文献类型：期刊文献

英文题名：Activation of the Unfolded Protein Response via Co-expression of the HAC1(i) Gene Enhances Expression of Recombinant Elastase in Pichia pastoris

作者：Han, Minghai Wang, Weixian Zhou, Jianli Gong, Xun Xu, Cunbin Li, Yinfeng Li, Qiang

第一作者：韩铭海

通信作者：Han, MH[1]|[144402eaf094eaf27b6d2]韩铭海;

机构：[1]Guizhou Inst Technol, Coll Food & Pharmaceut Engn, Guiyan 550000, Peoples R China;[2]Guangdong Prov Key Lab Fermentat & Enzyme Engn, Guangzhou 510006, Peoples R China;[3]Jiangnan Univ, Sch Biotechnol, Wuxi 214122, Jiangsu, Peoples R China

第一机构：贵州理工学院食品药品制造工程学院

通信机构：corresponding author), Guizhou Inst Technol, Coll Food & Pharmaceut Engn, Guiyan 550000, Peoples R China.|贵州理工学院食品药品制造工程学院;贵州理工学院;

年份：2020

卷号：25

期号：2

起止页码：302-307

外文期刊名：BIOTECHNOLOGY AND BIOPROCESS ENGINEERING

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000526329200001)】；

基金：This study was funded by the National Natural Science Foundation of China (Grant No. 31760022) and by Guizhou Provincial Science and Technology Foundation (Grant No. [2019]1146).

语种：英文

外文关键词：unfolded protein response; Pichia pastoris; protein expression; HAC1

摘要：The effects of activation of the unfolded protein response (UPR) via co-expression of the HAC1(i) gene on the production of the recombinant Pseudomonas aeruginosa elastase (rPAE) in Pichia pastoris GS115 were evaluated in this study. The results showed that expression of the HAC1(i) gene significantly increased the level of Kar2p (a hallmark of UPR activation) in P. pastoris GS115, demonstrating activation of the UPR. This gene did not affect the growth of yeast in the buffered glycerol-complex medium but stimulated its growth in the buffered methanol-complex medium. Co-expression of the HAC1(i) gene enhanced the expression level of the heterogeneously N-glycosylated forms of rPAE, as the caseinolytic activity in the supernatant of the various glycoforms of rPAE expressed in P. pastoris GS115/HAC1 was increased 1.8-3.9-fold compared to that in the control strain P. pastoris GS115, respectively. The stimulating effects of co-expression of the gene on rPAE production were observed when 0.5, 1.0, and 2.0% methanol were added every 24 h, as the caseinolytic activity of supernatants of P. pastoris GS115/HAC1 expressing wild-type of rPAE was increased 3.3-, 1.9-, and 1.7-fold at the corresponding methanol concentration. Further, activation of UPR via co-expression of the HAC1(i) gene enhanced rPAE secretion in P. pastoris at 20, 24, and 28 degrees C, as the caseinolytic activity of supernatants of P. pastoris GS115/HAC1 expressing wild-type rPAE was increased 2.3-, 2.1-, and 2.8-fold over the tested temperatures.