Optimization of γ-aminobutyric Preparation by Recombinant Glutamate Decarboxylase

doi:10.13560/j.cnki.biotech.bull.1985.2016.06.029

Abstract

Abstract: Glutamate decarboxylase(GAD)，a pyridoxal 5'-phosphate(PLP)-dependent enzyme，irreversibly catalyzes the decarboxylation of L-glutamate to be the valuable food additive γ-aminobutyric acid(GABA). In this study，a recombinant Escherichia coli BL21(DE3)/pET-24a-gad producing Lactobacillus brevis WJH3 GAD was constructed as strain in the flask culturing of fermentation and induction. The activity of GAD produced in the supernatant of culturing for 24 h medium supplemented one-time with 0.05 mmol/L PLP was 81.7 U/mL，and this was 1.8-fold of that without PLP supplementation. Furthermore，the condition for GABA preparation by enzymatic conversion was optimized；under the condition of 250 g/L monosodium glutamate(MSG)，pH5.0，37℃，60 U GAD per gram substrate incubated for 18 hours，and rotation rate 200 r/min，100% of the MSG was transformed into GABA. These results establish the utility of PLP supplementation and lay the foundation for large-scale enzymatic production of GABA.

Key words: glutamate decarboxylase, enzymatic conversion, pyridoxal 5&apos, -phosphate, monosodium glutamate, γ-aminobutyric

HUANG Yan, SU Ling-qia, WU Jing. Optimization of γ-aminobutyric Preparation by Recombinant Glutamate Decarboxylase[J]. Biotechnology Bulletin, 2016, 32(6): 199-204.

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