文献类型：期刊文献

英文题名：Screening endogenous signal peptides and protein folding factors to promote the secretory expression of heterologous proteins in Pichia pastoris

作者：Duan, Guangdong[1];Ding, Lumei[1];Wei, Dongsheng[1];Zhou, Hangcheng[1];Chu, Ju[1];Zhang, Siliang[1];Qian, Jiangchao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：306

起止页码：193

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20194307579293);WOS:【SCI-EXPANDED(收录号:WOS:000496193800024)】；

基金：We are grateful to National Basic Research Program (973 Program 2013CB733600) and the Fundamental Research Funds for the Central Universities (22221818014) for the financial supports to this research. We thank Dr. James M. Cregg for his generous gift of the plasmid pSAOH5.

语种：英文

外文关键词：Signal peptides; Folding factors; Pichia pastoris; Heterologous protein; Secretory expression

摘要：Secretory expression is most often desired but usually hampered by limitations of signal peptide processing and protein folding in the methylotrophic yeast Pichia pastoris. To alleviate such limitations, novel endogenous signal peptides (Dan4, Gas1, Msb2, and Fre2) and folding factors (Mpd1p, Pdi2p, and Sil1p) were predicted based on the reported P. pastoris secretome and genome. Their effects were investigated using three reporter proteins: yeast-enhanced green fluorescent protein (yEGFP), beta-galactosidase (Gal) and cephalosporin C acylase (SECA), in comparison with the commonly used Saccharomyces cerevisiae alpha-mating factor pre-pro leader sequence (alpha-MF) or folding factors (Pdi1p, BiP, and Hac1p). The newly identified signal sequences were superior over alpha-MF for production of heterologous proteins. The signal peptide Msb2 increased the specific extracellular production of all reporter proteins, ranging from 1.5- to 8.0-fold, and Dan4 enhanced all total protein production up to 172-fold. Co-expression of folding factors exhibited a protein-specific effect on cell growth, transcription and expression of different reporter genes. All of the novel folding factors enhanced total production of SECA, and Sil1p performed best in the extracellular SECA production, showing a 3.3-fold increase. These novel signal peptides and folding factors can be used for promoting secretion of heterologous proteins in P. pastoris.