CIESC Journal ›› 2016, Vol. 67 ›› Issue (5): 2015-2021.doi: 10.11949/j.issn.0438-1157.20151432

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Kinetic modelling of porcine insulin precursor (PIP) expressed by multi-copy recombinant Pichia pastoris

CHEN Li1,2, WANG Yue2, GUO Meijin2, CHU Ju2, ZHUANG Yingping2   

  1. 1 National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, China;
    2 Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, Shanghai 200237, China
  • Received:2015-09-10 Revised:2015-11-19 Online:2016-05-05 Published:2015-11-23
  • Supported by:

    supported by the Special Fund for Agro-scientific Research in the Public Interest(201303046), the National High Technology Research and Development Program of China (2012AA021201), and the Open Project Program of State Key Laboratory of Bioreactor Engineering (2060204).

Abstract:

Kinetic modelling of recombinant Pichia pastoris harboring multiple porcine insulin precursor (PIP) gene dosages was studied when the cells were grown in the fed-batch culture. The key parameters of this kinetics were estimated, including specific cell growth rate (h-1), PIP production rate (g·g-1·h-1) and substrate consumption rate (g·g-1·h-1) with nonlinear curve fit by Origin8.0. The results showed that both growth-associated production coefficient ( ) and growth-associated metabolism coefficient (k1) increased with increasing copy numbers. The expression level of PIP reached the highest at the copy number of 12. These results suggested that rapid growth and lower metabolic burden of a high copy number effectively improved the production rate of target proteins. Furthermore, the predicted values based on the established kinetic model were in good agreement with the experimental data, indicating that the kinetic model could be used to describe recombinant PIP production process in fed-batch fermentation mode.

Key words: recombinant Pichia pastoris, porcine insulin precursor expression, gene dosage, kinetic model

CLC Number: 

  • TQ464.7

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