Identification and Activity Characterization of γ-Glutamyltransferase from Bovine Milk.

Abstract

It is well known that bovine milk contains γ-glutamyltransferase (GGT) activity. To verify the identity of the GGT and further to characterize the generation of γ-glutamyl peptides, identification of GGT from bovine milk and quantification of kokumi peptides and free amino acids were performed. GGT was purified from skim milk and identified as the bovine protein (G3N2D8), and it reveals that it is composed of two subunits. Sequence alignment with human GGT and molecular mass determination showed that the bovine GGT was glycosylated and contained an N-terminal transmembrane part. Further activity characterization was performed in comparison with GGT from Bacillus amyloliquefaciens in terms of the ability to generate γ-glutamyl peptides from casein hydrolysates. During the transpeptidation reaction catalyzed by both GGT, γ-glutamyl peptides significantly (P < 0.05) increased after γ-glutamylation; addition of glutamine contributed to the generation of γ-glutamyl peptides, suggesting that glutamine could act as a γ-glutamyl donor. This study reveals that the GGT of skim milk membranes is a glycosylated membrane protein that can generate γ-glutamyl peptides.