期刊
GENE
卷 598, 期 -, 页码 50-62出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.gene.2016.10.040
关键词
Pichia pastoris; Recombinant expression; Secretion leader; Alpha mating factor; Circular dichroism; Protein modeling; Knob socket
资金
- NIH-AREA [GM65882-03]
- intramural Pacific Fund grant
- Pacific Department of Biological Sciences
The methylotrophic yeast Pichia pastoris has been used extensively for expressing recombinant proteins because it combines the ease of genetic manipulation, the ability to provide complex posttranslational modifications and the capacity for efficient protein secretion. The most successful and commonly used secretion signal leader in Pichia pastoris has been the alpha mating factor (MAT alpha) prepro secretion signal. However, limitations exist as some proteins cannot be secreted efficiently, leading to strategies to enhance secretion efficiency by modifying the secretion signal leader. Based on a Jpred secondary structure prediction and knob-socket modeling of tertiary structure, numerous deletions and duplications of the MAT alpha prepro leader were engineered to evaluate the correlation between predicted secondary structure and the secretion level of the reporters horseradish peroxidase (HRP) and Candida antarctica lipase B. In addition, circular dichroism analyses were completed for the wild type and several mutant pro-peptides to evaluate actual differences in secondary structure. The results lead to a new model of MAT alpha pro-peptide signal leader, which suggests that the N and C-termini of MAT alpha pro-peptide need to be presented in a specific orientation for proper interaction with the cellular secretion machinery and for efficient protein secretion. (C) 2016 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据