期刊
MICROBIAL CELL FACTORIES
卷 14, 期 -, 页码 -出版社
BIOMED CENTRAL LTD
DOI: 10.1186/s12934-015-0205-9
关键词
Corynebacterium glutamicum; Gamma-aminobutyrate; Glutamate; Glutamate decarboxylase; Biotin; Fed-batch cultivation
资金
- Technology Development Program to Solve Climate Changes (Systems Metabolic Engineering for Biorefineries) [NRF-2012-C1AAA001-2012M1A2A2026556]
- Intelligent Synthetic Biology Center of Global Frontier Project from the Ministry of Science, ICT and Future Planning (MSIP) through the National Research Foundation (NRF) of Korea [NRF-2014M3A6A8066443]
- Industrial Strategic Technology Development Program (Production of biobased cadaverine and polymerization of Bio-polyamide 510) - Ministry of Trade, Industry & Energy (MOTEI, Korea) [10047910]
Background: Gamma-aminobutylate (GABA) is an important chemical in pharmacetucal field and chemical industry. GABA has mostly been produced in lactic acid bacteria by adding L-glutamate to the culture medium since L-glutamate can be converted into GABA by inherent L-glutamate decarboxylase. Recently, GABA has gained much attention for the application as a major building block for the synthesis of 2-pyrrolidone and biodegradable polyamide nylon 4, which opens its application area in the industrial biotechnology. Therefore, Corynebacterium glutamicum, the major L-glutamate producing microorganism, has been engineered to achieve direct fermentative production of GABA from glucose, but their productivity was rather low. Results: Recombinant Corynebacterium glutamicum strains were developed for enhanced production of GABA from glucose by expressing Escherichia coli glutamate decarboxylase (GAD) mutant, which is active in expanded pH range. Synthetic P-H36, P-I16, and P-L26 promoters, which have different promoter strengths in Corynebacterium glutamicum, were examined for the expression of Escherichia coli GAD mutant. Corynebacterium glutamicum expressing Escherichia coli GAD mutant under the strong PH36 promoter could produce GABA to the concentration of 5.89 +/- 0.35 g/L in GP1 medium at pH 7.0, which is 17-fold higher than that obtained by Corynebacterium glutamicum expressing wild-type Escherichia coli GAD in the same condition (0.34 +/- 0.26 g/L). Fed-bath culture of Corynebacterium glutamicum expressing Escherichia coli GAD mutant in GP1 medium containing 50 mu g/L of biotin at pH 6, culture condition of which was optimized in flask cultures, resulted in the highest GABA concentration of 38.6 +/- 0.85 g/L with the productivity of 0.536 g/L/h. Conclusion: Recombinant Corynebacterium glutamicum strains developed in this study should be useful for the direct fermentative production of GABA from glucose, which allows us to achieve enhanced production of GABA suitable for its application area in the industrial biotechnology.
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