Construction and testing of Yarrowia lipolytica recombinant protein expression chassis cells based on the high-throughput screening and secretome

Microbial Cell Factories

Research

Ge Zhang¹  Qi Liu¹  Zongjie Dai¹  Siqian Yu²  Yingping Zhuang²  Jianye Xia³ 

[1] Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China;National Center of Technology Innovation for Synthetic Biology, 300308, Tianjin, China;State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, China;State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, China;Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 300308, Tianjin, China;National Center of Technology Innovation for Synthetic Biology, 300308, Tianjin, China;

关键词: Yarrowia lipolytica;    High-throughput screening;    Secretome;    Chassis cell;   

DOI  :  10.1186/s12934-023-02196-x

 received in 2023-06-04, accepted in 2023-09-05,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundIn the recombinant protein market with broad economic value, the rapid development of synthetic biology has made it necessary to construct an efficient exocrine expression system for the different heterologous proteins. Yarrowia lipolytica possesses unique advantages in nascent protein transport and glycosylation modification, so it can serve as a potential protein expression platform. Although the Po1 series derived from W29 is often used for the expression of the various heterologous proteins, the ability of W29 to secrete proteins has not been verified and the Po1 series has been found to be not convenient for further gene editing.ResultsA total of 246 Y. lipolytica strains were evaluated for their secretory capacity through performing high-throughput screening in 48-well plate. Thereafter, following two rounds of shake flask re-screening, a high-secreting protein starting strain DBVPG 5851 was obtained. Subsequently, combined with the extracellular protein types and relative abundance information provided by the secretome of the starting strain, available chassis cell for heterologous protein expression were preliminarily constructed, and it was observed that the most potential signal peptide was derived from YALI0D20680g.ConclusionsThis study offers a novel perspective on the diversification of Y. lipolytica host cells for the heterologous protein expression and provides significant basis for expanding the selection space of signal peptide tools in the future research.

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

【 图 表 】

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