Increased production of alkaline polygalacturonate lyase in the recombinant Pichia pastoris by controlling cell concentration during continuous culture

详细信息    查看全文

• 作者：Huilin Wang^a ; ^b ; Jianghua Li^c ; Long Liu^c ; Xiaoman Li^a ; ^b ; Dongxu Jia^c ; Guocheng Du^c ; ^d ; ^{gcdu@jiangnan.edu.cn} ; Jian Chen^c ; ^d ; ^{jchen@jiangnan.edu.cn} ; Jiangning Song^a ; ^b ; ^e ; ^{song_jn@tib.cas.cn}
• 关键词：Pichia pastoris ; Alkaline polygalacturonate lyase (PGL) ; Cell concentration ; Continuous culture ; Productivity
• 刊名：Bioresource Technology
• 出版年：2012
• 出版时间：November, 2012
• 年：2012
• 卷：124
• 期：Complete
• 页码：338-346
• 全文大小：528 K

文摘

Recombinant alkaline polygalacturonate lyase (PGL) production by recombinant Pichia pastoris GS115 was selected as a model to study as a continuous culture strategy for enhancing heterologous protein production based on controlling methanol feeding (CCCM culture) or on dual carbon source feeding (CCCD culture). Using the CCCM process with a dry cell weight of 75 g/L regulated by controlling methanol concentration in the induction media, the final PGL activity was 441.9 U/mL. The PGL productivity (Q_v) and the average specific enzyme production rate (Q_x) were 4.65 U mL^?1 h^?1 and 84.5 U g^?1 h^?1, an increase of 42.1 % and 191.2 % , respectively, over what was achieved with traditional fed-batch culture with high cell density. The control strategies also reduced proteolytic degradation by 84.1 % in the fermentation broth and increased cell viability by 12.2 % .