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...

A [Fe]-hydrogenase gene (hydA) was cloned from Clostridium paraputrificum M-21 in Escherichia coli using a conserved DNA sequence of clostridial hydrogenase genes amplified by PCR as the probe. The hydA consisted an open reading frame 1749 bp encoding 582 amino acids with estimated molecular mass 64,560 Da. It ligated into shuttle vector, pJIR751, originally constructed for perfringens and E. coli, expressed C. paraputrificum. Hydrogen gas productivity recombinant increased up to 1.7-fold...

Two manganese peroxidases (MnPs), MnP1 and MnP2, a laccase, Lac1, were purified from Trametes polyzona KU-RNW027. Both MnPs showed high stability in organic solvents which triggered their activities. Metal ions activated both at certain concentrations. The two played important roles dye degradation pharmaceutical products deactivation redox mediator-free system. They completely degraded Remazol brilliant blue (25 mg/L) 10-30 min activities to navy yellow, while Lac1 could remove 75% of red....

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...

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...

The arsenic resistance (ars) operon from plasmid pKW301 of Acidiphilium multivorum AIU 301 was cloned and sequenced. This DNA sequence contains five genes in the following order: arsR, arsD, arsA, arsB, arsC. predicted amino acid sequences all gene products are homologous to ars Escherichia coli R773 IncN R46. A. conferred arsenate arsenite on E. coli. Expression with bacteriophage T7 RNA polymerase-promoter system allowed overexpress ArsD, ArsA, ArsC but not ArsR or ArsB. apparent molecular...

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...

Complete utilization of carbohydrate fractions is one the prerequisites for obtaining economically favorable lignocellulosic biomass conversion. This study shows that xylan in untreated rice straw was saccharified to xylose step without chemical pretreatment, yielding 58.2% theoretically maximum value by Paenibacillus curdlanolyticus B-6 PcAxy43A, a weak lignin-binding trifunctional xylanolytic enzyme, endoxylanase/β-xylosidase/arabinoxylan arabinofuranohydrolase. Moreover, yield from...

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...