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Enhancement of γ-aminobutyric acid production in recombinant Corynebacterium glutamicum by co-expressing two glutamate decarboxylase genes from Lactobacillus brevis

  • Metabolic Engineering and Synthetic Biology
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

γ-Aminobutyric acid (GABA), a non-protein amino acid, is a bioactive component in the food, feed and pharmaceutical fields. To establish an effective single-step production system for GABA, a recombinant Corynebacterium glutamicum strain co-expressing two glutamate decarboxylase (GAD) genes (gadB1 and gadB2) derived from Lactobacillus brevis Lb85 was constructed. Compared with the GABA production of the gadB1 or gadB2 single-expressing strains, GABA production by the gadB1–gadB2 co-expressing strain increased more than twofold. By optimising urea supplementation, the total production of l-glutamate and GABA increased from 22.57 ± 1.24 to 30.18 ± 1.33 g L−1, and GABA production increased from 4.02 ± 0.95 to 18.66 ± 2.11 g L−1 after 84-h cultivation. Under optimal urea supplementation, l-glutamate continued to be consumed, GABA continued to accumulate after 36 h of fermentation, and the pH level fluctuated. GABA production increased to a maximum level of 27.13 ± 0.54 g L−1 after 120-h flask cultivation and 26.32 g L−1 after 60-h fed-batch fermentation. The conversion ratio of l-glutamate to GABA reached 0.60–0.74 mol mol−1. By co-expressing gadB1 and gadB2 and optimising the urea addition method, C. glutamicum was genetically improved for de novo biosynthesis of GABA from its own accumulated l-glutamate.

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Acknowledgments

The authors thank the Program of State Key Laboratory of Food Science and Technology (contract no. SKLF-TS-201103) and the Fundamental Research Funds for the Central Universities (contract no. JUSRP 21109) for financial support.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China

    Feng Shi, Junjun Jiang, Youxin Li & Yilong Xie

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

    Feng Shi, Junjun Jiang, Youxin Li & Yilong Xie

  3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China

    Yongfu Li

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  1. Feng Shi
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  2. Junjun Jiang
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Correspondence to Feng Shi.

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F. Shi and J. Jiang contributed equally to this work.

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Shi, F., Jiang, J., Li, Y. et al. Enhancement of γ-aminobutyric acid production in recombinant Corynebacterium glutamicum by co-expressing two glutamate decarboxylase genes from Lactobacillus brevis . J Ind Microbiol Biotechnol 40, 1285–1296 (2013). https://doi.org/10.1007/s10295-013-1316-0

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  • DOI: https://doi.org/10.1007/s10295-013-1316-0

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