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Improve gamma-aminobutyric acid production in Corynebacterium glutamicum by optimizing the metabolic flux

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Abstract

Gamma-aminobutyric acid is an important nonprotein amino acid and has been extensively applied in pharmaceuticals, livestock, food additives, and so on. It is important to develop Corynebacterium glutamicum strains that can efficiently produce gamma-aminobutyric acid from glucose. In this study, production of gamma-aminobutyric acid in C. glutamicum CGY700 was improved by construction of CO2 anaplerotic reaction and overexpression of citrate synthase. The co-expression of ppc encoding phosphoenolpyruvate carboxylase and gltA encoding citrate synthase was constructed and optimized in the chromosome to compensate carbon loss and conquer metabolic bottleneck. The expression of ppc and gltA were controlled by promoters Ptac and PtacM, and the optimal mode of PtacM-ppc-Ptac-gltA was determined. Simultaneously, the genes pknG encoding serine/threonine protein kinase G and ldh encoding l-lactate dehydrogenase were deleted, and glnA2 encoding glutamine synthase was overexpressed in the chromosome. The final strain CGY-PG-304 constructed in this study could produce 41.17 g/L gamma-aminobutyric acid in shake flask cultivation and 58.33 g/L gamma-aminobutyric acid via Fed-Batch fermentation with a yield of 0.30 g/g glucose. CGY-PG-304 was constructed by genome editing; therefore, it is stable and not necessary to add any antibiotics and inducer during fermentation.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work is supported by the National Key Research and Development Program of China (2021YFC2100900) and the Key Technology Project of Inner Mongolia Autonomous Region, China (2019GG302).

Funding

Publication costs are funded by the National Key Research and Development Program of China (2021YFC2100900); the Key Technology Project of Inner Mongolia Autonomous Region, China (2019GG302).

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Authors and Affiliations

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

    Chengzhen Yao, Xiaoqing Hu & Xiaoyuan Wang

  2. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, Jiangsu, China

    Chengzhen Yao, Xiaoqing Hu & Xiaoyuan Wang

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

    Xiaoqing Hu & Xiaoyuan Wang

  4. Hulunbeier Northeast Fufeng Biotechnology Co. Ltd., Hulunbeier, China

    Yuantao Liu

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  1. Chengzhen Yao
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Contributions

XW and CY conceived and designed research. XW, CY, YL, and XH conducted experiments. XW contributed new reagents or analytical tools. XW and CY analyzed data. CY wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Xiaoyuan Wang.

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Yao, C., Liu, Y., Hu, X. et al. Improve gamma-aminobutyric acid production in Corynebacterium glutamicum by optimizing the metabolic flux. Syst Microbiol and Biomanuf 2, 305–316 (2022). https://doi.org/10.1007/s43393-021-00062-8

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  • DOI: https://doi.org/10.1007/s43393-021-00062-8

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