Article Search
닫기

Microbiology and Biotechnology Letters

보문(Article)

View PDF

Food Microbiology (FM)  |  Food Biotechnology

Microbiol. Biotechnol. Lett. 2021; 49(1): 9-17

https://doi.org/10.48022/mbl.2002.02019

Received: February 27, 2020; Accepted: April 13, 2020

Characterization of the Recombinant Glutamate Decarboxylase of Lactobacillus brevis G144 Isolated from Galchi Jeotgal, a Korean Salted and Fermented Seafood

Jeong A Kim1, Ji Yeong Park2, and Jeong Hwan Kim1,2*

1Institute of Agriculture and Life Science, 2Division of Applied Life Science BK21 4, Graduate School, Gyeongsang National University, Jinju 52828, Republic of Korea

Correspondence to :
Jeong Hwan Kim,
jeonghkm@gnu.ac.kr

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

Keywords: GABA, Lactobacillus brevis, glutamate decarboxylase, galchi jeotgal

  1. Somkuti GA, Renve JA Jr, Steinberg DH. 2012. Molecular analysis of the glutamate decarboxylase locus in Streptococcus thermophilus ST110. J. Ind. Microbiol. Biotechnol. 39: 957-963.
    Pubmed
  2. Ueno H. 2000. Enzymatic and structural aspects on glutamate decarboxylase. J. Mol. Catal. 10: 67-79.
  3. Leventhal AG, Wang Y, Pu M, Zhou Y, Ma Y. 2003. GABA and its agonists improved visual cortical function in senescent monkeys. Science 300: 812-815.
    Pubmed
  4. Li H, Cao Y. 2010. Lactic acid bacterial cell factories for γ-aminobutyric acid. Amino Acid 39: 1107-1116.
    Pubmed
  5. Park KB, Oh SH. 2007. Production of yogurt with enhanced levels of γ-aminobutyric acid and valuable nutrients using lactic acid bacteria and germinated soybean extract. Bioresour. Technol. 98:1675-1679.
    Pubmed
  6. Diana M, Quílez J, Rafecas M. 2014. γ-aminobutyric acid as a bioactive compound in foods: a review. J. Funct. Foods 10: 407-420.
  7. Cho SY, Park MJ, Kim KM, Ryu JH, Park HJ. 2011. Production of high γ-aminobutyric acid (GABA) sour kimchi using lactic acid bacteria isolated from mukeunjee kimchi. Food Sci. Biotechnol. 20: 403-408.
  8. Feehily C, Karatzas KAG. 2012. Role of glutamate metabolism in bacterial responses towards acid and other stresses. J. Appl. Microbiol. 114: 11-24.
    Pubmed
  9. Park JY, Jeong SJ, Kim JH. 2014. Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae. Biotechnol. Lett. 36: 1791-1799.
    Pubmed
  10. Sa HD, Park JY, Jeong SJ, Lee KW, Kim JH. 2015. Characterization of glutamate decarboxylase (GAD) from Lactobacillus sakei A156 isolated from Jeotgal. J. Microbiol. Biotechnol. 25: 696-703.
    Pubmed
  11. Lee KW, Shim JM, Yao Z, Kim JA, Kim HJ, Kim JH. 2017. Characterization of a glutamate decarboxylase (GAD) from Enterococcus avium M5 isolated from Jeotgal, a Korean fermented seafood. J. Microbiol. Biotechnol. 27: 1216-1222.
    Pubmed
  12. Bradford MM. 1 976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.
  13. Biase DD, Pennacchietti E. 2012. Glutamate decarboxylasedependent acid resistance in orally acquired bacteria: function, distribution and biomedical implications of the gadBC operon. Mol. Microbiol. 86: 770-786.
    Pubmed
  14. Hiraga K, Ueno Y, Oda K. 2008. Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate. Biosci. Biotechnol. Biochem. 72: 1299-1306.
    Pubmed
  15. Nomura M, Nakajima I, Fujita Y, Kobayashi M, Kimoto H, Suzuki I, et al. 1999. Lactococcus lactis contains only one glutamate decarboxylase gene. Microbiology 145: 1375-1380.
    Pubmed
  16. Nomura M, Kimoto H, Someya Y, Furukawa S, Suzuki I. 1998. Production of γ-aminobutyric acid by cheese starters during cheese ripening. J. Dairy Sci. 81: 1486-1491.
  17. Fan E, Huang J, Hu S, Mei L, Yu K. 2012. Cloning, sequencing and expression of a glutamate decarboxylase gene from the GABAproducing strain Lactobacillus brevis CGMCC1306. Ann. Microbiol. 62: 689-698.
  18. Fonda ML. 1985. L-Glutamate decarboxylase from bacteria. Methods Enzymol. 113:11-16.
  19. Lin Q, Yang S, Lu F, Lu Z, Bie X, Jiao Y, et al. 2009. Cloning and expression of glutamate decarboxylase gene from Streptococcus thermophiles Y2. J. Gen. Appl. Microbiol. 55: 305-310.
    Pubmed
  20. Nomura M, Kobayashi M, Ohmomo S, Okamoto T. 2000. Inactivation of the glutamate decarboxylase gene in Lactococcus lactis subsp. cremoris. Appl. Environ. Microbiol. 66: 2235-2237.
    Pubmed KoreaMed
  21. Seo MJ, Nam YD, Lee SY, Park SL, Yi SH, Lim SI. 2013. Expression and characterization of a glutamate decarboxylase from Lactobacillus brevis 877G producing γ-aminobutyric acid. Biosci. Biotechnol. Biochem. 77: 853-856.
    Pubmed
  22. Kim H-W, Kashima Y, Ishikawa K, Yamano N. 2009. Purification and characterization of the first archaeal glutamate decarboxylase from Pyrococcus horikoshii. Biosci. Biotechnol. Biochem. 73:224-227.
    Pubmed
  23. Tsuchiya K, Nishimura K, Iwahara M. 2003. Purification and characterization of glutamate decarboxylase from Aspergillus oryzae. Food Sci. Technol. Res. 9: 283-287.
  24. Yu J-J, Oh S-H. 2011. γ-Aminobutyric acid production and glutamate decarboxylase activity of Lactobacillus sakei OPK2-59 isolated from Kimchi. Korean J. Microbiol. 47: 316-322.
  25. Komatsuzaki N, Nakamura T, Kimura T, Shima J. 2008. Characterization of glutamate decarboxylase from a high γ-aminobutyric acid (GABA)-producer, Lactobacillus paracasei. Biosci. Biotechnol. Biochem. 72: 278-285.
    Pubmed

Starts of Metrics

Share this article on :

  • mail

Related articles in MBL

Most KeyWord ?

What is Most Keyword?

  • It is most registrated keyword in articles at this journal during for 2 years.