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
- 이세진; Zhuang Yao; Yu Meng; Huong Giang Le; 전혜성; 유지연; 김정환
- 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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