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Volume 148, Issue 1

A newly described cellulosomal cellobiohydrolase, CelO, from : investigation of the exo-mode of hydrolysis, and binding capacity to crystalline cellulose

The GenBank accession number for the sequence determined in this work is AJ275975.

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Abstract

The sequence of the gene from F7 was determined. The gene product, cellulase CelO (Ct-Cel5F), had a modular structure consisting of a carbohydrate-binding module of the CBM3 family and a catalytic domain of the glycosyl hydrolase family 5. The presence of the dockerin module indicated that the enzyme was a component of the cellulosome complex. The thermostable recombinant gene product was active on cellodextrins, barley β-glucan, carboxymethylcellulose and insoluble cellulose. Cellobiose was the only product released from amorphic and crystalline cellulose, cellotetraose and higher cello-oligosaccharides, identifying CelO as a cellobiohydrolase. The cleavage pattern of nitrophenyl β-D-cellotetraoside, blockage of the hydrolysis of NaBH-reduced cellopentaose and the reduction in substrate viscosity suggested activity from the reducing end in a processive mode after making random cuts. Binding to insoluble, i.e. amorphous, and crystalline cellulose was mediated by the carbohydrate-binding module CBM3b, with a preference for the crystalline substrate.

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Keyword(s): carbohydrate-binding module CBM3 , CBM, carbohydrate-binding module , cellulase CelO , cellulosome , CMC, carboxymethylcellulose , GHF, glycosyl hydrolase family , PASC, phosphoric acid swollen cellulose , pNP, p-nitrophenyl and reducing end
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2002-01-01
2025-04-26
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/content/journal/micro/10.1099/00221287-148-1-247
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A newly described cellulosomal cellobiohydrolase, CelO, from Clostridium thermocellum: investigation of the exo-mode of hydrolysis, and binding capacity to crystalline celluloseThe GenBank accession number for the sequence determined in this work is AJ275975.
Microbiology 148, 247 (2002); https://doi.org/10.1099/00221287-148-1-247
/content/journal/micro/10.1099/00221287-148-1-247
/content/journal/micro/10.1099/00221287-148-1-247
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