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

• Published in: Chinese journal of chemical engineering Vol. 21; no. 2; pp. 216 - 226
• Main Authors: GAO, Minjie, SHI, Zhongping
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.02.2013
• Subjects: fermentation; Heat exchangers; heterologous protein; High density; metabolic analysis; Optimization; Pichia pastoris; Process control; process control and optimization; Proteins; Recombinant; Yeast; 优化技术; 外源蛋白; 异源蛋白; 生产过程控制; 甲醇毕赤酵母; 细胞浓度; 重组蛋白; 高密度发酵; metabolic analysis; fermentation; process control and optimization; Pichia pastoris; heterologous protein
• ISSN: 1004-9541; 2210-321X
• DOI: 10.1016/S1004-9541(13)60461-9

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 tot 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.