Enhancing the expression of recombinant kappa-carrageenase in Pichia pastoris using dual promoters, co-expressing chaperones and transcription factors

文献类型： 外文期刊

第一作者： Yu, Yuan

作者： Yu, Yuan;Yu, Yuan;Liu, Zhemin;Chen, Meng;Yang, Min;Li, Li;Mou, Haijin

作者机构：

关键词： kappa-carrageenase; Pichia pastoris expression system; GAP promoter; chaperones; transcription factors

期刊名称：BIOCATALYSIS AND BIOTRANSFORMATION （ 影响因子：2.181； 五年影响因子：2.104 ）

ISSN： 1024-2422

年卷期：

页码：

收录情况： SCI

摘要： In this study, with the aid of a constitutive promoter, and the co-expression of chaperone and transcription factor (TF) genes, the expression and enzymatic activity of recombinant kappa-carrageenase in Pichia pastoris containing truncated kappa-carrageenase gene cgkZ Delta Pst (GS115/pPIC9K-cgkZ Delta Pst) was enhanced. The recombinant P. pastoris strain containing constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter enabled the expression of recombinant kappa-carrageenase without methanol induction, the enzymatic activity was 2.73 U/mL after 96 h of shake flask fermentation at 22 degrees C. The enzymatic activity increased to 7.96 U/mL under methanol induction during P. pastoris growth, showing a 1.4-fold increase compared to that of the control group. With the co-expression of a series of chaperone genes and TFs that could promote protein folding, prevent protein aggregation, and counteract oxidative stress, the expression level of cgkZ Delta Pst showed a 1.29- to 1.93-fold increase from that in the control group. The enzymatic activity of the recombinant kappa-carrageenase increased to 7.07-7.70 U/mL. The use of the inducible P-AOX1 in combination with the constitutive P-GAP can further improve the productivity of recombinant kappa-carrageenase. The rational selection of molecular chaperones and TFs can also promote recombinant kappa-carrageenase secretion in P. pastoris. This work can be useful for the heterologous expression of other marine-origin glycoside hydrolases in P. pastoris.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序