The taste of several bitter amino acids is reduced, sourness produced, and preference increased by gamma-glutamylization. An enzymatic method for synthesizing gamma-Glu-Val involving bacterial gamma-glutamyltranspeptidase (GGT) was developed. The optimum reaction conditions for the synthesis of gamma-Glu-Val were 20 mM Gln, 300 mM Val, and 0.04 U/ml GGT, pH 10. After 3-hr incubation at 37 degrees C, 17.6 mM gamma-Glu-Val was obtained, with the yield being 88%. gamma-Glu-Val was purified on a Dowex 1 x 8 column and then identified by NMR.