期刊
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
卷 65, 期 3, 页码 548-554出版社
TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.65.548
关键词
Clostridium thermocellum; mannanase; cellulosome
The man26B gene of Clostridium thermocellum strain F1 was found in pKS305, which had been selected as a recombinant plasmid conferring endoglucanase activity on Escherichia coli, The open reading frame of man26B consists of 1,773 nucleotides encoding a protein of 591 amino acids with a predicted molecular weight of 67,047, Man26B is a modular enzyme composed of an :V-terminal signal peptide and three domains in the following order: a mannan-binding domain, a family 26 mannanase domain, and a dockerin domain responsible for cellulosome assembly. We found that this gene was a homologue of the man26A gene of C, thermocellum strain YS but that there were insertion or deletion mutations that caused a frame-shift mutation affecting a stretch of 26 amino acids in the catalytic domain. Man26B devoid of the dockerin domain was constructed and purified from a recombinant E. coli, and its enzyme properties were examined. Immunological analysis indicated that Man26B was a catalytic component of the C. thermocellum Fl cellulosome.
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