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Methanol/sorbitol co-feeding induction enhanced porcine interferon-α production by P. pastoris associated with energy metabolism shift

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Abstract

The production of porcine interferon-α (pIFN-α) by Pichia pastoris was largely enhanced when adopting sorbitol/methanol co-feeding induction strategy at 30 °C in a 10-L fermentor. Analysis of energy regeneration pattern and carbon metabolism revealed that major energy metabolism energizing pIFN-α synthesis shifted from formaldehyde dissimilatory energy metabolism pathway to TCA cycle under the methanol/sorbitol co-feeding induction strategy. The sorbitol/methanol co-feeding induction strategy weakened formaldehyde dissimilatory pathway and repressed the accumulation of toxic metabolite-formaldehyde, reduced theoretical oxygen consumption rate and oxygen supply requirement, and increased energy/methanol utilization efficiency so that more methanol could be effectively used for pIFN-α synthesis. As a result, pIFN-α antiviral activity reached a highest level of 1.8 × 107 IU/mL which was about 10- to 200-folds of those obtained under pure methanol induction at 20 and 30 °C, respectively.

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Abbreviations

AOX:

Alcohol oxidase

CER:

CO2 evolution rate

DCW:

Dry cell weight

FDH:

Formate dehydrogenase

FLD:

Formaldehyde dehydrogenase

HCHO:

Formaldehyde

HCOOH:

Formate

OUR:

Oxygen uptake rate

PDH:

Pyruvate dehydrogenase

pIFN-α:

Porcine interferon-α

IDH:

Isocitrate dehydrogenase

α-KGDHC:

α-Ketoglutarate dehydrogenase complex

η:

Energy efficiency

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Acknowledgments

The authors thank the financial supports from the key agricultural technology program of Shanghai Science & Technology Committee (#073919108) and Major State Basic Research Development Program (#2007CB714303), of China.

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 214122, Wuxi, People’s Republic of China

    Min-Jie Gao, Jian-Rong Wu, Zhi-Yong Zheng, Zhong-Ping Shi, Xiao-Bei Zhan & Chi-Chung Lin

  2. Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Science, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, 201106, Shanghai, People’s Republic of China

    Zhen Li & Rui-Song Yu

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  1. Min-Jie Gao
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Correspondence to Zhong-Ping Shi.

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Gao, MJ., Li, Z., Yu, RS. et al. Methanol/sorbitol co-feeding induction enhanced porcine interferon-α production by P. pastoris associated with energy metabolism shift. Bioprocess Biosyst Eng 35, 1125–1136 (2012). https://doi.org/10.1007/s00449-012-0697-1

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  • DOI: https://doi.org/10.1007/s00449-012-0697-1

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