Preparation and Kokumi Properties of N-Acetyl-Val/Leu/Ile/Met/Phe in the Presence of Acetic Acid and Amino Acid: A Commercially Available Transglutaminase and Protease A 2SD

Kokumi is a beneficial feeling for the evaluation of food quality, and thus, preparing and understanding the taste properties of kokumi compounds are important for the flavor of food. N-acetyl-Val/Leu/Ile/Met/Phe/Trp/Tyr is a type of kokumi compound found in food and usually prepared by chemical reagents. In this study, we first prepared these six kokumi compounds using transglutaminase and protease A2SD in aqueous solution by using amino acids and acetic acid as substrates and evaluated their kokumi characteristics. HPLC and LC-MS were used to identify quantitative N-acetyl amino acids. Using Phe and acetic acid as substrates, transglutaminase and protease A2SD showed the highest yields for N-acetyl-Phe of 22.75 and 42.21%, respectively, under the optimal conditions. For N-acetyl-Val/Leu/Ile/Met/Trp/Tyr, these two enzymes showed the synthesis yield in the ranges of 2.22-20.12 and 0.75-12.91%, respectively. Six N-acetyl-amino acids were succesully enriched by ethyl acetate with a recovery over 50% and purity over 95%. Sensory evaluation found that N-acetyl-Val/Leu/Ile/Met/Phe are kokumi compounds that enhance sweet, umami, and salt tastes in 5% sucrose, 0.3% NaCl, and 0.5% sodium glutamate, especially N-acetyl-Val, with the salt- and umami-enhancing threshold values of 0.63 and 1.25 g/L, respectively. Therefore, transglutaminase and protease A2SD for the synthesis of partial N-acetyl amino acid might have the potential to be applied in food as a kokumi compound.