Enzymatic production of γ-aminobutyric acid in soybeans using high hydrostatic pressure and precursor feeding.

The effects were investigated of the glutamic acid (Glu) substrate concentration on the generation and kinetics of γ-aminobutyric acid (GABA) in soybeans treated under high hydrostatic pressure (HHP; 200 MPa for 10 min at 25 °C). The conversion of Glu to GABA decreased with increasing initial Glu concentration in the soybeans. The crude glutamate decarboxylase (GAD) obtained from the HHP-treated soybeans showed substrate inhibition. The GABA production rate in the HHP-treated soybeans fitted the following substrate inhibition kinetic equation: v0=(VmaxS0)/(Km+S0+(S0)2/Ki). The Km value for the HHP-treated soybeans was significantly higher than that of the untreated soybeans. The Km values in this study show the affinity between Glu and GAD, and indicate that the HHP-treated soybeans had lower affinity between Glu and GAD than the untreated soybeans. GAD extracted from the HHP-treated soybeans showed a similar value to that in the HHP-treated soybeans. The intact biochemical system was so damaged in the HHP-treated soybeans that it showed substrate inhibition kinetics similar to that of the extracted GAD. The combination of HHP and precursor feeding proved to be a novel tool that can be used to increase the concentration of a target component.

Generation of free amino acids and gamma-aminobutyric acid in water-soaked soybean by high-hydrostatic pressure processing.

The effects of high-hydrostatic pressure processing (HPP) on soybean cotyledon as a cellular biological material were investigated from the viewpoints of the cell structure and enzyme reaction system. Damage to cell structure was evaluated by measuring dielectric properties using the Cole-Cole arc, the radius of which decreased as pressure level increased. Results suggested that cell structure was damaged by HPP. The distribution of free amino acids was measured after HPP (200 MPa) of soybean soaked in water or sodium glutamate (Glu) solution. HPP resulted in high accumulation of free amino acids in water-soaked soybean, due to proteolysis. HPP of soybean in Glu solution caused higher accumulation of gamma-aminobutyric acid, suggesting that both proteolysis and specific Glu metabolism were accelerated by HPP. We concluded that HPP partially degraded cell structure and accelerated biochemical reactions by allowing enzyme activities to remain. These events can be considered "high-pressure induced transformation" of soybean.