ABSTRACT The Clostridium josui cipA and celD genes, encoding a scaffolding-like protein (CipA) putative cellulase (CelD), respectively, have been cloned sequenced. CipA, with an estimated molecular weight of 120,227, consists N-terminal signal peptide, cellulose-binding domain family III, six successive cohesin domains. architecture C. CipA is similar to those the scaffolding proteins reported so far, such as thermocellum cellulovorans CbpA, cellulolyticum CipC, but considerably smaller than...

The Clostridium paraputrificum chiB gene, encoding chitinase B (ChiB), consists of an open reading frame 2,493 nucleotides and encodes 831 amino acids with a deduced molecular weight 90,020. ChiB is modular enzyme composed family 18 catalytic domain responsible for activity, two reiterated domains unknown function, chitin-binding (CBD). are similar to the repeating units cadherin proteins but not fibronectin type III domains, therefore they referred as cadherin-like domains. was purified...

The cellulolytic complex was isolated from the culture supernatant of Ruminococcus albus strain F-40 grown on cellulose by a Sephacryl S-300HR column chromatography. molecular mass found to be larger than 1.5×106 Da. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated that contained at least 15 proteins with weights 40 kDa 250 kDa. Among them, 11 showed endoglucanase and/or xylanase activities zymograms. Immunological using an antiserum raised against...

ABSTRACT A large gene cluster for the Clostridium cellulovorans cellulosome has been cloned and sequenced upstream downstream of cbpA exgS genes (C.-C. Liu R. H. Doi, Gene 211:39–47, 1998). walking revealed that engL (Y. Tamaru J. Bacteriol. 182:244–247, 2000) was located cbpA-exgS genes. Further DNA sequencing this contains scaffolding protein CbpA, exoglucanase ExgS, several endoglucanases family 9, mannanase ManA, hydrophobic HbpA containing a surface layer homology domain (or cohesin)...

L-Galactono-γ-lactone dehydrogenase (EC 1.3.2.3, GLDHase) was partially purified from mitochondria of sweet potato tuberous roots over 600-fold on a specific activity basis, followed by purification the enzyme protein 56 kDa preparative SDS-PAGE. The absorption spectrum hydroxylapatite column-purified GLDHase showed peaks at 448 and 373 nm, suggesting presence flavin as prosthetic group. inhibited lycorine, an alkaloid which inhibits ascorbic acid biosynthesis in vivo. N-terminal partial...

The nucleotides of the xynA gene Clostridium stercorarium were sequenced. structural consists an open reading frame 1533 bp encoding 511 amino acids with M(r) 56,519. signal peptide cleavage site was identified by comparison N-terminal acid sequence enzyme produced a recombinant Escherichia coli. Xylanase A catalytic domain belonging to family G at N-terminus and two direct repeats about 90 short spacing C-terminus. Deletion analysis showed that repeated sequences responsible for binding...

In enzymatic saccharification of lignocellulosics, the access enzymes to exposed cellulose surfaces is a key initial step in triggering hydrolysis. However, knowledge structure-hydrolyzability relationship pretreated biomass still limited. Here we used fluorescent-labeled recombinant carbohydrate-binding modules (CBMs) from Clostridium josui as specific markers for crystalline (CjCBM3) and non-crystalline (CjCBM28) analyze complex wood tissues with NaOH, NaOH-Na(2)S (kraft pulping),...

The Clostridium thermocellum F1 celJ gene, encoding endoglucanase J (CelJ), consists of an open reading frame (ORF) 4,803 nucleotides and encodes a protein 1,601 amino acids with molecular weight 178,055. ORF was confirmed as by comparison the N-terminal sequence truncated CelJ derivative. is modular enzyme composed signal peptide six domains in following order: S-layer homology domain, domain unknown function (UD-1), subfamily E1 family docking another (UD-2). UD-1 has no significant...

The nucleotide sequence of the Clostridium thermocellum F1 xynC gene, which encodes xylanase XynC, consists 1,857 bp and a protein 619 amino acids with molecular weight 69,517. XynC contains typical N-terminal signal peptide 32 acid residues, followed by 165-amino-acid is homologous to thermostabilizing domain. Downstream this domain was family 10 catalytic glycosyl hydrolase. C terminus separated from short linker dockerin responsible for cellulosome assembly. XynC-II, enzyme purified...

ABSTRACT Clostridium thermocellum CelJ is a modular enzyme containing family 30 carbohydrate-binding module (CBM) and 9 catalytic at its N-terminal moiety. To investigate the functions of CBM module, truncated derivatives were constructed characterized. Isothermal titration calorimetric studies showed that association constants ( K ) polypeptide (CBM30) for binding cellopentaose cellohexaose 1.2 × 10 4 6.4 M −1 , respectively, CBM30 to these ligands enthalpically driven. Qualitative analyses...

Penicillum sp. 40, which can grow in an extremely acidic medium at pH 2.0 was screened from soil. This fungus produces xylanases when grown a containing xylan as sole carbon source. A major xylanase purified the culture supernatant of Penicillium 40 and designated XynA. The molecular mass XynA estimated to be 25,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. has optimum is stable 2.0-5.0. Western blot analysis using anit-XynA antibody showed that induced repressed glucose....

The Clostridium stercorarium xylanase Xyn10B is a modular enzyme comprising two thermostabilizing domains, family 10 catalytic domain of glycosyl hydrolases, 9 carbohydrate-binding module (CBM), and S-layer homologous (SLH) domains [Biosci. Biotechnol. Biochem., 63, 1596-1604 (1999)]. To investigate the role this CBM, we constructed derivatives compared their hydrolytic activity toward xylan some preparations plant cell walls; Xyn10BdeltaCBM consists only, Xyn10B-CBM comprises CBM. bound to...

Using dog food as a model of the organic waste, thermophilic composting was carried out for 14 days at fixed temperature 60 degrees C. The relationship between matter decomposition measured by CO2 evolution during bio-stabilization process and microbial succession expressed changes over time in restriction fragment length polymorphism (RFLP) patterns 16S rDNA sequences, micro-organisms associated with material also examined. rate peaked on day 3 gradually decreased until it became extremely...

Abstract BACKGROUND Total mixed ration ( TMR ) is widely used for dairy cattle and needs to be prepared daily because it deteriorates rapidly. Ensiling allows preservation saves labour at the farm; however, silage fermentation may influence various nutritional components. The objectives of this study were evaluate changes in vitro rumen that was stored different temperatures durations on a laboratory scale comparison with those typical before ensiling. RESULTS No distinct crude protein CP ),...

The cellulosome components are assembled into the complex by interaction between one of repeated cohesin domains a scaffolding protein and dockerin domain an enzyme component. We prepared five recombinant polypeptides Clostridium thermocellum CipA, two C. Xyn11A Xyn10C, four josui Aga27A Cel8A, qualitatively quantitatively examined cohesin-dockerin interactions within josui, respectively, species specificity these bacteria. Surface plasmon resonance (SPR) analysis indicated that there was...

The crystal structure of Cel44A, which is one the enzymatic components cellulosome Clostridium thermocellum, was solved at a resolution 0.96 A. This enzyme belongs to glycoside hydrolase family (GH family) 44. reveals that Cel44A consists TIM-like barrel domain and beta-sandwich domain. wild-type E186Q mutant structures complexed with substrates suggest two glutamic acid residues, Glu(186) Glu(359), are active residues enzyme. Biochemical experiments were performed confirm this idea....

ABSTRACT In general, cellulases and hemicellulases are modular enzymes in which the catalytic domain is appended to one or more noncatalytic carbohydrate binding modules (CBMs). CBMs, by concentrating parental enzyme at their target polysaccharide, increase capacity of module bind substrate, leading a potentiation catalysis. Clostridium thermocellum hypothetical protein Cthe_0821, defined here as C. Man5A, comprising an N-terminal signal peptide, family 5 glycoside hydrolase (GH5) module, 32...