Development of novel surface display platforms for anchoring heterologous proteins in Saccharomyces cerevisiae

Microbial Cell Factories

Xiaoyu Yang¹  Jin Hou¹  Yu Shen¹  Meihui Song¹  Hongting Tang²  Xiaoming Bao³ 

[1] 0000 0004 1761 1174, grid.27255.37, State Key Laboratory of Microbial Technology, Shandong University, 266237, Qingdao, People’s Republic of China;0000 0004 1761 1174, grid.27255.37, State Key Laboratory of Microbial Technology, Shandong University, 266237, Qingdao, People’s Republic of China;0000000119573309, grid.9227.e, Center for Synthetic Biochemistry, Chinese Academy of Sciences, Shenzhen Institutes for Advanced Technologies, 518055, Shenzhen, People’s Republic of China;0000 0004 1761 1174, grid.27255.37, State Key Laboratory of Microbial Technology, Shandong University, 266237, Qingdao, People’s Republic of China;Shandong Provincial Key Laboratory of Microbial Engineering, Qi Lu University of Technology, 250353, Jinan, People’s Republic of China;

关键词: Yeast surface display;    Aga1;    Dan4;    Sed1;    a-Agglutinin;    Glycosylphosphatidylinositol (GPI);   

DOI  :  10.1186/s12934-019-1133-x

来源: publisher

【 摘 要 】

BackgroundCell surface display of recombinant proteins has become a powerful tool for biotechnology and biomedical applications. As a model eukaryotic microorganism, Saccharomyces cerevisiae is an ideal candidate for surface display of heterologous proteins. However, the frequently used commercial yeast surface display system, the a-agglutinin anchor system, often results in low display efficiency.ResultsWe initially reconstructed the a-agglutinin system by replacing two anchor proteins with one anchor protein. By directly fusing the target protein to the N-terminus of Aga1p and inserting a flexible linker, the display efficiency almost doubled, and the activity of reporter protein α-galactosidase increased by 39%. We also developed new surface display systems. Six glycosylphosphatidylinositol (GPI) anchored cell wall proteins were selected to construct the display systems. Among them, Dan4p and Sed1p showed higher display efficiency than the a-agglutinin anchor system. Linkers were also inserted to eliminate the effects of GPI fusion on the activity of the target protein. We further used the newly developed Aga1p, Dan4p systems and Sed1p system to display exoglucanase and a relatively large protein β-glucosidase, and found that Aga1p and Dan4p were more suitable for immobilizing large proteins.ConclusionOur study developed novel efficient yeast surface display systems, that will be attractive tools for biotechnological and biomedical applications

【 授权许可】

CC BY