Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (2): 138-148.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0529

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Expression and Fermentation Optimization of Candida antarctica Lipase B in Escherichia coli

WU Rong1(), CAO Jia-rui1, CAO Jun1, LIU Fei-xiang1, YANG Meng1, SU Er-zheng1,2()   

  1. 1. College of Light Industry and Food Engineering,Nanjing Forestry University,Nanjing 210037
    2. Co-innovation Center for the Sustainable Forestry in Southern China,Nanjing Forestry University,Co-Innovation Center for Efficient Processing and Utilization of Forest Products,Nanjing Forestry University,Nanjing 210037
  • Received:2020-05-25 Online:2021-02-26 Published:2021-02-26
  • Contact: SU Er-zheng E-mail:rongwu@njfu.edu.cn;ezhsu@njfu.edu.cn

Abstract:

In order to achieve efficient soluble expression of Candida antarctica lipase B(CALB)in Escherichia coli at lower cost,CALB gene expression plasmids with different signal peptides were constructed and transformed into different E. coli hosts,and then a series of fermentation experiments were employed in shaking flasks to optimize the culture medium types,inducing conditions,components of medium and time course of cultivation. The results showed that the recombinant strain pET25b-CALB-1/Rosetta(DE3)with PelB signal peptide was induced to have the best expression in TY medium at 20℃ employing 0.5%(W/V)lactose. The optimal composition of fermentation medium was 1.75%(W/V)sorbitol,2.25%(W/V)fish peptone,1%(W/V)Angel yeast extract and 0.75%(W/V)Na2HPO4. The recombination system achieved a maximum CALB activity of 35.67 U/mL after induction for 60 h,which was 17.77 times higher than that before optimization,i.e.,the highest one of CALB produced by E. coli at present. In conclusion,the expression system in E. coli was successfully established,and the high-level soluble expression of CALB was obtained after system optimization.

Key words: Candida antarctica lipase B, Escherichia coli, soluble expression, fermentation optimization