Overexpression and optimization of glutamate decarboxylase in Lactobacillus plantarum Taj-Apis362 for high gamma-aminobutyric acid production

Microbial Biotechnology
Naser TajabadiNazamid Saari

Abstract

Gamma-aminobutyric acid (GABA) is an important bioactive compound biosynthesized by microorganisms through decarboxylation of glutamate by glutamate decarboxylase (GAD). In this study, a full-length GAD gene was obtained by cloning the template deoxyribonucleic acid to pTZ57R/T vector. The open reading frame of the GAD gene showed the cloned gene was composed of 1410 nucleotides and encoded a 469 amino acids protein. To improve the GABA-production, the GAD gene was cloned into pMG36e-LbGAD, and then expressed in Lactobacillus plantarum Taj-Apis362 cells. The overexpression was confirmed by SDS-PAGE and GAD activity, showing a 53 KDa protein with the enzyme activity increased by sevenfold compared with the original GAD activity. The optimal fermentation conditions for GABA production established using response surface methodology were at glutamic acid concentration of 497.973 mM, temperature 36°C, pH 5.31 and time 60 h. Under the conditions, maximum GABA concentration obtained (11.09 mM) was comparable with the predicted value by the model at 11.23 mM. To our knowledge, this is the first report of successful cloning (clone-back) and overexpression of the LbGAD gene from L. plantarum to L. plantarum cells. The recombinant Lactoba...Continue Reading

References

May 10, 2000·Bioscience, Biotechnology, and Biochemistry·I Kono, K Himeno
Nov 24, 2004·FEMS Microbiology Letters·M Teresa AlegreJuan M Mesas
May 10, 2007·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Sep 25, 2007·Applied and Environmental Microbiology·S SiragusaM Gobbetti
Feb 8, 2008·Bioscience, Biotechnology, and Biochemistry·Noriko KomatsuzakiJun Shima
Jan 27, 2009·International Journal of Food Microbiology·Ja Young KimKeum Taek Hwang
Apr 7, 2010·Amino Acids·Haixing Li, Yusheng Cao
Feb 12, 2014·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Naser TajabadiMohd Yazid Abdul Manap

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Citations

May 29, 2020·Bioscience, Biotechnology, and Biochemistry·Hiroki NishiokaMasanori Horie
Feb 13, 2017·Applied Microbiology and Biotechnology·Anqi ZhaoXiaoyuan Wang
Feb 8, 2020·International Journal of Molecular Sciences·Yanhua CuiXiaojun Qu
Dec 9, 2020·Microorganisms·Ida Bagus Agung YogeswaraDietmar Haltrich

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Datasets Mentioned

BETA
KF770955
AL935263
JN248358
CP002222
EF174472
EF174474
EF174473
EF174475

Methods Mentioned

BETA
PCR

Software Mentioned

esign E xpert
Stat
Clustal W

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