Abstract
β-Mannanase (EC 3.2.1.78) is a key enzyme to hydrolyze the β-mannosidic linkages in mannan and heteromannan. The expression of a wild type β-mannanase (manWT) of Aspergillus sulphureus in Pichia pastoris is not high enough for its application in feed supplement. To earn a high expression level, the manWT gene was firstly optimized to manM according to the code bias of P. pastoris, which was then inserted into pPICzαA and transformed into P. pastoris strain X-33. In the induction by methanol, β-mannanase was expressed in high level with 32% increase in comparison with the manWT gene expressed in P. pastoris in shaken flask. In a 10-L fermenter, the manM was expressed in 9-fold higher level than that in shaken flask, which yielded the enzyme activity of 1100 U/mL. This is the first study on codon bias effect on the β-mannanase gene expression level, which helps to achieve high β-mannanase yield and enzymatic activity in P. pastoris.
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Abbreviations
- manM:
-
modified β-mannanase gene
- manWT:
-
wild type β-mannanase gene
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Chen, X., Qiao, J., Yu, H. et al. Overexpression of an optimized Aspergillus sulphureus β-mannanase gene in Pichia pastoris . Biologia 64, 235–238 (2009). https://doi.org/10.2478/s11756-009-0043-5
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DOI: https://doi.org/10.2478/s11756-009-0043-5