Engineering protein folding and translocation improves heterologous protein secretion in Saccharomyces cerevisiae

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Abstract
Saccharomyces cerevisiae is widely used as a producer of heterologous proteins of medical and industrial interest. Numerous efforts have been made to overcome bottlenecks in protein expression and secretion. However, the effect of engineering protein translocation to heterologous protein secretion has not been studied extensively in S. cerevisiae. In this work, we confirmed that heterologous protein expression in S. cerevisiae induced the unfolded protein response (UPR). To enhance protein folding capacity, the endoplasmic reticulum (ER) chaperone protein BiP and the disulfide isomerase Pdi1p were each over‐expressed, and the secretion of three heterologous proteins, β‐glucosidase, endoglucanase, and α‐amylase, was improved. The impact of engineering key translocation components was also studied. The over‐expression of co‐translational translocation components Srp14p and Srp54p enhanced the secretion of three heterologous proteins (β‐glucosidase, endoglucanase, and α‐amylase), but over‐expressing the cytosolic chaperone Ssa1p (involved in post‐translational translocation) only enhanced the secretion of β‐glucosidase. By engineering both co‐translational translocation and protein folding, we obtained strains with β‐glucosidase, endoglucanase, and α‐amylase activities increased by 72%, 60%, and 103% compared to the controls. Our results show that protein translocation may be a limiting factor for heterologous protein production. Biotechnol. Bioeng. 2015;112: 1872–1882.
Funding Information
  • National Key Technology R&D Program of China (2014BAD02B07)
  • National High Technology Research and Development Program of China (2012AA022106 2014AA021903)
  • National Natural Science Foundation of China (31300037 31470219 J1103515)
  • Project of National Energy Administration of China (NY20130402)
  • Promotive research fund for excellent young, Middle-aged scientists of Shandong Province (BS2013SW020)
  • State Key Laboratory for Microbial Technology (M2013-08)
  • Shandong Key Laboratory of Straw Biorefinement

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