标题：Secretory pathway engineering enhances secretion of cellobiohydrolase I from Trichoderma reesei in Saccharomyces cerevisiae
作者：Lili Xu;Yu Shen;Jin Hou
作者机构：[Xu, L] The State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China;[ Shen, Y] The State Key Laboratory of Microbial Te 更多
通讯作者地址：[Bao, XM]Shandong Univ, State Key Lab Microbial Technol, Jinan 250100, Peoples R China.
来源：Journal of Bioscience and Bioengineering
关键词：Cel7A;Protein secretion;PMR1;Consolidated bioprocessing;Cellulosic ethanol;Budding yeast
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 5. cerevisiae. The effects of these modifications on the secretion of cellobiohydrolase I (7r-Cel7A) with its native signal peptide from Trichoderma reesei were investigated. The results showed that overexpression of the protein disulfide isomerase Sc-PDIl and the plasma membrane targeting soluble N-ethylmaleimide-sensitive factor attachment protein receptor Sc-SSOl, and disruption of the sorting receptor Sc-VPS10 and a Ca~(2+)/Mn~(2+) ATPase Sc-PMRl, improved respectively the extracellular Tr-Cel7A activities. Among them, disruption of Sc-PMRl 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 (ypslOAIpmrlAISSOllPDn/cel17AF) using p-nitrophenyl-P-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 7r-Cel7A secretion in S. cerevisiae.