Biotechnology Letters

, Volume 36, Issue 9, pp 1791–1799 | Cite as

Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae

  • Ji Yeong Park
  • Seon-Ju Jeong
  • Jeong Hwan KimEmail author
Original Research Paper


Lactic acid bacteria (LAB) were isolated from Kimchi, a Korean traditional fermented vegetable food. LAB accumulating GABA (γ-aminobutyric acid) in the culture media were screened by TLC analysis. One isolate, GU240, produced the highest amount of GABA among the 3,000 isolates and identified as a Lactobacillus zymae strain. Glutamate decarboxylase (GAD) gene was cloned and over-expressed in E. coli BL21(DE3) using pET26b(+). The recombinant GAD was purified by using a Ni–NTA column. Its size was 53 kDa by SDS-PAGE. Maximum GAD activity was at pH 4.5 and 41 °C and the activity was dependent on pyridoxal 5′-phosphate. Km and Vmax of LzGAD were 1.7 mM and 0.01 mM/min, respectively, when glutamate was used as a substrate.


γ-Aminobutyric acid gad gene cloning Glutamate decarboxylase Lactobacillus zymae 



This work was supported by Geochang HanSsal and Geochang county Regional Agriculture Specialization fund. J. Y. Park and S.-J. Jeong were supported by BK21 Plus from MOE, Republic of Korea.


  1. Fan E, Huang J, Hu S, Mei L, Yu K (2012) Cloning, sequencing and expression of a glutamate decarboxylase gene from the GABA-producing strain Lactobacillus brevis CGMCC 1306. Ann Microbiol 62:689–698CrossRefGoogle Scholar
  2. Fonda ML (1985) l-Glutamate decarboxylase from bacteria. Methods Enzymol 113:11–16PubMedCrossRefGoogle Scholar
  3. Higuchi T, Hayashi H, Abe K (1997) Exchange of glutamate and gamma-aminobutyrate in a Lactobacillus strain. J Bacteriol 179:3362–3364PubMedCentralPubMedGoogle Scholar
  4. Hiraga K, Ueno Y, Oda K (2008) Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate. Biosci Biotechnol Biochem 72:1299–1306PubMedCrossRefGoogle Scholar
  5. Huang J, Mei L, Sheng Q, Yao S, Lin D (2007) Purification and characterization of glutamate decarboxylase of Lactobacillus brevis CGMCC 1306 isolated from fresh milk. Chin J Chem Eng 15:157–161CrossRefGoogle Scholar
  6. Johnson BS, Singh NK, Cherry JH, Locy RD (1997) Purification and characterization of glutamate decarboxylase from cowpea. Phytochemistry 46:39–44CrossRefGoogle Scholar
  7. Kim S-H, Shin B-H, Kim Y-H, Nam S-W, Jeon S-J (2007) Cloning and expression of a full-length glutamate decarboxylase gene from Lactobacillus brevis BH2. Biotechnol Bioprocess Eng 12:707–712CrossRefGoogle Scholar
  8. 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 72:278–285PubMedCrossRefGoogle Scholar
  9. Li H, Cao Y (2010) Lactic acid bacterial cell factories for gamma-aminobutyric acid. Amino Acid 39:1107–1116CrossRefGoogle Scholar
  10. Lin Q, Yang S, Lü F, Lu Z, Bie X, Jiao Y, Zou X (2009) Cloning and expression of glutamate decarboxylase gene from Streptococcus thermophilus Y2. J Gen Appl Microbiol 55:305–310PubMedCrossRefGoogle Scholar
  11. Murzin AG (1996) Structural classification of proteins: new superfamilies. Curr Opin Struct Biol 6:386–394PubMedCrossRefGoogle Scholar
  12. Nomura M, Nakajima I, Fujita Y, Kobayashi M, Kimoto H, Suzukil I, Aso H (1999) Lactococcus lactis contains only one glutamate decarboxylase gene. Microbiology 145:1375–1380PubMedCrossRefGoogle Scholar
  13. Park KB, Oh SH (2007) Cloning, sequencing and expression of a novel glutamate decarboxylase gene from a newly isolated lactic acid bacterium, Lactobacillus brevis OPK-3. Bioresour Technol 98:312–319PubMedCrossRefGoogle Scholar
  14. Seo M-J, Nam Y-D, Lee S-Y, Park S-L, Yi S-H, Lim S-I (2013) Expression and characterization of a glutamate decarboxylase from Lactobacillus brevis 877G producing γ-aminobutyric acid. Biosci Biotechnol Biochem 77:853–856PubMedCrossRefGoogle Scholar
  15. Somkuti GA, Renve JA Jr, Steinberg DH (2012) Molecular analysis of the glutamate decarboxylase locus in Streptococcus thermophilus ST110. J Ind Microbiol Biotechnol 39:957–963PubMedCrossRefGoogle Scholar
  16. Ueno Y, Hayakawa K, Takahashi S, Oda K (1997) Purification and characterization of glutamate decarboxylase from Lactobacillus brevis IFO 12005. Biosci Biotechnol Biochem 61:1168–1171PubMedCrossRefGoogle Scholar
  17. Vancanneyt M, Neysens P, De Wachter M, Engelbeen K, Snauwaert C, Cleenwerck I, Meulen RVd, Hoste B, Tsakalidou E, Vuyst LD, Swings J (2005) Lactobacillus acidifarinae sp. nov. and Lactobacillus zymae sp. nov., from wheat sourdoughs. Int J Syst Evol Microbiol 55:615–620PubMedCrossRefGoogle Scholar
  18. Wang Q, Xin Y, Zhang F, Feng Z, Fu J, Luo L, Yin Z (2011) Enhanced γ-aminobutyric acid-forming activity of recombinant glutamate decarboxylase (gadA) from Escherichia coli. World J Microbiol Biotechnol 27:693–700CrossRefGoogle Scholar
  19. Yang S-Y, Lin Q, Lu Z-X, Lü F-X, Bie X-M, Zou X-K, Sun L-J (2008) Characterization of a novel glutamate decarboxylase from Streptococcus salivarius ssp. thermophilus Y2. J Chem Technol Biotechnol 83:855–861CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Ji Yeong Park
    • 1
  • Seon-Ju Jeong
    • 1
  • Jeong Hwan Kim
    • 1
    Email author
  1. 1.Division of Applied Life Science (BK21 Plus), Graduate School, Institute of Agriculture and Life ScienceGyeongsang National UniversityJinjuRepublic of Korea

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