Improvement of ATP regeneration efficiency and operation stability in porcine interferon-α production by Pichia pastoris under lower induction temperature

Minjie Gao; Shijuan Dong; Ruisong Yu; Jianrong Wu; Zhiyong Zheng; Zhongping Shi; Xiaobei Zhan

Korean Journal of Chemical Engineering, Vol.28, No.6, 1412-1419, June, 2011

DOI10.1007/s11814-010-0527-6 Export Citation

Improvement of ATP regeneration efficiency and operation stability in porcine interferon-α production by Pichia pastoris under lower induction temperature

Minjie Gao, Shijuan Dong¹, Ruisong Yu¹, Jianrong Wu, Zhiyong Zheng, Zhongping Shi^†, and Xiaobei Zhan

Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China
¹Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Science, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Shanghai 201106, China

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The performance of traditional heterologous protein production by Pichia pastoris with methanol induction at 30 ℃ is poor, characterized by low ATP regeneration rate and weak operation stability. A low temperature induction strategy at 20 ℃ was thus adopted for efficient porcine interferon-α production in a 10 L fermentor. With the strategy, maximal methanol tolerance level could reach about 40 g/L to effectively deal with methanol concentration variations, so that the complicated on-line methanol measurement system could be eliminated. Moreover, metabolic analysis based on multiple state-variables measurements indicated that pIFN-α antiviral activity enhancement profited from the formation of an efficient ATP regeneration system at 20 ℃induction. Compared to the induction strategy at 30 ℃, the proposed strategy increased the ATP regeneration rate by 49-66%, the maximal pIFN-α antiviral activity was enhanced about 20-fold and reached a higher level of 1.5×106 IU/mL.

Keywords:Fed-batch Culture;Fermentation;Pichia pastoris;Porcine Interferon-α;Protein Expression

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[2] Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM, Yeast., 22, 249 (2005)

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[4] Chinsangaram J, Moraes MP, Koster M, Grubman MJ, J. Virol., 77, 1621 (2003)

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[13] Wang Y, Wang ZH, Xu QL, Du GC, Hua ZZ, Liu LM, Li JH, Chen J, Process. Biochem., 44, 949 (2009)

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[22] Suye S, Ogawa A, Yokoyama S, Obayashi A, Agric. Biol. Chem., 54, 1297 (1990)

[23] Duan SB, Shi ZP, Feng HJ, Duan ZY, Mao ZG, Biochem. Eng. J., 30, 88 (2006)

[24] Jin H, Zheng ZY, Gao MJ, Duan ZY, Shi ZP, Wang ZX, Jin J, Biochem. Eng. J., 37, 26 (2007)

[25] Charoenrat T, Ketudat-Cairns M, Stendahl-Andersen H, Jahic M, Enfors SO, Bioprocess Biosyst. Eng., 27, 399 (2005)

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