neutrality at alkaline ph. | 2002 | 11866113 |
| chemically modified "polar patch" mutants of subtilisin in peptide synthesis with remarkably broad substrate acceptance: designing combinatorial biocatalysts. | a significant enhancement of the applicability of the serine protease subtilisin bacillus lentus (sbl) in peptide synthesis was achieved by using the strategy of combined site-directed mutagenesis and chemical modification to create chemically modified mutant (cmm) enzymes. the introduction of polar and/or homochiral auxiliary substituents, such as x=oxazolidinones, alkylammonium groups, and carbohydrates at position 166 at the base of the primary specificity s(1) pocket created sbl cmms s166c-s ... | 2002 | 12298003 |
| do enzymes change the nature of transition states? mapping the transition state for general acid-base catalysis of a serine protease. | the properties of the transition state for serine protease-catalyzed hydrolysis of an amide bond were determined for a series of subtilisin variants from bacillus lentus. there is no significant change in the structure of the enzyme upon introduction of charged mutations s156e/s166d, suggesting that changes in catalytic activity reflect global properties of the enzyme. the effect of charged mutations on the pk(a) of the active site histidine-64 n(epsilon)(2)-h was correlated with changes in the ... | 2003 | 12962477 |
| bacillus galactosidilyticus sp. nov., an alkali-tolerant beta-galactosidase producer. | a novel bacillus isolate from raw milk and four strains from diverse origins that were identified previously as bacillus lentus, bacillus firmus and bacillus circulans showed a high degree of similarity in amplified rdna restriction analysis, sds-page and routine phenotypic tests, whilst 16s rdna sequence comparisons and dna relatedness data showed that this taxon was different from related bacillus species. on the basis of these data, bacillus galactosidilyticus sp. nov. is proposed, with the t ... | 2004 | 15023985 |
| engineering a substrate-specific cold-adapted subtilisin. | one region predicted to be highly flexible for a psychrophilic enzyme, ta39 subtilisin (s39), was transferred in silico to the mesophilic subtilisin, savinase (ec 3.4.21.62), from bacillus lentus (clausii). the engineered hybrid and savinase were initially investigated by molecular dynamic simulations at 300 k to show binding region and global flexibility. the predicted s39 region consists of 12 residues, which due to homology between the subtilisins, results in a total change of eight residues. ... | 2004 | 15047911 |
| bacillus farraginis sp. nov., bacillus fortis sp. nov. and bacillus fordii sp. nov., isolated at dairy farms. | forty-eight bacterial strains were isolated at dairy farms from raw milk, the milking apparatus, green fodder or feed concentrate after a heat treatment of 30 min at 100 degrees c. in this way, spore-forming bacteria with a very high intrinsic heat resistance were selected for. the aerobic spore-forming isolates were subjected to a polyphasic taxonomical study, including repetitive element sequence-based pcr typing, whole-cell protein profiling, 16s rdna sequence analysis, dna-dna hybridizations ... | 2004 | 15280314 |
| packing selection of bacillus lentus subtilisin and a site-specific variant. | the crystallization of a variant of bacillus lentus subtilisin and the native enzyme was achieved using identical conditions. the variant b. lentus was found to grow in two crystal forms, form 1 and form 2, whereas the native b. lentus subtilisin enzyme crystallized in only one, form 1. form 2 crystals, once obtained, were found to grow much more rapidly than form 1 crystals. the lattice contacts and structural changes giving both crystal forms have been examined. the results show that crystal f ... | 1994 | 15299431 |
| characterisation of the microflora of attiéké, a fermented cassava product, during traditional small-scale preparation. | attiéké is a fermented cassava product consumed mainly in cote d'ivoire. the aim of this study was to characterise the attiéké fermentation by examining products from 15 small-scale production sites at various stages of its preparation. for the preparation of attiéké, fresh cassava is grated to a pulp and inoculated with 10% of a spontaneous traditional inoculum. the inocula contained aerobic mesophiles at mean numbers of 8.2 x 10(7) cfu/g and lactic and acetic acids at mean concentrations of 0. ... | 2006 | 16213052 |
| pectinase activity of anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (diascorea rotundata). | anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (diascorea rotundata) were isolated by the looping-out method and found to consist of clostridium (three isolates), corynebacterium (three isolates), vibrio (one isolate), and bacillus lentus (one isolate). enzyme assay for hydrolase, lyase, and pectinesterase activities by the cup-plate method showed that except for vibrio sp., b. lentus, and two isolates of corynebacterium no pectinase activity could be detected for ... | 1981 | 16345726 |
| bio-deposition of a calcium carbonate layer on degraded limestone by bacillus species. | to obtain a restoring and protective calcite layer on degraded limestone, five different strains of the bacillus sphaericus group and one strain of bacillus lentus were tested for their ureolytic driven calcium carbonate precipitation. although all the bacillus strains were capable of depositing calcium carbonate, differences occurred in the amount of precipitated calcium carbonate on agar plate colonies. seven parameters involved in the process were examined: calcite deposition on limestone cub ... | 2006 | 16491305 |
| isolation, purification and properties of new restriction endonucleases from bacillus badius and bacillus lentus. | we tentatively named two enzymes as bbai and blei, which were isolated and purified from gram-positive mesophilic bacteria bacillus badius 1458 and bacillus lentus 1689 respectively, by ammonium sulphate precipitation, phosphocellulose and heparin-sepharose column chromatography. sds-page protein profiles for bbai and blei showed denatured molecular weights of 52 and 48 kda, respectively. bbai hydrolyzed puc18 dna into 1900 and 700 bp, pbr322 dna into two fragments of 2800 and 1500 bp and phix17 ... | 2007 | 16644193 |
| characterization of a new bacteriocin produced from a novel isolated strain of bacillus lentus ng121. | the new bacteriocin is produced from bacillus lentus ng121 isolated from khameera - a traditional fermented food from himachal pradesh, india which has been reported for the first time in the literature to produce bacteriocin and exhibited very high activity units of 20 x 10(5) au (arbitrary units)/ml. this bacteriocin was partially purified and was further characterized to assess its preservation characteristics. it showed strong antimicrobial activity against the most challenging and serious t ... | 2006 | 16779629 |
| in vitro inhibitory activity of probiotic spore-forming bacilli against genotoxins. | to investigate the ability of bacilli of various species (bacillus clausii, bacillus subtilis, bacillus lentus, bacillus pumilus. bacillus megaterium, bacillus firmus, bacillus sp.) and origins (probiotic and collection strains) to counteract the activity of some representative dna-reactive agents. | 2008 | 18194161 |
| structural and mutational analyses of the interaction between the barley alpha-amylase/subtilisin inhibitor and the subtilisin savinase reveal a novel mode of inhibition. | subtilisins represent a large class of microbial serine proteases. to date, there are three-dimensional structures of proteinaceous inhibitors from three families in complex with subtilisins in the protein data bank. all interact with subtilisin via an exposed loop covering six interacting residues. here we present the crystal structure of the complex between the bacillus lentus subtilisin savinase and the barley alpha-amylase/subtilisin inhibitor (basi). this is the first reported structure of ... | 2008 | 18556023 |
| decontamination of prion protein (bse301v) using a genetically engineered protease. | a previous study has demonstrated the potential of alkaline proteases to inactivate bovine spongiform encephalopathy (bse301v). here we explored the use of mc3, a genetically engineered variant of bacillus lentus subtilisin. mc3 was used to digest bse301v infectious mouse brain homogenate (imbh). mc3 eliminated all detectable 6h4-immunoreactive material at ph 10 and 12; however, proteinase k was only partially effective at ph 12. when bioassayed in vm mice, mc3- and proteinase k-digested imbh ga ... | 2009 | 19201054 |
| in vitro antimicrobial effect of satureja wiedemanniana against bacillus species isolated from raw meat samples. | in this study a total of 30 raw meat samples obtained from ankara, turkey were screened for the presence of bacillus species. among the meat samples analyzed, the predominant species isolated was bacillus circulans; other bacillus species were identified as bacillus firmus, bacillus lentus, bacillus megaterium, bacillus licheniformis, bacillus mycoides, bacillus sphaericus, and bacillus cereus. minced meat samples were more contaminated with bacillus species than sliced beef sample. from these s ... | 2009 | 19735196 |
| evaluation of ultrasound velocity measurements for estimating protease activities using casein as substrate. | ultrasonic resonator technology (urt) was compared with the well established uv-vis/ninhydrin assay to estimate protease activities in defined buffer systems. hydrolysis of casein was measured using subtilisin, trypsin, halophilic protease from haloferax mediterranei and bacillus lentus alkaline protease. sensitivity, reproducibility, working range as well as the limit of detection and the limit of quantification were comparable for both methods. salt concentrations (0.5 m nacl) interfered with ... | 2010 | 19821076 |
| the effect of a beta-mannanase-based enzyme on growth performance and humoral immune response of broiler chickens fed diets containing graded levels of whole dates. | the use of low-quality whole dates in broilers' diets was evaluated. the apparent metabolizable energy (amen) of whole dates and date pits was determined with or without a beta-mannanase-based enzyme (hemicell--a registered trademark of chemgen corp., gaithersburg, md. dried bacillus lentus fermentation solubles with 158 million units/kg minimum enzyme activity. recommended usage rate is 0.4 g kg(-1) of feed. chemgen corp., gaithersburg, md, usa). a 3 x 2 factorial arrangement consisting of whol ... | 2010 | 20383608 |
| microbiological studies of ethnic meat products of the eastern himalayas. | native microorganisms from some ethnic meat products of the eastern himalayas such as lang kargyong, yak kargyong, faak kargyong, lang satchu, yak satchu and suka ko masu were isolated and characterized. the bacterial isolates included lactobacillus sake, lactobacillus curvatus, lactobacillus divergens, lactobacillus carnis, lactobacillus sanfrancisco, lactobacillus plantarum, lactobacillus casei, lactobacillus brevis, enterococcus faecium, leuconostoc mesenteroides, pediococcus pentosaceous, ba ... | 2010 | 20416835 |
| purification and characterization of a surfactant-stable high-alkaline protease from bacillus sp. b001. | the newly isolated alkalophilic bacillus sp. b001 produced a high level of proteolytic activity (34277 u/ml) when grown in production medium, and a 28 kda protease, designated aprb, was purified from the culture supernatant. partial amino acid sequences were obtained by tandem mass spectrometry (ms/ms) and a pair of degenerate primers was developed to amplify a 467-bp genomic sequence. the observed and predicted amino acid sequences showed similarity with sequences of high-alkaline proteases fro ... | 2010 | 20417096 |
| bacillus graminis sp. nov., an endophytic bacterium isolated from a coastal dune plant. | a gram-positive endophytic bacterium, designated strain yc6957t, was isolated from surface-sterilized root of a halophyte (elymus mollis trin.), inhabiting a coastal tidal flats of namhae island, located at southern end in korea and was subjected to a polyphasic taxonomic study. cells were facultatively anaerobic, endospore-forming, rods to coccoid rods in shape and motile by a single flagellum. strain yc6957t was catalase-positive, oxidase-negative and able to grow at nacl concentrations in the ... | 2010 | 20656804 |
| mechanistic investigation of decolorization and degradation of reactive red 120 by bacillus lentus bi377. | bacillus lentus bi377, isolated from textile effluent-contaminated soil, was able to degrade 97% and 92% of reactive red 120 dye when 1200 and 1500 mg/l, respectively, of dye was added to nutrient broth (nb) at 35 °c within 12 h. uv-vis spectroscopy, gc-ms, ftir and 1h nmr revealed the formation of catechol which may be further utilized by the bacterium via the tca cycle, leading to complete mineralization. structural analysis of metabolites in conjunction with enzyme activity studies confirmed ... | 2010 | 20864334 |
| antibacterial activity of the essential oil of satureja wiedemanniana against bacillus species isolated from chicken meat. | the antimicrobial activity of the essential oil (eo) of satureja wiedemanniana against bacillus spp. isolated from chicken meat was investigated. thirty-seven bacillus strains were isolated from 15 chicken meat samples and examined for proteolytic and lipolytic activities by agar well diffusion assay. of 37 bacillus isolated from raw chicken samples, which based on a clear zone diameter of ≥ 6 mm in agar well diffusion assays for proteolytic activity, 19 bacillus strains were selected for this s ... | 2010 | 20932089 |
| design of a recombinant escherichia coli for producing l-phenylalanine from glycerol. | a recombinant escherichia coli was engineered to produce the commercially important amino acid l-phenylalanine (l-phe) using glycerol as the carbon source. compared to the conventionally used glucose and sucrose, glycerol is a less expensive carbon source. as phenylalanine dehydrogenase (phedh) activity is involved in the last step of l-phe synthesis in e. coli, a phenylalanine dehydrogenase gene (phedh) from the thermotolerant bacillus lentus was cloned into prsfduet-1 (pphedh) and expressed in ... | 2012 | 22806734 |
| bacillus panacisoli sp. nov., isolated from ginseng soil. | a gram-staining-positive, motile, facultatively anaerobic, endospore-forming and rod-shaped bacterium, designated strain cj32(t), was isolated from ginseng soil at geumsan in korea. the isolate grew optimally at 30 °c, 2% (w/v) nacl and ph 7.0. colonies of strain cj32(t) were beige and circular with an entire margin on lb agar plates. phylogenetic analysis based on 16s rrna gene sequences indicated that strain cj32(t) was associated with the genus bacillus and was most closely related to bacillu ... | 2014 | 24277860 |
| purification and characterization of cell wall-associated n-acetylmuramyl-l-alanine amidase from alkaliphilic bacillus lentus c-125. | cells of the facultative alkaliphile bacillus sp. c-125 grown at neutral ph autolyze rapidly in alkaline buffers of ph 9-10. alkaline autolytic activity has been found mainly in the cell wall fraction. a peptidoglycanlytic enzyme was extracted from the cell wall fraction suspended in 4m licl. the enzyme was identified as n-acetylmuramyl-l-alanine amidase, with a molecular mass of 58kda. at low salinity, the enzyme formed an aggregate of high molecular mass. the peptidoglycan lytic reaction of th ... | 1996 | 27299715 |
| engineering of isoamylase: improvement of protein stability and catalytic efficiency through semi-rational design. | isoamylase catalyzes the hydrolysis of α-1,6-glycosidic linkages in glycogen, amylopectin and α/β-limit dextrins. a semi-rational design strategy was performed to improve catalytic properties of isoamylase from bacillus lentus. three residues in vicinity of the essential residues, arg505, asn513, and gly608, were chosen as the mutation sites and were substituted by ala, pro, glu, and lys, respectively. thermal stability of the mutant r505p and acidic stability of the mutant r505e were enhanced. ... | 2016 | 26597030 |
| analysis of an industrial production suspension of bacillus lentus subtilisin crystals by powder diffraction: a powerful quality-control tool. | a microcrystalline suspension of bacillus lentus subtilisin (savinase) produced during industrial large-scale production was analysed by x-ray powder diffraction (xrpd) and x-ray single-crystal diffraction (mx). xrpd established that the bulk microcrystal sample representative of the entire production suspension corresponded to space group p212121, with unit-cell parameters a = 47.65, b = 62.43, c = 75.74 å, equivalent to those for a known orthorhombic crystal form (pdb entry 1ndq). mx using syn ... | 2014 | 24699655 |
| enzyme based cleavage strategy of bacillus lentus bi377 in response to metabolism of azoic recalcitrant. | bacillus lentus bi377 (b. lentus bi377) an alkaliphilic strain has accomplished the discriminate color removal strategy for reactive red sulfonated azoic recalcitrant irrespective of their molecular structure. during the decolorization experiment, it was observed that the diazo dye first followed chromophoric cleavage by azoreductase via typical azoreduction whereas, in case of monoazo dye, cleavage took place by peroxidase via successive electron transfers to oxide surface resulting in the asym ... | 2013 | 23313681 |