Effects of overexpression of carboxylation pathway genes and inactivation of malic enzymes on malic acid production in Escherichia coli

Fei Lou1,2 Ning Li1,2 Yujiao Zhao1,2 Shiting Guo1,2 Zhiwen Wang1,2 Tao Chen1,2

(1.Key Laboratory of Systems Bioengineering, Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin , China 300072)
(2.Collaborative Innovation Center of Chemical Science and Engineering, Tianjin , China 300072)

【Abstract】Malic acid is a dicarboxylic acid that is widely used in food, pharmaceutical and chemical industries. We studied the effects of overexpression of carboxylation pathway genes and inactivation of malic enzymes on the aerobic production of malic acid. Over expression of phosphoenolpyruvate (PEP) carboxylase (ppc) generated strain E21, which increased malic acid production from 0.57 g/L to 3.83 g/L. Then pyc gene from Coryenbacterium glutamicus and pck gene from Actinobacillus succinogenes were overexpressed in E21 separately. The resulting strains E21 (pTrcpyc) and E21 (pTrc-A-pck) produced 6.04 and 5.01 g/L malate with a yield of 0.79 and 0.65 mol/mol glucose, respectively. Deleting two malic enzymes (encoded by maeA and maeB) also led to an increase of 36% in malic acid production with a production of 5.21 g/L. However, the combination of malic enzymes deletion and pyc overexpression could not further increase the yield of malic acid. After optimization of fermentation conditions, strain E21 (pTrcpyc) produced 12.45 g/L malic acid with a yield of 0.84 mol/mol which is 63.2% of the theoretical yield.

【Keywords】 Escherichia coli; metabolic engineering; malic acid; malic enzyme; carboxylation pathway;

【DOI】

【Funds】 National Natural Science Foundation of China (No. NSFC-21390201) National Natural Science Foundation of China(No.NSFC-21390201) National Basic Research Program of China (973 Program) (No. 2012CB725203) National Basic Research Program of China(973 Program)(No.2012CB725203)

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(Translated by ZHAO Bo)

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This Article

ISSN:1000-3061

CN: 11-1998/Q

Vol 32, No. 11, Pages 1539-1548

November 2016

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Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Conclusions
  • References