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Isolation of γ-Aminobutyric Acid Producing Lactobacillus brevis T118 from Sun-Tae Jeotgal and Its Glutamate Decarboxylase Gene CloningIsolation of γ-Aminobutyric Acid Producing Lactobacillus brevis T118 from Sun-Tae Jeotgal and Its Glutamate Decarboxylase Gene Cloning

Other Titles
Isolation of γ-Aminobutyric Acid Producing Lactobacillus brevis T118 from Sun-Tae Jeotgal and Its Glutamate Decarboxylase Gene Cloning
Authors
이세진Zhuang YaoYu MengHuong Giang Le전혜성유지연김정환
Issue Date
2020
Publisher
경상국립대학교 농업생명과학연구원
Keywords
GABA; glutamate decarboxylase; Lactobacillus brevis; gadB gene cloning
Citation
농업생명과학연구, v.54, no.4, pp 85 - 92
Pages
8
Indexed
KCI
Journal Title
농업생명과학연구
Volume
54
Number
4
Start Page
85
End Page
92
URI
https://scholarworks.gnu.ac.kr/handle/sw.gnu/7671
DOI
10.14397/jals.2020.54.4.85
ISSN
1598-5504
2383-8272
Abstract
A γ-aminobutyric acid (GABA) producing microorganism was isolated from Sun-Tae Jeotgal, a Korean traditional fermented seafood. Two thousand presumptive lactic acid bacteria (LAB) isolates were screened for GABA production by thin layer chromatography. One isolate, T118, produced GABA profusely, and identified as Lactobacillus brevis. Growth of Lb. brevis T118 was examined during 120 h cultivation in MRS broth under different conditions. Lb. brevis T118 grew well at 30-37℃, initial pH of 4-7, and up to 5% NaCl (w/v). A gene, gadB, encoding glutamate decarboxylase (GAD) was cloned by PCR. gadC encoding a glutamate/GABA antiporter was cloned and gadC located immediately upstream of gadB, indicating gadCB operon structure. The operon structure was confirmed by reverse transcription (RT)-PCR. gadB was overexpressed in Escherichia coli BL21 (DE3) and recombinant GAD was purified. The size of recombinant GAD was 54.4 kDa by SDS-PAGE, which matched well with the calculated size from the nucleotide sequence.
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