Enhanced secretion of a methyl parathionhydrolase in Pichia pastoris using a combinationalstrategy

Microbial Cell Factories

Research

Ningfeng Wu¹  Lu Huang¹  Xiaoyu Chu¹  Jian Tian¹  Hu Jiang¹  Ping Wang¹ 

[1] Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, 100081, Beijing, China;

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

DOI  :  10.1186/s12934-015-0315-4

 received in 2015-01-26, accepted in 2015-08-10,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundAlthough Pichia pastoris has beensuccessfully used to produce various recombinant heterologous proteins, theefficiency varies. In this study, we used methyl parathion hydrolase (MPH) fromOchrobactrum sp. M231 as an example tostudy the effect of protein amino acid sequence on secretion from P. pastoris.ResultsThe results indicated that the protein N-terminal sequence, theendoplasmic reticulum (ER) retention signal (KKXX) at the protein C-terminus,and the acidic stability of the protein could affect its secretion fromP. pastoris. Mutations designed based onthese sequence features markedly improved secretion from P. pastoris. In addition, we found that the secretion propertiesof 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 greatlyimproved secretion and the secreted MPH activity of CHBD-DQR was enhanced up to195-fold compared with wild-type MPH without loss of catalyticefficiency.ConclusionsThese results demonstrate that the secretion of heterologousproteins from P. pastoris could be improvedby combining changes in multiple protein sequence features.

【 授权许可】

CC BY   
© Wang et al. 2015