Overexpression of the regulatory subunit of protein kinase A increases heterologous protein expression in Pichia pastori
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ORIGINAL RESEARCH PAPER
Overexpression of the regulatory subunit of protein kinase A increases heterologous protein expression in Pichia pastoris Xihao Liao . Wenyang Lin . Nanzhu Chen . Lu Li . Dafu Huang . Bingxu Zhong . Liang Chen . Shuli Liang . Ying Lin
Received: 9 February 2020 / Accepted: 27 July 2020 Ó Springer Nature B.V. 2020
Abstract Objective Translational regulation plays an important role in protein synthesis. Our goal was to screen translation-related factors to improve heterologous protein expression in Pichia pastoris. Results Twenty-eight translation-related factors were overexpressed in P. pastoris GS115 expressing enhanced green fluorescent protein (eGFP). The results showed that overexpression of Bcy1, the regulatory subunit of protein kinase A (PKA), significantly increased both eGFP expression and cell biomass by 20% under methanol induction for 120 h. Additionally, overexpression of Bcy1 elevated the growth rate by 18% and increased production of
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02977-z) contains supplementary material, which is available to authorized users. X. Liao W. Lin N. Chen L. Li D. Huang B. Zhong L. Chen S. Liang Y. Lin (&) Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006 Guangzhou, China e-mail: [email protected] X. Liao W. Lin N. Chen L. Li D. Huang B. Zhong L. Chen S. Liang Y. Lin Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006 Guangzhou, China
the industrial enzyme Phytase (Phy) by 26%. Transcriptome analysis indicated that the overall expression of ribosomal protein genes was significantly downregulated and that postdiauxic shift genes and stress response element genes were upregulated. Conclusions Bcy1 regulates ribosome protein genes, postdiauxic shift genes and stress response element genes, leading to improved cell growth and heterologous protein expression. This study provides a convenient and universal factor for heterologous protein production. Keywords Heterologous protein Pichia pastoris Translation-related factor Bcy1
Introduction Pichia pastoris is a universal and efficient heterologous protein expression system widely used in the production of various pharmaceutical proteins and industrial enzymes (Juturu and Wu 2018). In addition to the metabolic engineering of P. pastoris, current strategies for improving heterologous protein synthesis mainly focus on gene transcription, protein folding and secretion (Yang and Zhang 2018). However, with the recognition that mRNA and protein abundance in various species is poorly correlated, with R2 ranging from * 0.01 to 0.50 (Maier et al. 2009), translation is a key node in the flow of genetic information that
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Biotechnol Lett
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