大连工业大学学报2025，Vol.44Issue(2)：98-102,5.DOI:10.19670/j.cnki.dlgydxxb.2025.0204
短乳杆菌NM-1谷氨酸脱羧酶基因的克隆表达及其酶活力
Gene cloning and expression of glutamic acid decarboxylase and its activity from Levilactobacillus brevis NM-1
摘要
Abstract
Glutamate decarboxylase(GAD)can convert the substrate L-glutamic acid(L-Glu)intoγ-aminobutyric acid(GABA),and the enzyme activity of GAD is a key factor affecting the efficiency of GABA synthesis.To increase the production of GABA,a lactic acid bacteria producing glutamic acid decarboxylase was screened from nature and preliminarily identified as Levilactobacillus brevis,named Levilactobacillus brevis NM-1.The GAD encoding genes gadA and gadB in Levilactobacillus brevis NM-1 were cloned and ligated into the pET-30a(+)expression plasmid,then the recombinant plasmids were transferred into E.coli BL21(DE3)by heat shock to create the genetically engineered E.coli BL21(DE3)/pET-30a(+)-gadA and E.coli BL21(DE3)/pET-30a(+)-gadB(GadA-30a and GadB-30a).The results showed that the whole cell catalytic capacity of GadB-30a was much higher than that of GadA-30a at the same culture conditions,suggesting that the GAD activity of Levilactobacillus brevis NM-1 depending on the gadB gene.The GAD activity of GadB-30a was 6.52 U/mL and the specific enzyme activity was 9.59 U/mg.关键词
谷氨酸脱羧酶/短乳杆菌/L-谷氨酸/γ-氨基丁酸Key words
glutamate decarboxylase/Levilactobacillus brevis/L-glutamic acid/γ-aminobutyric acid分类
生物学引用本文复制引用
李文杰,谭琛,刘昊,张春枝..短乳杆菌NM-1谷氨酸脱羧酶基因的克隆表达及其酶活力[J].大连工业大学学报,2025,44(2):98-102,5.基金项目
辽宁省教育厅高等学校基本科研项目(重点项目)(LJIKZZ20220060). （重点项目）
