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Improving Performance and Operational Stability of Porcine Interferon-α Production by Pichia pastoris with Combinational Induction Strategy of Low Temperature and Methanol/Sorbitol Co-feeding

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Abstract

Various induction strategies were investigated for effective porcine interferon-α (pIFN-α) production by Pichia pastoris in a 10 L fermenter. We found that pIFN-α concentration could be significantly improved with the strategies of low-temperature induction or methanol/sorbitol co-feeding. On this basis, a combinational strategy of induction at lower temperature (20 °C) with methanol/sorbitol co-feeding has been proposed for improvement of pIFN-α production. The results reveal that maximal pIFN-α concentration and antiviral activity reach the highest level of 2.7 g/L and 1.8 × 107 IU/mg with the proposed induction strategy, about 1.3–2.1 folds higher than those obtained with other sub-optimal induction strategies. Metabolic analysis and online multi-variable measurement results indicate that energy metabolic enrichment is responsible for the performance enhancement of pIFN-α production, as a large amount of ATP could be simultaneously produced from both formaldehyde oxidation pathway in methanol metabolism and tricarboxylic acid (TCA) cycle in sorbitol metabolism. In addition, the proposed combinational induction strategy enables P. pastoris to be resistant to high methanol concentration (42 g/L), which conceivably occur associating with the error-prone methanol over-feeding. As a result, the proposed combinational induction strategy simultaneously increased the targeted protein concentration and operational stability leading to significant improvement of pIFN-α production.

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Acknowledgments

The authors thank the financial supports from National Natural Science Foundation of China (#31301408), Natural Science Foundation of Jiangsu Province (#BK20130122), China Postdoctoral Science Foundation (#2014 M551501), a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions,the 111 Project (No. 111-2-06),and the Jiangsu province “Collaborative Innovation Center for Advanced Industrial Fermentation” industry development program.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

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

    Min-Jie Gao, Xiao-Bei Zhan, Peng Gao, Xu Zhang, Zhong-Ping Shi & Chi-Chung Lin

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

    Shi-Juan Dong & Zhen Li

Authors
  1. Min-Jie Gao
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  3. Peng Gao
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Correspondence to Xiao-Bei Zhan or Chi-Chung Lin.

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Gao, MJ., Zhan, XB., Gao, P. et al. Improving Performance and Operational Stability of Porcine Interferon-α Production by Pichia pastoris with Combinational Induction Strategy of Low Temperature and Methanol/Sorbitol Co-feeding. Appl Biochem Biotechnol 176, 493–504 (2015). https://doi.org/10.1007/s12010-015-1590-6

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