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Overexpression of the regulatory subunit of protein kinase A increases heterologous protein expression in Pichia pastoris

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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 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.

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Supporting information

Supplementary Table 1—Primer sequences used in this study.

Supplementary Table 2—Selected translation-related factors of P. pastoris GS115.

Supplementary Table 3—Fold change (log2) in the mRNA expression of PKA-targeted genes.

Funding

This study was financially supported by the National Natural Science Foundation of China (Grant No. 31470159).

Author information

Authors and Affiliations

  1. Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China

    Xihao Liao, Wenyang Lin, Nanzhu Chen, Lu Li, Dafu Huang, Bingxu Zhong, Liang Chen, Shuli Liang & Ying Lin

  2. Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, 510006, Guangzhou, China

    Xihao Liao, Wenyang Lin, Nanzhu Chen, Lu Li, Dafu Huang, Bingxu Zhong, Liang Chen, Shuli Liang & Ying Lin

Authors
  1. Xihao Liao
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  2. Wenyang Lin
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  3. Nanzhu Chen
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  4. Lu Li
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  5. Dafu Huang
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  6. Bingxu Zhong
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  7. Liang Chen
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  8. Shuli Liang
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  9. Ying Lin
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Corresponding author

Correspondence to Ying Lin.

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The authors declare that they have no conflicts of interest.

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Liao, X., Lin, W., Chen, N. et al. Overexpression of the regulatory subunit of protein kinase A increases heterologous protein expression in Pichia pastoris. Biotechnol Lett 42, 2685–2692 (2020). https://doi.org/10.1007/s10529-020-02977-z

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  • DOI: https://doi.org/10.1007/s10529-020-02977-z

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