Title |
Heterologous expression and enzymatic characterization of γ-glutamyltranspeptidase from Bacillus amyloliquefaciens |
Author |
Jung-Min Lee1, Jaejung Lee1,2, Gyeong-Hwa Nam1, Byung-Sam Son1, Myoung-Uoon Jang1, So-Won Lee2, Byung-Serk Hurh2, and Tae-Jip Kim1* |
Address |
1Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University, Cheongju 28644, Republic of Korea, 2Sempio Fermentation Research Center, Sempio Foods Company, Cheongju 28156, Republic of Korea |
Bibliography |
Journal of Microbiology, 55(2),147-152, 2017,
|
DOI |
10.1007/s12275-017-6638-6
|
Key Words |
γ-glutamyltranpeptidase, Bacillus amyloliquefaciens,
heterologous expression, enzymatic characterization |
Abstract |
γ-Glutamyltranspeptidase (GGT) catalyzes the cleavage of γ-
glutamyl compounds and the transfer of γ-glutamyl moiety
to water or to amino acid/peptide acceptors. GGT can be utilized
for the generation of γ-glutamyl peptides or glutamic
acid, which are used as food taste enhancers. In the present
study, Bacillus amyloliquefaciens SMB469 with high GGT activity
was isolated from Doenjang, a traditional fermented soy
food of Korea. The gene encoding GGT from B. amyloliquefaciens
SMB469 (BaGGT469) was cloned from the isolate, and
heterologously expressed in E. coli and B. subtilis. For comparison,
three additional GGT genes were cloned from B.
subtilis 168, B. licheniformis DSM 13, and B. amyloliquefaciens
FZB42. The BaGGT469 protein was composed of 591
amino acids. The final protein comprises two separate polypeptide
chains of 45.7 and 19.7 kDa, generated via autocatalytic
cleavage. The specific activity of BaGGT469 was determined
to be 17.8 U/mg with γ-L-glutamyl-p-nitroanilide
as the substrate and diglycine as the acceptor. GGTs from B.
amyloliquefaciens showed 1.4- and 1.7-fold higher transpeptidase
activities than those from B. subtilis and B. licheniformis,
respectively. Especially, recombinant B. subtilis expressing
BaGGT469 demonstrated 11- and 23-fold higher GGT
activity than recombinant E. coli and the native B. amyloliquefaciens,
respectively, did. These results suggest that BaGGT469
can be utilized for the enzymatic production of various γ-
glutamyl compounds. |