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A multistrategy approach for improving the expression of E. coli phytase in Pichia pastoris

  • Genetics and Molecular Biology of Industrial Organisms - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Phytase is an additive in animal feed that degrades phytic acid in plant material, reducing feeding costs, and pollution from fecal phosphorus excretion. A multistrategy approach was adopted to improve the expression of E. coli phytase in Pichia pastoris. We determined that the most suitable signal peptide for phytase secretion was an α-factor secretion signal with an initial enzyme activity of 153.51 U/mL. Increasing the copy number of this gene to four increased phytase enzyme activity by 234.35%. PDI overexpression and Pep4 gene knockout increased extracellular phytase production by 35.33% and 26.64%, respectively. By combining favorable factors affecting phytase expression and secretion, the enzyme activity of the phytase-engineered strain was amplified 384.60% compared with that of the original strain. We also evaluated the potential for the industrial production of the engineered strain using a 50-L fed-batch fermenter and achieved a total activity of 30,246 U/mL after 180 h of fermentation.

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Acknowledgements

This work was supported by the National Key R&D Program of China (2018YFD0901001) and the Tianjin Science Fund for Distinguished Young Scholars (17JCJQJC45300). We thank Ms. Jie Zhang from TIB-CAS for her help with the enzyme activity determination.

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  1. Yuankun Helian and Yuanming Gai contributed equally to this work.

Authors and Affiliations

  1. School of Biological Engineering, Dalian Polytechnic University, No. 1 Qinggongyuan, Ganjingzi, Dalian, 116034, Liaoning, People’s Republic of China

    Yuankun Helian & Yumei Sun

  2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, People’s Republic of China

    Yuankun Helian, Yuanming Gai, Huan Fang & Dawei Zhang

  3. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, People’s Republic of China

    Yuankun Helian, Yuanming Gai, Huan Fang & Dawei Zhang

  4. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Dawei Zhang

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  1. Yuankun Helian
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Helian, Y., Gai, Y., Fang, H. et al. A multistrategy approach for improving the expression of E. coli phytase in Pichia pastoris. J Ind Microbiol Biotechnol 47, 1161–1172 (2020). https://doi.org/10.1007/s10295-020-02311-6

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