Secretory pathway engineering enhances secretion of cellobiohydrolase I from Trichoderma reesei in Saccharomyces cerevisiae

被引:38
作者
Xu, Lili [1 ]
Shen, Yu [1 ]
Hou, Jin [1 ]
Peng, Bingyin [1 ]
Tang, Hongting [1 ]
Bao, Xiaoming [1 ]
机构
[1] Shandong Univ, State Key Lab Microbial Technol, Jinan 250100, Peoples R China
基金
中国国家自然科学基金;
关键词
Cel7A; Protein secretion; PMR1; Consolidated bioprocessing; Cellulosic ethanol; Budding yeast; ENDOPLASMIC-RETICULUM; HETEROLOGOUS PROTEINS; EFFICIENT SECRETION; YEAST; EXPRESSION; GLYCOSYLATION; FERMENTATION; CELLULOSE; ASSAY; PMR1;
D O I
10.1016/j.jbiosc.2013.06.017
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Improving the cellulase secretion is beneficial for Saccharomyces cerevisiae used in consolidated bioprocessing (CBP) of cellulosic ethanol. In this study, protein secretory pathway, including protein folding, disulfide bond formation, and protein trafficking and sorting, was modified in S. cerevisiae. The effects of these modifications on the secretion of cellobiohydrolase I (Tr-Cel7A) with its native signal peptide from Trichoderma reesei were investigated. The results showed that overexpression of the protein disulfide isomerase Sc-PDI1 and the plasma membrane targeting soluble N-ethylmaleimide-sensitive factor attachment protein receptor Sc-SSO1, and disruption of the sorting receptor Sc-VPS10 and a Ca2+/Mn2+ ATPase Sc-PMR1, improved respectively the extracellular Tr-Cel7A activities. Among them, disruption of Sc-PMR1 showed better improvement of 162% in the extracellular activity and decreased the glycosylation of Tr-Cel7A. Multiple modifications generally resulted in higher activities. The extracellular activities of the quadruple-modified strain (vps10 Delta/pmr1 Delta/SSO1/PDI1/cel7AF) using p-nitrophenyl-beta-D-cellobioside (pNPC) and phosphoric acid swollen cellulose (PASC) as the substrates, respectively, were 3.9-fold and 1.3-fold higher than that of the reference strain cel7AF. The results indicated that engineering of the protein secretory pathway is an effective approach to improve the Tr-Cel7A secretion in S. cerevisiae. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.
引用
收藏
页码:45 / 52
页数:8
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