PROCESS DEVELOPMENT FOR Γ-AMINOBUTYRIC ACID (GABA) PRODUCTION BY BIOTRANSFORMATION
Abstract
Gamma-amino butyric acid (GABA), a neurotransmitter in humans’ brains, has several physiological functions such as relieving anxiety, depression, and insomnia as well as having an anti-stress effect in humans. In recent years, the trend of adding GABA to food and drink products has been popular, especially in Japan; therefore, increased GABA production is required to support commercial demand. The objective of this study was to screen the GABA producing bacteria which could bio-transform glutamate into GABA via the glutamate decarboxylase (GAD) enzyme with whole cell reaction. Furthermore, the culture media and culture conditions that can promote high cell growth and GAD enzyme activity, as well as the process optimization, were investigated. The results showed that L. brevis had the highest GABA production capability (1.3 g/L). It was found that the GAD activity of the cell depended on growth as the activity was high during the mid-log and decreased during the stationary phase. The reaction was enhanced by increasing the temperature up to 40oC and adding Pyridoxal 5’-phosphate (PLP) coenzyme (the obtained productivity, final GABA concentration, and yield were 3.4 mM/h, 2.7 g GABA/L, and 0.94 mol GABA/mol glutamate, respectively). For the biotransformation reaction, it was found that the GAD activity dropped with reuse of the whole cell. The temperature at 30oC and pH in the range 3.0-5.0 were the optimal conditions for the GABA reaction. However, the GABA reaction with the cell recycling technique required a PLP addition every cycle. α-ketoglutarate was also tested as a substitute for PLP regeneration but it could not enter into the cell. The alternative process for GABA production was growth-associated production which could avoid cell death during cell reaction and showed better GABA production performance than that of cell reaction (5.1 mM/h, 16 g GABA /L, and 0.90 mol GABA/mol glutamate of yield, respectively).
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