SCI和EI收录∣中国化工学会会刊

Chinese Journal of Chemical Engineering ›› 2013, Vol. 21 ›› Issue (2): 216-226.DOI: 10.1016/S1004-9541(13)60461-9

• 综述 • 上一篇    

Process Control and Optimization for Heterologous Protein Production by Methylotrophic Pichia pastoris

高敏杰, 史仲平   

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
  • 收稿日期:2012-09-18 修回日期:2012-11-05 出版日期:2013-02-28 发布日期:2013-03-13
  • 通讯作者: SHI Zhongping
  • 基金资助:

    Supported by the Key Agricultral Technology Program of Shanghai Science & Technology Committee (073919108) and Major State Basic Research Development Program of China (2007CB714303).

Process Control and Optimization for Heterologous Protein Production by Methylotrophic Pichia pastoris

GAO Minjie, SHI Zhongping   

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
  • Received:2012-09-18 Revised:2012-11-05 Online:2013-02-28 Published:2013-03-13

摘要: The methylotrophic yeast Pichia pastoris is a highly successful system for production of a variety of heterologous proteins due to its unique features/abilities for effective protein expression, and tremendous efforts have been made to increase heterologous protein productivity by P. pastoris in recent years. When new engineered yeast strains are constructed and are ready to use for industrial protein production, process control and optimization techniques should be applied to improve the fermentation performance in the following aspects: (1) increase recombinant cell concentrations in fermentor to high density during growth phase; (2) effectively induce heterologous proteins by enhancing/stabilizing titers or concentrations of the proteins during induction phase; (3) decrease operation costs by relieving the working loads of heat-exchange and oxygen supply. This article reviews and discusses the key and commonly used techniques in heterologous protein production by P. pastoris, with the focus on optimizations of fermentation media and basic operation conditions, development of optimal glycerol feeding strategies for achieving high density cultivation of P. pastoris and effective heterologous protein induction methods by regulating specific growth rate, methanol concentration, temperatures, mixture ratio of multi-carbon substrates, etc. Metabolic analysis for recombinant protein production by P. pastoris is also introduced to interpret the mechanism of sub-optimal heterologous protein production and to explore further optimal expression methods.

关键词: fermentation, heterologous protein, metabolic analysis, Pichia pastoris, process control and optimization

Abstract: The methylotrophic yeast Pichia pastoris is a highly successful system for production of a variety of heterologous proteins due to its unique features/abilities for effective protein expression, and tremendous efforts have been made to increase heterologous protein productivity by P. pastoris in recent years. When new engineered yeast strains are constructed and are ready to use for industrial protein production, process control and optimization techniques should be applied to improve the fermentation performance in the following aspects: (1) increase recombinant cell concentrations in fermentor to high density during growth phase; (2) effectively induce heterologous proteins by enhancing/stabilizing titers or concentrations of the proteins during induction phase; (3) decrease operation costs by relieving the working loads of heat-exchange and oxygen supply. This article reviews and discusses the key and commonly used techniques in heterologous protein production by P. pastoris, with the focus on optimizations of fermentation media and basic operation conditions, development of optimal glycerol feeding strategies for achieving high density cultivation of P. pastoris and effective heterologous protein induction methods by regulating specific growth rate, methanol concentration, temperatures, mixture ratio of multi-carbon substrates, etc. Metabolic analysis for recombinant protein production by P. pastoris is also introduced to interpret the mechanism of sub-optimal heterologous protein production and to explore further optimal expression methods.

Key words: fermentation, heterologous protein, metabolic analysis, Pichia pastoris, process control and optimization