Abstract
An alkaline β-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°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 β-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.
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Acknowledgements
We thank Prof. W.D. Grant for his kind advice. This work was supported by the Chinese Academy of Sciences, by the Ministry of Science and Technology of China, and by the National Basic Research Program of China (no. 2003CB716001).
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Ma, Y., Xue, Y., Dou, Y. et al. Characterization and gene cloning of a novel β-mannanase from alkaliphilic Bacillus sp. N16-5. Extremophiles 8, 447–454 (2004). https://doi.org/10.1007/s00792-004-0405-4
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DOI: https://doi.org/10.1007/s00792-004-0405-4