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cellulase location in cellvibrio fulvus.the location of cellulase in c. fulvus depends on the carbon source for growth and the age of the culture. when cells were grown on glucose or cellobiose all cmc-hydrolyzing enzyme was cell-bound but only part of the activity was located on the cell surface. treatment of cells with edta, lysozyme, and detergents and subsequent fractionation experiments showed that cellulase was also located in the periplasm and bound to a membrane fraction. growth on cellulose gave cell-free cellulase active aga ...1975163668
the ultrastructure of the cellulolytic bacterium cellvibrio fulvus.thin sections of the cellulolytic bacterium cellvibrio fulvus were investigated before and after treatment with polymyxin b and triton x-100. the gram-negative vibrio appeared to have no separate stained g2 layer between the l and c membranes. cells lysed after treatment with polymyxin or swelled in the presence of 5% sucrose. blebs were formed from the l membrane, and the inner structure of the cells changed. triton x-100 lysed cells even in the presence of 20% sucrose. the flagellum disappeare ...1975163671
regulation of cellvibriocin activity.attempts were made to enhance the production of cellvibriocin, the bacteriocin produced by cellvibrio sp. ncib 9916, by a study of cultural conditions in agar (solid) media. in such media cellvibriocin was unstable: within 24 h all activity disappeared at 45 degrees c, 70% at 37 degrees c and more than 50% within 5 days at 5 degrees c. the temperature at which the producer strain was grown was critical, 25 degrees c being optimal. cellvibriocin activity was markedly affected by the composition o ...1977617829
extracellular endo-beta-1,4-glucanase in cellvibrio vulgaris.endo-beta-1,4-glucanase of the cellulolytic bacterium cellvibrio vulgaris is an actively secreted, truly extracellular enzyme, as supported by growth and secretion studies using filter paper as the sole carbon source.1978697358
cellulase induction and the use of cellulose as a preferred growth substrate by cellvibrio gilvus.cellvibrio gilvus produced cellulase when grown in the presence of cellulose or carboxymethyl cellulose (cmc) but not when grown in the presence of glucose or cellobiose. this was so whether or not these compounds were the sole carbon sources present. repeated addition of small amounts of glucose prevented cellulase formation in the presence of cellulose. it is concluded that cellulose and cmc induce cellulase formation and glucose and cellobiose repressit. cellulose stimulated growth when added ...19761009505
optimum culture conditions for the epoxidation of cis-propenylphosphonate to fosfomycin by cellvibrio gilvus.approximately 470 strains of various microorganisms were tested for their ability to epoxidize cis-propenylphosphonate (ppoh) to (-)-cis-1,2-epoxypropylphosphonate (fosfomycin, fom). cellvibrio gilvus ky 3412 was selected as the best strain. to obtain higher activity, fom-resistant strains were derived by n-methyl-n'-nitro-n-nitrosoguanidine mutagenesis. mutant ky 3413, showing ten times higher fom resistance, was selected. the conditions for the conversion of ppoh to fom during the cultivation ...19921368198
purification and properties of a xylanase from cellvibrio gilvus that hydrolyzes p-nitrophenyl cellooligosaccharides.an enzyme component that hydrolyzes pnp-g2 but not cmc has been isolated from a culture broth of cellvibrio gilvus by a multi-step procedure involving butyl-toyopearl, deae-toyopearl, and cm-toyopearl chromatographies. the purified enzyme gave a single protein band on native, sds-, and ief-page. the enzyme had a molecular weight of 40,000, an isoelectric point of 5.0, an optimum ph of 6.5, and an optimum temperature of 55 degrees c. it was stable from ph 4.0 to 9.0 at 37 degrees c for 1 hr and b ...19911368727
characterization of a beta-glucosidase encoded by a gene from cellvibrio gilvus. 19911368758
synthetic reaction of cellvibrio gilvus cellobiose phosphorylase.the synthetic reactions of the cellobiose phosphorylase from cellvibrio gilvus were investigated in detail. it was found that, besides d-glucose, some sugars having substitution or deletion of the hydroxyl group at c2 or c6 of the d-glucose molecule could serve as a glucosyl acceptor, though less effectively than d-glucose. the enzyme showed higher activity with beta-d-glucose than with the alpha-anomer as an acceptor. this result indicates that it recognizes the anomeric hydroxyl group not invo ...19921429509
electronmicroscopie observations on the degradation of cellulose fibres by cellvibrio fulvus and sporocytophaga myxococcoides. 19724558950
antimicrobial effects of cellvibrio on blue-green algae. 19724626336
growth and cellulase formation by cellvibrio fulvus. 19725049544
dna sequence analysis of endoglucanase genes from pseudomonas fluorescens subsp. cellulosa and pseudomonas sp. ncib 8634.the dna of two previously isolated recombinant clones, one from pseudomonas sp. ncib 8634 (= cellvibrio mixtus) (ppc71) and another from pseudomonas fluorescens subsp. cellulosa (ppfc4) that express endoglucanase activity in e. coli was sequenced. plasmid ppc71 had three open reading frames, two of which include portions of plasmid pbr322. the third open reading frame occurs entirely within the pseudomonas dna insert and encodes a protein with a molecular mass of 5845 da. the dna insert in ppfc4 ...19901366996
sensitivity of cellulolytic bacteria to antibiotics.the sensitivity of eight cellulolytic bacterial strains to eight antibiotics was tested. the results showed that, in general, the strains belonging to cytophaga, cellvibrio, and cellfalcicula are more sensitive to antibiotics than those strains that belong to sporocytophaga and cellulomonas. the inhibitory activity of the tested antibiotics, though differing with different strains, showed the following categories: tetracycline, erythromycin, and chloromycetin were most active, kanamycin, strepto ...1977414477
characteristics of a subcellular system from cellvibrio gilvus for the incorporation of amino acids into protein. 19685644628
genes from cellvibrio mixtus encoding beta-1,3 endoglucanase.two genes encoding beta-1,3 glucanase activity were cloned from the gram-negative soil bacterium cellvibrio mixtus. the two clones, designated cwd (cell wall degradation) and lam (laminarin degradation), had distinct endonuclease restriction patterns and encoded enzymes with distinct substrate specificities. the 3.7-kilobase cwd insert encoded an enzyme which degraded yeast cell walls as well as the soluble beta-1,3 glucan laminarin and the insoluble beta-1,3 glucans zymosan and pachyman. the 1. ...19902285322
construction of chimeric beta-glucosidases with improved enzymatic properties.the amino acid sequences of beta-glucosidases from cellvibrio gilvus and agrobacterium tumefaciens show about 40% similarity. the ph/temperature optima and stabilities and substrate specificities of the two enzymes are quite different. c. gilvus beta-glucosidase exhibits an optimum ph of 6.2-6.4 and temperature of 35 degrees c, whereas the corresponding values for a. tumefaciens are 7.2-7.4 and 60 degrees c, respectively. the substrate specificity of a. tumefaciens enzyme toward different aryl g ...19957665615
novel cellulose-binding domains, nodb homologues and conserved modular architecture in xylanases from the aerobic soil bacteria pseudomonas fluorescens subsp. cellulosa and cellvibrio mixtus.to test the hypothesis that selective pressure has led to the retention of cellulose-binding domains (cbds) by hemicellulase enzymes from aerobic bacteria, four new xylanase (xyn) genes from two cellulolytic soil bacteria, pseudomonas fluorescens subsp. cellulosa and cellvibrio mixtus, have been isolated and sequenced. pseudomonas genes xyne and xynf encoded modular xylanases (xyle and xylf) with predicted m(r) values of 68,600 and 65000 respectively. xyle contained a glycosyl hydrolase family 1 ...19957492333
location of cellulase activity in cellvibrio gilvus. 19676076216
purification and properties of cellvibrio gilvus cellobiose phosphorylase.the cellobiose phosphorylase (ec 2.4.1.20) of cellvibrio gilvus, which is an endocellular enzyme, has been purified 196-fold with a recovery of 11% and a specific activity of 27.4 mumol of glucose 1-phosphate formed/min per mg of protein. the purification procedure includes fractionation with protamine sulphate, and hydroxyapatite and deae-sephadex a-50 chromatography. the enzyme appears homogeneous on polyacrylamide-gel electrophoresis, and a molecular weight of 280 000 was determined by molecu ...19836223623
synthesis of three hetero disaccharides, 4-o-beta-glucopyranosyl-6-deoxy-d-glucose, 4-o-beta-d-glucopyranosyl-d-mannosamine, and 4-o-beta-d-glucopyranosyl-d-mannose, and confirmation of their structures by c-13 nmr and ms.the three disaccharides, 4-o-beta-d-glucopyranosyl-6-deoxy-d-glucose (glc-6-deoxy-glc), 4-o-beta-d-glucopyranosyl-d-mannosamine (glc-mann), 4-o-beta-d-glucopyranosyl-d-mannose (glc-man), were synthesized from equimolar amounts of 6-deoxy-glucose and alpha-d-glucose-1-phosphate (g-1-p), d-mannosamine and alpha-d-glucose-1-phosphate (g-1-p), and d-mannose and alpha-d-glucose-1-phosphate (g-1-p), respectively, using cellobiose phosphorylase from cellvibrio gilvus. the yields were 60%, 10% and 50% b ...19957677766
construction and characterization of a chimeric beta-glucosidase.the amino acid sequences of beta-glucosidases from cellvibrio gilvus and agrobacterium tumefaciens show significant similarity in most of the parts. however, the ph/temperature optima and stabilities of the two enzymes are quite different. c. gilvus beta-glucosidase exhibits an optimum ph of 6.2-6.4 and temperature of 35 degrees c, whereas the corresponding values for a. tumefaciens are 7.2-7.4 and 60 degrees c respectively. to analyse these properties further, a chimeric beta-glucosidase was co ...19957848268
a gene encoding an exo-beta-glucosidase from cellvibrio mixtus.cellvibrio mixtus produces an array of endohydrolytic enzymes involved in the initial phases of beta-glycan polysaccharide degradation in the soil. these enzymes convert complex, high-molecular-weight, insoluble polysaccharides into low-molecular-weight, soluble oligosaccharides which must be further degraded for cellular uptake and catabolism. little is known about the enzymes involved in this latter process in c. mixtus. in this paper we report the cloning of the lam2 gene, which encodes an ex ...19979290063
a cellobiose phosphorylase from cellvibrio gilvus recognizes only the beta-d-form of 5a-carba-glucopyranose. 19938221729
characterization of an endo-1,3(4)-beta-d-glucanase gene from cellvibrio mixtus.an endo-1,3(4)-beta-d-glucanase gene (cwd2) of cellvibrio mixtus encoding laminarinase activity was cloned on a 3.9-kb psti fragment. the cwd2 enzyme, extracted from recombinant escherichia coli, degraded both beta-1,3 glucans and beta-1,3-1,4 mixed-linkage glucans, was endohydrolytic and so conformed to the enzyme class 3.2.1.6. the ph and temperature optima of the enzyme were approximately 7 and 40 degrees c respectively. the m(r) of specifically labelled cwd2 was approximately 34,000. this ge ...19938339916
cloning and characterization of two closely linked cellulase genes from cellvibrio mixtusa 16.5-kb bamhi fragment of the cellvibrio mixtus chromosome was found to direct carboxymethylcellulase, xylanase, and avicel hydrolysis. two closely linked genes were subcloned from this insert. the gene, cmci, was cloned as a 2.7-kb fragment and expressed in escherichia coli. it encoded an enzyme of approximately 74 kda which degraded carboxymethylcellulose and xylan but did not attack the microcrystalline cellulose substrate avicel. a second cellulase capable of degrading avicel, encoded by e ...19968661691
acceptor specificity of cellobiose phosphorylase from cellvibrio gilvus: synthesis of three branched trisaccharides.cellobiose phosphorylase from cellvibrio gilvus was examined for its acceptor specificity in the synthetic reaction with glucose-1-phosphate, using substrates in which the c-6 substituent of d-glc had been altered. a range of disaccharides were also tested for acceptor specificity but only those with (1-->6)-linkages were successful acceptors. melibiose, gentiobiose, isomaltose and also the monosaccharide glucuronamide were found to react with cellobiose phosphorylase and glucose-1-phosphate giv ...19989711833
possible roles for a non-modular, thermostable and proteinase-resistant cellulase from the mesophilic aerobic soil bacterium cellvibrio mixtus.the widespread presence of cellulose-binding domains in cellulases from aerobic bacteria and fungi suggests the existence of a strong selective pressure for the retention of these non-catalytic modules. the complete nucleotide sequence of the cellulase gene, cela, from the aerobic soil bacterium cellvibrio mixtus, was determined. it revealed an open reading frame of 1089 bp that encoded a polypeptide, defined as cellulase a (cela), of m(r) 41,548. cela displayed features characteristic of an end ...19979390455
overproduction of beta-glucosidase in active form by an escherichia coli system coexpressing the chaperonin groel/es.beta-glucosidase from cellvibrio gilvus was successfully overproduced in soluble form in escherichia coli, with the coexpression of groel/es. without the groel/es protein, the beta-glucosidase overexpressed in e. coli constituted a huge amount (80%) of the total cellular protein, but was localized in the insoluble fraction, and little activity was detected in the soluble fraction. coexpression of the e. coli groel/es had a drastic impact on the proper folding of the beta-glucosidase; 20% of the ...19989485593
natural cellulose fibers: heterogeneous acetylation kinetics and biodegradation behavior.steam-exploded fibers from flax (linum usitatissimum) are heterogeneously acetylated using acetic anhydride and sulfuric acid as catalyst, with the aim to modify the surface properties without changing fiber structure and morphology. the acetylation reaction follows first-order kinetics up to a reaction time that depends on catalyst concentration (15 h when using 0.4 vol % of h(2)so(4) or 50 h with 0.1 vol %). the fibers undergo no structural and/or morphological changes under either reaction co ...200111749209
cloning and sequencing of the beta-glucosidase gene from acetobacter xylinum atcc 23769.the beta-glucosidase gene (bglxa) was cloned from the genomic dna of acetobacter xylinum atcc 23769 and its nucleotide sequence (2200 bp) was determined. this bglxa gene was present downstream of the cellulose synthase operon and coded for a polypeptide of molecular mass 79 kda. the overexpression of the beta-glucosidase in a. xylinum caused a tenfold increase in activity compared to the wild-type strain. in addition, the action pattern of the enzyme was identified as g3ase activity. the deduced ...200111853314
identification of tandemly repeated type vi cellulose-binding domains in an endoglucanase from the aerobic soil bacterium cellvibrio mixtus.cellulose-binding domains (cbd) play a pivotal role during plant cell wall hydrolysis by cellulases and xylanases from aerobic soil bacteria. recently we have reported the molecular characterisation of a single-domain endoglucanase from cellvibrio mixtus, suggesting that some cellulases produced by this aerobic bacterium preferentially hydrolyse soluble cellulosic substrates. here we describe the complete nucleotide sequence of a second cellulase gene, celb, from the soil bacterium c. mixtus. it ...19989650253
a novel cellvibrio mixtus family 10 xylanase that is both intracellular and expressed under non-inducing conditions.hydrolysis of the plant cell wall polysaccharides cellulose and xylan requires the synergistic interaction of a repertoire of extracellular enzymes. recently, evidence has emerged that anaerobic bacteria can synthesize high levels of periplasmic xylanases which may be involved in the hydrolysis of small xylo-oligosaccharides absorbed by the micro-organism. cellvibrio mixtus, a saprophytic aerobic soil bacterium that is highly active against plant cell wall polysaccharides, was shown to express i ...200010931900
isolation and identification of cellulolytic bacteria involved in the degradation of natural cellulosic fibres.in search for bacterial cultures that are able to rapidly degrade cellulosic plant fibres in vitro, 77 cellulolytic strains were isolated from belgian and czech soils after enrichment on flax or sisal fibres as sole sources of carbon. the strains were characterized using fatty acid analysis, and 74 strains were grouped into three major clusters by numerical analysis. the first major cluster contained cellulomonas strains. within this cluster three subclusters could be delineated by principal com ...200010930083
kinetic studies of a recombinant cellobiose phosphorylase (cbp) of the clostridium thermocellum ym4 strain expressed in escherichia coli.a cellobiose phosphorylase (cbp) cloned from the clostridium thermocellum ym4 strain was purified to homogeneity, and the reaction mechanisms of both the phosphorolytic and synthetic reactions were studied in detail. the enzyme reaction proceeded via an ordered bi bi mechanism, in which p(i) bound to the enzyme prior to d-cellobiose and then g 1-p was released after d-glucose. the order of substrate binding was different from that of cbp from cellvibrio gilvus, which bound to cellobiose prior to ...200212153715
cellvibrio japonicus alpha-l-arabinanase 43a has a novel five-blade beta-propeller fold.cellvibrio japonicus arabinanase arb43a hydrolyzes the alpha-1,5-linked l-arabinofuranoside backbone of plant cell wall arabinans. the three-dimensional structure of arb43a, determined at 1.9 a resolution, reveals a five-bladed beta-propeller fold. arb43a is the first enzyme known to display this topology. a long v-shaped surface groove, partially enclosed at one end, forms a single extended substrate-binding surface across the face of the propeller. three carboxylates deep in the active site gr ...200212198486
the modular architecture of cellvibrio japonicus mannanases in glycoside hydrolase families 5 and 26 points to differences in their role in mannan degradation.beta-1,4-mannanases (mannanases), which hydrolyse mannans and glucomannans, are located in glycoside hydrolase families (ghs) 5 and 26. to investigate whether there are fundamental differences in the molecular architecture and biochemical properties of gh5 and gh26 mannanases, four genes encoding these enzymes were isolated from cellvibrio japonicus and the encoded glycoside hydrolases were characterized. the four genes, man5a, man5b, man5c and man26b, encode the mannanases man5a, man5b, man5c a ...200312523937
the membrane-bound alpha-glucuronidase from pseudomonas cellulosa hydrolyzes 4-o-methyl-d-glucuronoxylooligosaccharides but not 4-o-methyl-d-glucuronoxylan.the microbial degradation of xylan is a key biological process. hardwood 4-o-methyl-d-glucuronoxylans are extensively decorated with 4-o-methyl-d-glucuronic acid, which is cleaved from the polysaccharides by alpha-glucuronidases. in this report we describe the primary structures of the alpha-glucuronidase from cellvibrio mixtus (c. mixtus glca67a) and the alpha-glucuronidase from pseudomonas cellulosa (p. cellulosa glca67a) and characterize p. cellulosa glca67a. the primary structures of c. mixt ...200212169619
convergent evolution sheds light on the anti-beta -elimination mechanism common to family 1 and 10 polysaccharide lyases.enzyme-catalyzed beta-elimination of sugar uronic acids, exemplified by the degradation of plant cell wall pectins, plays an important role in a wide spectrum of biological processes ranging from the recycling of plant biomass through to pathogen virulence. the three-dimensional crystal structure of the catalytic module of a "family pl-10" polysaccharide lyase, pel10acm from cellvibrio japonicus, solved at a resolution of 1.3 a, reveals a new polysaccharide lyase fold and is the first example of ...200212221284
enhancement of transglycosylation activity by construction of chimeras between mesophilic and thermophilic beta-glucosidase.the family 3 beta-glucosidase from thermotoga maritima is a highly thermostable enzyme (85 degrees c) that displays transglycosylation activity. in contrast, the beta-glucosidase from cellvibrio gilvus is mesophilic (35 degrees c) and displays no such transglycosylation activity. both enzymes consist of two domains, an n-terminal and a c-terminal domain, and the amino acid identities between the two enzymes in these domains are 32.4 and 36.4%, respectively. in an attempt to identify the molecula ...200212392722
the alpha-glucuronidase, glca67a, of cellvibrio japonicus utilizes the carboxylate and methyl groups of aldobiouronic acid as important substrate recognition determinants.alpha-glucuronidases are key components of the ensemble of enzymes that degrade the plant cell wall. they hydrolyze the alpha1,2-glycosidic bond between 4-o-methyl-d-glucuronic acid (4-o-meglca) and the xylan or xylooligosaccharide backbone. here we report the crystal structure of an inactive mutant (e292a) of the alpha-glucuronidase, glca67a, from cellvibrio japonicus in complex with its substrate. the data show that the 4-o-methyl group of the substrate is accommodated within a hydrophobic she ...200312654910
molecular cloning, sequencing and expression of the gene encoding a novel chitinase a from a marine bacterium, pseudomonas sp pe2, and its domain structure.the pcha gene encoding chitinase a (pcha) from a pythium porphyrae cell-wall-degrading marine bacterium, pseudomonas sp. pe2, was cloned and characterized. the deduced pcha was a modular enzyme composed of an n-terminal signal peptide, a glycoside hydrolase family 18 catalytic domain that was responsible for the chitinase activity, the chitin-binding domains (chbds), and the carbohydrate-binding modules (cbm). the amino acid sequence of chbd(pcha) was highly conserved in the cbm family 12 that a ...200312655456
reclassification of 'pseudomonas fluorescens subsp. cellulosa' ncimb 10462 (ueda et al. 1952) as cellvibrio japonicus sp. nov. and revival of cellvibrio vulgaris sp. nov., nom. rev. and cellvibrio fulvus sp. nov., nom. rev.'pseudomonas fluorescens subsp. cellulosa' ncimb 10462 has been demonstrated by a polyphasic taxonomic approach to be a member of the genus cellvibrio. 16s rdna sequence analysis suggests that this is the only genus that could accept this specimen. the sequence is 95.5% similar to that of cellvibrio mixtus subsp. mixtus acm 2601t (the type strain of the type species of the genus), which is its closest relation. the genomic dna g + c content was determined to be 53.3 mol%, which is similar to the ...200312710603
purification and cellulolytic activity of cellvibrio. 195313063537
[the trace element requirements of cellvibrio and cytophaga]. 195313170582
taxonomic study of cellvibrio strains and description of cellvibrio ostraviensis sp. nov., cellvibrio fibrivorans sp. nov. and cellvibrio gandavensis sp. nov.thirty-one cellulolytic bacterial isolates from soils that were phenotypically very similar and phylogenetically highly related to cellvibrio strains were further characterized using a polyphasic taxonomic approach. by using repetitive extragenic palindromic dna-pcr fingerprinting, six different fingerprints could be recognized among the isolates. representative strains and four reference strains of the genus cellvibrio were used for dna-dna hybridization, which yielded eight dna hybridization g ...200312710614
importance of hydrophobic and polar residues in ligand binding in the family 15 carbohydrate-binding module from cellvibrio japonicus xyn10c.modular glycoside hydrolases that degrade the plant cell wall often contain noncatalytic carbohydrate-binding modules (cbms) that interact with specific polysaccharides within this complex macromolecule. cbms, by bringing the appended catalytic module into intimate and prolonged association with the substrate, increase the rate at which these enzymes are able to hydrolyze glycosidic bonds. recently, the crystal structure of the family 15 cbm (cbm15) from cellvibrio japonicus (formerly pseudomona ...200312899618
[significance of trace elements for cell vibrio and cytophaga species types]. 195613395442
[research on technic of isolation of cellvibrio]. 195213017148
metabolic basis for disaccharide preference in a cellvibrio. 195813610795
production of filterable particles by cellvibrio gilvus. 195913630875
disaccharide preference of an aerobic cellulolytic bacterium, cellvibrio gilvus n. sp. 195813610794
parallel induction of d-arabitol and d-sorbitol dehydrogenases.scolnick, edward m. (harvard medical school, boston, mass.) and edmund c. c. lin. parallel induction of d-arabitol and d-sorbitol dehydrogenases. j. bacteriol. 84:631-637. 1962.-two inducible diphosphopyridine nucleotide-linked dehydrogenases are described in a bacterium isolated from the soil, cellvibrio polyoltrophicus atcc 14774. the first enzyme catalyzes the dehydrogenation of d-arabitol to d-xylulose and d-mannitol to d-fructose. the data suggest that in vivo this enzyme has the dual funct ...196213992484
the complexity and mode of action of the cellulase system of cellvibrio gilvus. 196013834997
mode of action of a cellulase component from cellvibrio gilvus. 196314087368
green fluorescent pigment accumulated by a mutant of cellvibrio gilvus.love, samuel h. (bowman gray school of medicine, winston-salem, n.c.), and frank h. hulcher. green fluorescent pigment accumulated by a mutant of cellvibrio gilvus. j. bacteriol. 87:39-45. 1964.-a mutant of cellvibrio gilvus, designated strain 139a, liberated a green, fluorescent pigment into the surrounding culture medium. a study of the factors which affected the accumulation of this pigment led to the development of a chemically defined medium which supported maximal pigment accumulation in a ...196414102871
effect of water extracts of carob pods, tannic acid, and their derivatives on the morphology and growth of microorganisms.the effect of aqueous extracts of carob (ceratonia siliqua) pods, gallotannic acid, gallic acid, and catechol on several microorganisms was studied. carob pod extract and tannic acid showed a strong antimicrobial activity toward some cellulolytic bacteria. on the basis of tannin content, to which antimicrobial effect was related, carob pod extracts inhibited cellvibrio fulvus and clostridium cellulosolvens at 15 mug/ml, sporocytophaga myxococcoides at 45 mug/ml, and bacillus subtilis at 75 mug/m ...196414170956
structural and biochemical analysis of cellvibrio japonicus xylanase 10c: how variation in substrate-binding cleft influences the catalytic profile of family gh-10 xylanases.microbial degradation of the plant cell wall is the primary mechanism by which carbon is utilized in the biosphere. the hydrolysis of xylan, by endo-beta-1,4-xylanases (xylanases), is one of the key reactions in this process. although amino acid sequence variations are evident in the substrate binding cleft of "family gh10" xylanases (see afmb.cnrs-mrs.fr/cazy/), their biochemical significance is unclear. the cellvibrio japonicus gh10 xylanase cjxyn10c is a bi-modular enzyme comprising a gh10 ca ...200414670951
utilization of cellulose oligosaccharides by cellvibrio gilvus.schafer, marion l. (virginia polytechnic institute, blacksburg), and kendall w. king. utilization of cellulose oligosaccharides by cellvibrio gilvus. j. bacteriol. 89:113-116. 1965.-the hypothesis that oligosaccharides of the cellulose polymer series can be absorbed by cellulolytic bacteria, prior to hydrolysis to the level of glucose or cellobiose, has been tested. resting-cell suspensions of cellvibrio gilvus removed oligosaccharides of one to six monomer units from solution at a rate providin ...196514255649
metabolic nonequivalence of the two glucose moieties of cellobiose in cellvibrio gilvus.swisher, elizabeth j. (virginia polytechnic institute, blacksburg), waldemar o. storvick, and kendall w. king. metabolic nonequivalence of the two glucose moieties of cellobiose in cellvibrio gilvus. j. bacteriol. 88:817-820. 1964.-cellobiose was synthesized in 40% yield with uniform c(14) labeling in the reducing glucose moiety and no label in the nonreducing glucosyl. resting-cell suspensions of cellvibrio gilvus respiring the labeled cellobiose derived approximately 80% of their respiratory c ...196414219041
the family 6 carbohydrate binding module cmcbm6-2 contains two ligand-binding sites with distinct specificities.the microbial degradation of the plant cell wall is an important biological process, representing a major component of the carbon cycle. enzymes that mediate the hydrolysis of this composite structure are modular proteins that contain non-catalytic carbohydrate binding modules (cbms) that enhance catalytic activity. cbms are grouped into sequence-based families, and in a previous study we showed that a family 6 cbm (cbm6) that interacts with xylan contains two potential ligand binding clefts, de ...200415004011
x4 modules represent a new family of carbohydrate-binding modules that display novel properties.the hydrolysis of the plant cell wall by microbial glycoside hydrolases and esterases is the primary mechanism by which stored organic carbon is utilized in the biosphere, and thus these enzymes are of considerable biological and industrial importance. plant cell wall-degrading enzymes in general display a modular architecture comprising catalytic and non-catalytic modules. the x4 modules in glycoside hydrolases represent a large family of non-catalytic modules whose function is unknown. here we ...200415004012
three acidic residues are at the active site of a beta-propeller architecture in glycoside hydrolase families 32, 43, 62, and 68.multiple-sequence alignment of glycoside hydrolase (gh) families 32, 43, 62, and 68 revealed three conserved blocks, each containing an acidic residue at an equivalent position in all the enzymes. a detailed analysis of the site-directed mutations so far performed on invertases (gh32), arabinanases (gh43), and bacterial fructosyltransferases (gh68) indicated a direct implication of the conserved residues asp/glu (block i), asp (block ii), and glu (block iii) in substrate binding and hydrolysis. ...200414747991
inactivated enzymes as probes of the structure of arabinoxylans as observed by atomic force microscopy.the complex structures of water-soluble wheat arabinoxylans have been mapped along individual molecules, and within populations, using the visualisation of the binding of inactivated enzymes by atomic force microscopy (afm). it was demonstrated that site-directed mutagenesis (sdm) can be used to produce inactive enzymes as structural probes. for the sdm mutants afm has been used to compare the binding of different xylanases to arabinoxylans. xylanase mutant e386a, derived from the xyn11a enzyme ...200415013394
biodegradation of chemically modified flax fibers in soil and in vitro with selected bacteria.the extent and rate of degradation of flax (linum usitatissimum) fibers, both in the native state and after surface chemical modification (acetylation or poly(ethylene glycol), peg, grafting), was investigated under laboratory conditions in two different biodegrading environments. degradation of the fibers under aerobic conditions by the action of the microorganisms present in soil is assessed with the astm 5988-96 method by monitoring carbon dioxide evolution. in vitro biodegradation experiment ...200415003026
the mechanisms by which family 10 glycoside hydrolases bind decorated substrates.endo-beta-1,4-xylanases (xylanases), which cleave beta-1,4 glycosidic bonds in the xylan backbone, are important components of the repertoire of enzymes that catalyze plant cell wall degradation. the mechanism by which these enzymes are able to hydrolyze a range of decorated xylans remains unclear. here we reveal the three-dimensional structure, determined by x-ray crystallography, and the catalytic properties of the cellvibrio mixtus enzyme xyn10b (cmxyn10b), the most active gh10 xylanase descr ...200414668328
insights into the molecular determinants of substrate specificity in glycoside hydrolase family 5 revealed by the crystal structure and kinetics of cellvibrio mixtus mannosidase 5a.the enzymatic hydrolysis of the glycosidic bond is central to numerous biological processes. glycoside hydrolases, which catalyze these reactions, are grouped into families based on primary sequence similarities. one of the largest glycoside hydrolase families is glycoside hydrolase family 5 (gh5), which contains primarily endo-acting enzymes that hydrolyze beta-mannans and beta-glucans. here we report the cloning, characterization, and three-dimensional structure of the cellvibrio mixtus gh5 be ...200415014076
the crystal structure of the family 6 carbohydrate binding module from cellvibrio mixtus endoglucanase 5a in complex with oligosaccharides reveals two distinct binding sites with different ligand specificities.glycoside hydrolases that release fixed carbon from the plant cell wall are of considerable biological and industrial importance. these hydrolases contain non-catalytic carbohydrate binding modules (cbms) that, by bringing the appended catalytic domain into intimate association with its insoluble substrate, greatly potentiate catalysis. family 6 cbms (cbm6) are highly unusual because they contain two distinct clefts (cleft a and cleft b) that potentially can function as binding sites. henshaw et ...200415010454
azospirillum irakense pectate lyase displays a toroidal fold.the three-dimensional structure of azospirillum irakense pectate lyase (pela) has been determined at a resolution of 2.65 a. the crystals are hexagonal, belonging to space group p6(5)22, with unit-cell parameters a = b = 85.37, c = 231.32 angstroms. phase information was derived from a multiple-wavelength anomalous dispersion (mad) experiment using a hg derivative. refinement of the model converged to rcryst = 20.08% and rfree = 25.87%. the overall structure of pela does not adopt the characteri ...200415159558
crystallization and preliminary x-ray diffraction analysis of a thermostable endo-1,5-alpha-l-arabinanase from bacillus thermodenitrificans ts-3.a thermostable endo-1,5-alpha-l-arabinanase abn-ts from bacillus thermodenitrificans ts-3 with a molecular weight of 35 kda was crystallized by the hanging-drop vapour-diffusion method using sodium citrate as a precipitant. the crystals were loop-mounted in a cryoprotectant solution containing 28%(w/v) sucrose and 1 m sodium citrate ph 6.0 and flash-cooled. sucrose was selected as the most suitable cryoprotectant. the crystal belonged to the orthorhombic space group p2(1)2(1)2(1), with unit-cell ...200415159584
tailored catalysts for plant cell-wall degradation: redesigning the exo/endo preference of cellvibrio japonicus arabinanase 43a.enzymes acting on polymeric substrates are frequently classified as exo or endo, reflecting their preference for, or ignorance of, polymer chain ends. most biotechnological applications, especially in the field of polysaccharide degradation, require either endo- or exo-acting hydrolases, or they harness the essential synergy between these two modes of action. here, we have used genomic data in tandem with structure to modify, radically, the chain-end specificity of the cellvibrio japonicus exo-a ...200515708971
structure of a mannan-specific family 35 carbohydrate-binding module: evidence for significant conformational changes upon ligand binding.enzymes that digest plant cell wall polysaccharides generally contain non-catalytic, carbohydrate-binding modules (cbms) that function by attaching the enzyme to the substrate, potentiating catalytic activity. here, we present the first structure of a family 35 cbm, derived from the cellvibrio japonicus beta-1,4-mannanase man5c. the nmr structure has been determined for both the free protein and the protein bound to mannopentaose. the data show that the protein displays a typical beta-jelly-roll ...200515740741
crystallization and preliminary x-ray analysis of cellobiose phosphorylase from cellvibrio gilvus.a recombinant cellobiose phosphorylase from cellvibrio gilvus has been prepared and crystallized by the sitting-drop vapour-diffusion method using 10 mg ml(-1) purified enzyme, 1.5 m ammonium sulfate, 0.1 m mes buffer ph 7.0 and 5 mm glucose. a suitable crystal was obtained after 10 d incubation at 298 k. the crystal belongs to space group p2(1), with unit-cell parameters a = 84.77, b = 98.31, c = 104.04 a, beta = 102.73 degrees. x-ray diffraction data to 2.1 a resolution have been collected at ...200415388938
effect of dimer dissociation on activity and thermostability of the alpha-glucuronidase from geobacillus stearothermophilus: dissecting the different oligomeric forms of family 67 glycoside hydrolases.the oligomeric organization of enzymes plays an important role in many biological processes, such as allosteric regulation, conformational stability and thermal stability. alpha-glucuronidases are family 67 glycosidases that cleave the alpha-1,2-glycosidic bond between 4-o-methyl-d-glucuronic acid and xylose units as part of an array of hemicellulose-hydrolyzing enzymes. currently, two crystal structures of alpha-glucuronidases are available, those from geobacillus stearothermophilus (agua) and ...200415466046
the use of forced protein evolution to investigate and improve stability of family 10 xylanases. the production of ca2+-independent stable xylanases.metal ions such as calcium often play a key role in protein thermostability. the inclusion of metal ions in industrial processes is, however, problematic. thus, the evolution of enzymes that display enhanced stability, which is not reliant on divalent metals, is an important biotechnological goal. here we have used forced protein evolution to interrogate whether the stabilizing effect of calcium in an industrially relevant enzyme can be replaced with amino acid substitutions. our study has focus ...200415452124
the structure and characterization of a modular endo-beta-1,4-mannanase from cellulomonas fimi.the endo-beta-1,4-mannanase from the soil bacterium cellulomonas fimi is a modular plant cell wall degrading enzyme involved in the hydrolysis of the backbone of mannan, one of the most abundant polysaccharides of the hemicellulosic network in the plant cell wall. the crystal structure of a recombinant truncated endo-beta-1,4-mannanase from c. fimi (cfman26a-50k) was determined by x-ray crystallography to 2.25 a resolution using the molecular replacement technique. the overall structure of the e ...200516171384
sequence of the gene for a high-alkaline mannanase from an alkaliphilic bacillus sp. strain jamb-750, its expression in bacillus subtilis and characterization of the recombinant enzyme.a novel alkaline mannanase man26a has been found in the culture of an alkaliphilic bacillus sp. strain jamb-750 and the optimal ph for the mannanase activity of the enzyme was around ph 10 (j biol macromol 4: 67-74, 2004). this optimal ph is the highest among those of the mannanases reported to date. the gene man26a coding the enzyme was cloned from the genomic dna of strain jamb-750 and sequenced. it encodes a protein of 997 amino acids including a signal peptide. the n-terminal half (glu27-val ...200515999223
screening of novel cellulose-degrading bacterium and its application to denitrification of groundwater.to establish an environmentally friendly groundwater bioremediation process using a cellulose carrier combined with cellulose-utilizing, denitrifying microorganisms, a novel psychrophilic bacterium, designated cl-5, which can degrade a commercial-based cellulose carrier as the sole carbon source, was screened. since the denitrification capability of cl-5 is low, complex microbial systems were constructed together with other denitrifying bacteria designated nr-1 and nr-2 that were also isolated f ...200516233813
some properties of cellulolytic cellvibrio strains from polluted water. 196816349814
identification and specific detection of a novel pseudomonadaceae cluster associated with soils from winter wheat plots of a long-term agricultural field experiment.the genus pseudomonas (sensu stricto) represents a group of microorganisms directly involved in functions conferring plant health. we performed a study in the dok long-term agricultural field experiment on the basis of previously published pseudomonas-selective pcr primers in order to investigate the community structure of the microbial groups defined by the target range of these primers. three different agricultural management systems, i.e., conventional, biodynamic, and bio-organic, along with ...200616391022
bacteria associated with spores of the arbuscular mycorrhizal fungi glomus geosporum and glomus constrictum.spores of the arbuscular mycorrhizal fungi (amf) glomus geosporum and glomus constrictum were harvested from single-spore-derived pot cultures with either plantago lanceolata or hieracium pilosella as host plants. pcr-denaturing gradient gel electrophoresis analysis revealed that the bacterial communities associated with the spores depended more on amf than host plant identity. the composition of the bacterial populations linked to the spores could be predominantly influenced by a specific spore ...200516269696
structural insight into the ligand specificity of a thermostable family 51 arabinofuranosidase, araf51, from clostridium thermocellum.the digestion of the plant cell wall requires the concerted action of a diverse repertoire of enzyme activities. an important component of these hydrolase consortia are arabinofuranosidases, which release l-arabinofuranose moieties from a range of plant structural polysaccharides. the anaerobic bacterium clostridium thermocellum, a highly efficient plant cell wall degrader, possesses a single alpha-l-arabinofuranosidase (ec 3.2.1.55), ctaraf51a, located in gh51 (glycoside hydrolase family 51). t ...200616336192
reaction on d-glucal by an inverting phosphorylase to synthesize derivatives of 2-deoxy-beta-d-arabino-hexopyranosyl-(1-->4)-d-glucose (2ii-deoxycellobiose).four derivatives of 2(ii)-deoxycellobiose were synthesized from d-glucal and acceptor sugars (d-glucose, d-xylose, d-mannose, and 2-deoxy-d-arabino-hexose) using a cellobiose phosphorylase from cellvibrio gilvus. the enzyme was found to be an effective catalyst to synthesize the beta-(1-->4) linkage of 2-deoxy-d-arabino-hexopyranoside. the acceptor specificity for the d-glucal reaction was identical to that for the alpha-d-glucose 1-phosphate reaction, but the activity of d-glucal was approximat ...200616430877
cloning of a gene cluster from cellvibrio mixtus which codes for cellulase, chitinase, amylase, and pectinase.the soil isolate cellvibrio mixtus uqm2294 degraded a variety of polysaccharides including microcrystalline cellulose. among 6,000 cosmid clones carrying c. mixtus dna, constructed in escherichia coli with phc79, 50 expressed the ability to degrade one or more of the following substrates: carboxymethyl cellulose, chitin, pectin (polygalacturonic acid), cellobiose, and starch. these degradative genes are encoded in a single 94.1-kilobase segment of the c. mixtus genome; a preliminary order of the ...198616347240
characterization of a metagenome-derived halotolerant cellulase.metagenomes of uncultured microorganisms represent a sheer unlimited resource for discovery of novel biocatalysts. here, we report on the biochemical characterisation of a novel, soil metagenome-derived cellulase (endoglucanase), cel5a. the deduced amino acid sequence of cel5a was similar to a family 5, single domain cellulase with no distinct cellulose binding domain from cellvibrio mixtus. the 1092bp orf encoding cel5a was overexpressed in escherichia coli and the corresponding 42.1 kda protei ...200616584799
structural dissection of the reaction mechanism of cellobiose phosphorylase.cellobiose phosphorylase, a member of the glycoside hydrolase family 94, catalyses the reversible phosphorolysis of cellobiose into alpha-d-glucose 1-phosphate and d-glucose with inversion of the anomeric configuration. the substrate specificity and reaction mechanism of cellobiose phosphorylase from cellvibrio gilvus have been investigated in detail. we have determined the crystal structure of the glucose-sulphate and glucose-phosphate complexes of this enzyme at a maximal resolution of 2.0 a ( ...200616646954
probing the structural basis for the difference in thermostability displayed by family 10 xylanases.thermostability is an important property of industrially significant hydrolytic enzymes: understanding the structural basis for this attribute will underpin the future biotechnological exploitation of these biocatalysts. the cellvibrio family 10 (gh10) xylanases display considerable sequence identity but exhibit significant differences in thermostability; thus, these enzymes represent excellent models to examine the structural basis for the variation in stability displayed by these glycoside hyd ...200616762367
computational analyses of the conformational itinerary along the reaction pathway of gh94 cellobiose phosphorylase.gh94 cellobiose phosphorylase (cbp) catalyzes the phosphorolysis of cellobiose into alpha-d-glucose 1-phosphate (g1p) and d-glucose with inversion of anomeric configuration. the complex crystal structure of cbp from cellvibrio gilvus had previously been determined; glycerol, glucose, and phosphate are bound to subsites -1, +1, and the anion binding site, respectively. we performed computational analyses to elucidate the conformational itinerary along the reaction pathway of this enzyme. autodock ...200818346721
simiduia agarivorans gen. nov., sp. nov., a marine, agarolytic bacterium isolated from shallow coastal water from keelung, taiwan.a gram-negative, heterotrophic, agarolytic, marine bacterium, designated strain sa1t, was isolated from a seawater sample collected in the shallow coastal region of keelung, taiwan. cells were straight to slightly curved rods. nearly all of the cells were non-motile and non-flagellated during the exponential phase of growth in broth cultures; a few cells (<1 %) were motile and were considered to have monotrichous flagella. the isolate required nacl for growth and grew optimally at 30-35 degrees ...200818398190
insights into plant cell wall degradation from the genome sequence of the soil bacterium cellvibrio japonicus.the plant cell wall, which consists of a highly complex array of interconnecting polysaccharides, is the most abundant source of organic carbon in the biosphere. microorganisms that degrade the plant cell wall synthesize an extensive portfolio of hydrolytic enzymes that display highly complex molecular architectures. to unravel the intricate repertoire of plant cell wall-degrading enzymes synthesized by the saprophytic soil bacterium cellvibrio japonicus, we sequenced and analyzed its genome, wh ...200818556790
phylogenetic and metabolic bacterial diversity of phragmites australis periphyton communities in two hungarian soda ponds.bacterial diversity of reed (phragmites australis) periphyton communities of kelemen-szék and nagy-vadas (two hungarian soda ponds) was investigated using molecular cloning and cultivation-based techniques. the majority of the 80 kelemen-szék and 72 nagy-vadas bacterial isolates proved to be moderately halophilic and alkaliphilic. a great proportion of the isolates showed phosphatase and urease activity, utilized aesculin, citrate and certain biopolymers (e.g., gelatine and tween 80). partial 16 ...200818679563
the cellvibrio japonicus mannanase cjman26c displays a unique exo-mode of action that is conferred by subtle changes to the distal region of the active site.the microbial degradation of the plant cell wall is a pivotal biological process that is of increasing industrial significance. one of the major plant structural polysaccharides is mannan, a beta-1,4-linked d-mannose polymer, which is hydrolyzed by endo- and exo-acting mannanases. the mechanisms by which the exo-acting enzymes target the chain ends of mannan and how galactose decorations influence activity are poorly understood. here we report the crystal structure and biochemical properties of ...200818799462
dasania marina gen. nov., sp. nov., of the order pseudomonadales, isolated from arctic marine sediment.an obligately aerobic bacterium, strain kopri 20902t, was isolated from a marine sediment in ny-arlesund, spitsbergen islands, norway. cells were irregular rods and motile with polar monotrichous flagellum. the optimum growth temperature was 17-22 degrees . cells grew best in ph 7.0-10.0 and 3-4% sea salts (corresponding to 2.3-3.1% nacl). the novel strain required ca2+ or mg2+ in addition to nacl for growth. sequence analysis of 16s rrna gene revealed that the arctic isolate is distantly relate ...200718176532
the active site of a carbohydrate esterase displays divergent catalytic and noncatalytic binding functions.multifunctional proteins, which play a critical role in many biological processes, have typically evolved through the recruitment of different domains that have the required functional diversity. thus the different activities displayed by these proteins are mediated by spatially distinct domains, consistent with the specific chemical requirements of each activity. indeed, current evolutionary theory argues that the colocalization of diverse activities within an enzyme is likely to be a rare even ...200919338387
understanding how diverse beta-mannanases recognize heterogeneous substrates.the mechanism by which polysaccharide-hydrolyzing enzymes manifest specificity toward heterogeneous substrates, in which the sequence of sugars is variable, is unclear. an excellent example of such heterogeneity is provided by the plant structural polysaccharide glucomannan, which comprises a backbone of beta-1,4-linked glucose and mannose units. beta-mannanases, located in glycoside hydrolase (gh) families 5 and 26, hydrolyze glucomannan by cleaving the glycosidic bond of mannosides at the -1 s ...200919441796
factor g utilizes a carbohydrate-binding cleft that is conserved between horseshoe crab and bacteria for the recognition of beta-1,3-d-glucans.in the horseshoe crab, the recognition of beta-1,3-d-glucans by factor g triggers hemolymph coagulation. factor g contains a domain of two tandem xylanase z-like modules (z1-z2), each of which recognizes beta-1,3-d-glucans. to gain an insight into the recognition of beta-1,3-d-glucans from a structural view point, recombinants of z1-z2, the c-terminal module z2, z2 with a cys to ala substitution (z2a), and its tandem repeat z2a-z2a were characterized. z2 and z1-z2, but not z2a and z2a-z2a, forme ...200919710471
family 6 carbohydrate-binding modules display multiple beta1,3-linked glucan-specific binding interfaces.noncatalytic carbohydrate-binding modules (cbms), which are found in a variety of carbohydrate-degrading enzymes, have been grouped into sequence-based families. cbms, by recruiting their appended enzymes onto the surface of the target substrate, potentiate catalysis particularly against insoluble substrates. family 6 cbms (cbm6s) display unusual properties in that they present two potential ligand-binding sites termed clefts a and b, respectively. cleft b is located on the concave surface of th ...200919751219
galactomannan hydrolysis and mannose metabolism in cellvibrio mixtus.galactomannan hydrolysis results from the concerted action of microbial endo-mannanases, manosidases and alpha-galactosidases and is a mechanism of intrinsic biological importance. here we report the identification of a gene cluster in the aerobic soil bacterium cellvibrio mixtus encoding enzymes involved in the degradation of this polymeric substrate. the family 27 alpha-galactosidase, termed cmaga27a, preferentially hydrolyse galactose containing polysaccharides. in addition, we have character ...200616842369
analysis of a change in bacterial community in different environments with addition of chitin or chitosan.the temporal changes of a bacterial community in soil with chitin or chitosan added were analyzed by pcr-denaturing gradient gel electrophoresis (dgge) targeting the 16s rrna gene using total dnas prepared from the community. band patterns of pcr-dgge confirmed that 31 species become predominant after the addition of chitin or chitosan. the determination of the nucleotide sequences of the bands of the 31 species indicated that 20 species belonged to the division proteobacteria, and that the genu ...201020347770
rational engineering of mannosyl binding in the distal glycone subsites of cellulomonas fimi endo-beta-1,4-mannanase: mannosyl binding promoted at subsite -2 and demoted at subsite -3.to date, rational redesign of glycosidase active-site clefts has been mainly limited to the removal of essential functionalities rather than their introduction. the glycoside hydrolase family 26 endo-beta-1,4-mannanase from the soil bacterium cellulomonas fimi depolymerizes various abundant plant mannans. on the basis of differences in the structures and hydrolytic action patterns of this wild-type (but recombinantly expressed) enzyme and a homologous mannanase from cellvibrio japonicus, two non ...201020426480
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