Enhanced secretion of a methyl parathion hydrolase in Pichia pastoris using a combinational strategy

文献类型： 外文期刊

第一作者： Wang, Ping

作者： Wang, Ping;Huang, Lu;Jiang, Hu;Tian, Jian;Chu, Xiaoyu;Wu, Ningfeng

作者机构：

关键词： Methyl parathion hydrolase;Cytochromes heme binding domain;KKXX retrieval signal;Acid stability;Secretory expression;Pichia pastoris

期刊名称：MICROBIAL CELL FACTORIES （ 影响因子：5.328； 五年影响因子：5.588 ）

ISSN： 1475-2859

年卷期： 2015 年 14 卷

页码：

收录情况： SCI

摘要： Background: Although Pichia pastoris has been successfully used to produce various recombinant heterologous proteins, the efficiency varies. In this study, we used methyl parathion hydrolase (MPH) from Ochrobactrum sp. M231 as an example to study the effect of protein amino acid sequence on secretion from P. pastoris. Results: The results indicated that the protein N-terminal sequence, the endoplasmic reticulum (ER) retention signal (KKXX) at the protein C-terminus, and the acidic stability of the protein could affect its secretion from P. pastoris. Mutations designed based on these sequence features markedly improved secretion from P. pastoris. In addition, we found that the secretion properties of a protein can be cumulative when all of the above strategies are combined. The final mutant (CHBD-DQR) designed by combining all of the strategies greatly improved secretion and the secreted MPH activity of CHBD-DQR was enhanced up to 195-fold compared with wild-type MPH without loss of catalytic efficiency. Conclusions: These results demonstrate that the secretion of heterologous proteins from P. pastoris could be improved by combining changes in multiple protein sequence features.

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