Alkaline mannanase from a novel species of alkaliphilic Bacillus

A novel species of alkaliphilic Bacillus (JAMB-602), isolated from a soil sample, was found to exoproduce an alkaline mannanase. The mannanase gene was cloned by the shotgun method and sequenced. Its open reading frame encodes a protein of 490 amino acids with a calculated molecular mass of 53,763 D...

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Bibliographic Details
Published inJournal of Applied Glycoscience Vol. 51; no. 3; pp. 229 - 236
Main Authors Takeda, N. (Toyo Univ., Tokyo (Japan)), Hirasawa, K, Uchimura, K, Nogi, Y, Hatada, Y, Akita, M, Usami, R, Yoshida, Y, Grant, W.D, Ito, S, Horikoshi, K
Format Journal Article
LanguageEnglish
Japanese
Published The Japanese Society of Applied Glycoscience 2004
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Summary:A novel species of alkaliphilic Bacillus (JAMB-602), isolated from a soil sample, was found to exoproduce an alkaline mannanase. The mannanase gene was cloned by the shotgun method and sequenced. Its open reading frame encodes a protein of 490 amino acids with a calculated molecular mass of 53,763 Da. The deduced amino acid sequence showed similarity to those of known mannanases in glycoside hydrolase family 5, with only 32-58% identity. The recombinant mature enzyme with a molecular mass of 50 kDa was hyperexoproduced with Bacillus subtilis as the host, corresponding to a level of approximately 2 g/L. The homogeneously purified enzyme had a molecular mass of 50 kDa, an optimal temperature of 65 deg C, and an optimal pH of around 9. It was thermostable with a half-life of 2 h at 55 deg C. N -Bromosuccinimide abolished the enzymatic activity. The pattern of mannan hydrolysis showed that the enzyme is an endo-type mannanase.
Bibliography:F60
F30
2005002116
ISSN:1344-7882
1880-7291
DOI:10.5458/jag.51.229