2. Academic Validation
  3. Enhancing co-translational folding of heterologous protein by deleting non-essential ribosomal proteins in Pichia pastoris

Enhancing co-translational folding of heterologous protein by deleting non-essential ribosomal proteins in Pichia pastoris

  • Biotechnol Biofuels. 2019 Feb 21;12:38. doi: 10.1186/s13068-019-1377-z.
Xihao Liao 1 2,  Jing Zhao 3,  Shuli Liang 1 2,  Jingjie Jin 3,  Cheng Li 1 2,  Ruiming Xiao 1 2,  Lu Li 1 2,  Meijin Guo 4,  Gong Zhang 3,  Ying Lin 1 2
Affiliations

Affiliations

  • 1 1Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006 China.
  • 2 2Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006 China.
  • 3 3Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou, 510632 China.
  • 4 4State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai Institute of Biomanufacturing Technology & Collaborative Innovation Center, Shanghai, 200237 China.
PMID: 30828383 DOI: 10.1186/s13068-019-1377-z
Abstract

Background: Translational regulation played an important role in the correct folding of heterologous proteins to form bioactive conformations during biogenesis. Translational pausing coordinates protein translation and co-translational folding. Decelerating translation elongation speed has been shown to improve the soluble protein yield when expressing heterologous proteins in industrial expression hosts. However, rational redesign of translational pausing via synonymous mutations may not be feasible in many cases. Our goal was to develop a general and convenient strategy to improve heterologous protein synthesis in Pichia pastoris without mutating the expressed genes.

Results: Here, a large-scale deletion library of ribosomal protein (RP) genes was constructed for heterologous protein expression in Pichia pastoris, and 59% (16/27) RP deletants have significantly increased heterologous protein yield. This is due to the delay of 60S subunit assembly by deleting non-essential ribosomal protein genes or 60S subunit processing factors, thus globally decreased the translation elongation speed and improved the co-translational folding, without perturbing the relative transcription level and translation initiation.

Conclusion: Global decrease in the translation elongation speed by RP deletion enhanced co-translational folding efficiency of nascent chains and decreased protein aggregates to improve heterologous protein yield. A potential expression platform for efficient pharmaceutical proteins and industrial enzymes production was provided without synonymous mutation.

Keywords

Co-translational folding; Heterologous protein; Pichia pastoris; Ribosomal protein; Translation.

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