2006•Zhongguo shengwu gongcheng zazhiRequires access
Secreted Expression of M annanase Gene in Pichia pastoris and Anylysis of Enzymic Properties
Ming Yue
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Abstract
A PCR method was used to amplify the sequence encoding the mature peptide ofβ-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream ofα-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.