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Improvement of specific growth rate of Pichia pastoris for effective porcine interferon-alpha production with an on-line model-based glycerol feeding strategy

文献类型： 外文期刊

作者： Gao, Min-Jie 1 ; Zheng, Zhi-Yong 1 ; Wu, Jian-Rong 1 ; Dong, Shi-Juan 2 ; Li, Zhen 2 ; Jin, Hu 1 ; Zhan, Xiao-Bei 1 ; Lin 1 ;

作者机构： 1.Jiangnan Univ, Minist Educ, Key Lab Ind Biotechnol, Wuxi 214122, Peoples R China

2.Shanghai Acad Agr Sci, Anim Husb & Vet Res Inst, Shanghai Municipal Key Lab Agri Genet & Breeding, Shanghai 201106, Peoples R China

关键词： DO-stat;Fed-batch culture;Pichia pastoris;Protein expression;Artificial neural network

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2012 年 93 卷 4 期

页码：

收录情况： SCI

摘要： Effective expression of porcine interferon-alpha (pIFN-alpha) with recombinant Pichia pastoris was conducted in a bench-scale fermentor. The influence of the glycerol feeding strategy on the specific growth rate and protein production was investigated. The traditional DO-stat feeding strategy led to very low cell growth rate resulting in low dry cell weight (DCW) of about 90 g/L during the subsequent induction phase. The previously reported Artificial Neural Network Pattern Recognition (ANNPR) model-based glycerol feeding strategy improved the cell density to 120 g DCW/L, while the specific growth rate decreased from 0.15 to 0.18 to 0.03-0.08 h(-1) during the last 10 h of the glycerol feeding stage leading to a variation of the porcine interferon-alpha production, as the glycerol feeding scheme had a significant effect on the induction phase. This problem was resolved by an improved ANNPR model-based feeding strategy to maintain the specific growth rate above 0.11 h(-1). With this feeding strategy, the pIFN-alpha concentration reached a level of 1.43 g/L, more than 1.5-fold higher than that obtained with the previously adopted feeding strategy. Our results showed that increasing the specific growth rate favored the target protein production and the glycerol feeding methods directly influenced the induction stage. Consequently, higher cell density and specific growth rate as well as effective porcine interferon-alpha production have been achieved by our novel glycerol feeding strategy.

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[1]Improvement of ATP regeneration efficiency and operation stability in porcine interferon-alpha production by Pichia pastoris under lower induction temperature. Gao, Minjie,Wu, Jianrong,Zheng, Zhiyong,Shi, Zhongping,Zhan, Xiaobei,Dong, Shijuan,Yu, Ruisong. 2011

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[4]Enhancing pIFN-alpha Production and Process Stability in Fed-Batch Culture of Pichia pastoris by Controlling the Methanol Concentration and Monitoring the Responses of OUR/DO Levels. Gao, Min-Jie,Zhan, Xiao-Bei,Zheng, Zhi-Yong,Wu, Jian-Rong,Shi, Zhong-Ping,Lin, Chi-Chung,Dong, Shi-Juan,Li, Zhen. 2013

[5]High expression of a heat-stable phytase in Pichia pastoris. Peng, RH,Xiong, AS,Li, X,Fan, HQ,Yao, QH,Guo, MJ,Zhang, SL. 2002

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[19]Improvement of Porcine Interferon-alpha Production by Recombinant Pichia pastoris via Induction at Low Methanol Concentration and Low Temperature. Jin, Hu,Liu, Guoqiang,Dai, Keke,Wang, Huihui,Shi, Zhongping,Li, Zhen. 2011

[20]Enhanced porcine interferon-alpha production by recombinant Pichia pastoris with a combinational control strategy of low induction temperature and high dissolved oxygen concentration. Jin, Hu,Liu, Guoqiang,Duan, Zuoying,Shi, Zhongping,Ye, Xiaofei,Li, Zhen. 2010

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