GLUTAMATE DECARBOXYLASE GENE FROM LACTIC ACID BACTERIA OF LACTOBACILLUS BREVIS OPK-3

摘要

A glutamate decarboxylase gene isolated from lactic acid bacterium Lactobacillus brevis OPK-3(KFCC 11330) is provided to produce the high concentration of GABA(gamma amino butyric acid). The glutamate decarboxylase(GAD) gene isolated from lactic acid bacterium Lactobacillus brevis OPK-3(KFCC 11330) has the nucleotide sequence of SEQ ID NO:3. A transformed Escherichia coli BL21(DE3) is produced by transforming Escherichia coli with a recombinant vector, pTYB2/GAD, and containing the glutamate decarboxylase(GAD) gene isolated from Lactobacillus brevis OPK-3(KFCC 11330). A method for cloning the glutamate decarboxylase(GAD) comprises the steps of: PCR-amplifying the glutamate decarboxylase(GAD) gene with a primer to obtain the DNA fragments of GAD, wherein the primer is obtained from the glutamate decarboxylase of Lactobacillus brevis OPK-3(KFCC 11330) and has 70% or more homology to the nucleotide sequences of Lactococcus lactis GAD and Lactobacillus plantarum GAD; ligating the DNA fragment of GAD to a cloning vector pGEM T-Easy and transforming it; and ligating the transformed DNA fragments of GAD to an expression vector pTYB2 and transforming it.