Sequence of the Clostridium thermocellum mannanase gene man26B and characterization of the translated product

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 mol...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2001-03, Vol.65 (3), p.548-554
Main Authors: Kurokawa, J. (Mie Univ., Tsu (Japan). Faculty of Bioresources), Hemjinda, E, Arai, T, Karita, S, Kimura, T, Sakka, K, Ohmiya, K
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
beta-Mannosidase
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
cellulosome
Clostridium - enzymology
Clostridium - genetics
CLOSTRIDIUM THERMOCELLUM
DNA, Bacterial
ENZYMIC ACTIVITY
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetics
man26B gene
Man26B protein
mannanase
MANNANS
Mannosidases - biosynthesis
Mannosidases - genetics
Mannosidases - immunology
Mannosidases - metabolism
Mission oriented research
Molecular Sequence Data
NUCLEOTIDE SEQUENCE
plasmid pKS305
Protein Biosynthesis
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Summary: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 N-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 F1 cellulosome.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.65.548