Characterization and gene cloning of a novel β-mannanase from alkaliphilic Bacillus sp. N16-5

An alkaline beta-mannanase was purified to homogeneity from a culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme had optimum activity at pH 9.5 and 70 degrees C. It was composed of a single polypeptide chain with a molecular weight of 55 kDa deduced from SDS-PAGE, and its isoelectric point...

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Bibliographic Details
Published in:Extremophiles : life under extreme conditions 2004-12, Vol.8 (6), p.447-454
Main Authors: Ma, Yanhe, Xue, Yanfen, Dou, Yuetan, Xu, Zhenghong, Tao, Wenyi, Zhou, Peijin
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacillus
Bacillus - classification
Bacillus - enzymology
Bacillus - genetics
Base Sequence
beta-Mannosidase - chemistry
beta-Mannosidase - genetics
beta-Mannosidase - metabolism
Biological and medical sciences
Catalytic Domain - genetics
Cloning, Molecular
Conserved Sequence
DNA, Bacterial - genetics
Enzyme Stability
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Hydrogen-Ion Concentration
Kinetics
Metals
Microbiology
Molecular Sequence Data
Molecular Weight
Open Reading Frames
Sequence Homology, Amino Acid
Space life sciences
Substrate Specificity
Temperature
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Summary:An alkaline beta-mannanase was purified to homogeneity from a culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme had optimum activity at pH 9.5 and 70 degrees C. It was composed of a single polypeptide chain with a molecular weight of 55 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 4.3. The enzyme efficiently hydrolyzed galactomannan and glucomannan, producing a series of oligosaccharides and monosaccharides. The beta-mannanase gene (manA) contained an open reading frame (ORF) of 1,479 bp, encoding a 32-amino acids signal peptide, and a mature protein of 461 amino acids, with a calculated molecular mass of 50,743 Da. Strain N16-5 ManA, deduced from the manA ORF, exhibited relatively high amino acid similarity to the members of the glycosyl hydrolase family 5. The eight conserved active-site amino acids in family 5 glycosyl hydrolase were found in the deduced amino acid sequence of strain N16-5 ManA.
ISSN:1431-0651
1433-4909
DOI:10.1007/s00792-004-0405-4