Expression and characterization of glutamate decarboxylase from Lactobacillus brevis HYE1 isolated from kimchi

  • Hee Seon Lim
  • Dong-Ho Seo
  • In-Tae Cha
  • Hyunjin Lee
  • Young-Do Nam
  • Myung-Ji Seo
Original Paper

Abstract

A putative gene (gadlbhye1) encoding glutamate decarboxylase (GAD) was cloned from Lactobacillus brevis HYE1 isolated from kimchi, a traditional Korean fermented vegetable. The amino acid sequences of GADLbHYE1 showed 48% homology with the GadA family and 99% identity with the GadB family from L. brevis. The cloned GADLbHYE1 was functionally expressed in Escherichia coli using inducible expression vectors. The expressed recombinant GADLbHYE1 was successfully purified by Ni–NTA affinity chromatography, and had a molecular mass of 54 kDa with optimal hydrolysis activity at 55 °C and pH 4.0. Its thermal stability was determined to be higher than that of other GADs from L. brevis, based on its melting temperature (75.18 °C). Kinetic parameters including Km and Vmax values for GADLbHYE1 were 4.99 mmol/L and 0.224 mmol/L/min, respectively. In addition, the production of gamma-aminobutyric acid in E. coli BL21 harboring gadlbhye1/pET28a was increased by adding pyridoxine as a cheaper coenzyme.

Keywords

Gamma-aminobutyric acid Glutamate decarboxylase Lactobacillus brevis Expression 

Notes

Author Contributions

All authors conceived the experiments and discussed the results. HSL and HL performed the experiments and analyzed the data with D-HS, I-TC and M-JS. D-HS, Y-DN and M-JS contributed to the writing of the paper.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

References

  1. Adeghate E, Ponery AS (2002) GABA in the endocrine pancreas: cellular localization and function in normal and diabetic rats. Tissue Cell 34:1–6.  https://doi.org/10.1054/tice.2002.0217 CrossRefGoogle Scholar
  2. Bergholz TM, Tarr CL, Christensen LM, Betting DJ, Whittam TS (2007) Recent gene conversions between duplicated glutamate decarboxylase genes (gadA and gadB) in pathogenic Escherichia coli. Mol Biol Evol 24:2323–2333.  https://doi.org/10.1093/molbev/msm163 CrossRefGoogle Scholar
  3. Biase DD, Tramonti A, John RA, Bossa F (1996) Isolation, overexpression, and biochemical characterization of the two isoforms of glutamic acid decarboxylase from Escherichia coli. Protein Expr Purif 8:430–438.  https://doi.org/10.1006/prep.1996.0121 CrossRefGoogle Scholar
  4. Capitani G, Biase DD, Aurizi C, Gut H, Bossa F, Grütter MG (2003) Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. EMBO J 22:4027–4037.  https://doi.org/10.1093/emboj/cdg403 CrossRefGoogle Scholar
  5. Casado Muñoz MdC, Benomar N, Lerma LL, Gálvez A, Abriouel H (2014) Antibiotic resistance of Lactobacillus pentosus and Leuconostoc pseudomesenteroides isolated from naturally-fermented Aloreña table olives throughout fermentation process. Int J Food Microbiol 172:110–118.  https://doi.org/10.1016/j.ijfoodmicro.2013.11.025 CrossRefGoogle Scholar
  6. Cho YR, Chang JY, Chang HC (2007) Production of gamma-aminobutyric acid (GABA) by Lactobacillus buchneri isolated from kimchi and its neuroprotective effect on neuronal cells. J Microbiol Biotechnol 17:104–109Google Scholar
  7. Cozzani I, Misuri A, Santoni C (1970) Purification and general properties of glutamate decarboxylase from Clostridium perfringens. Biochem J 118:135–141.  https://doi.org/10.1042/bj1180135 CrossRefGoogle Scholar
  8. Fagg GE, Foster AC (1983) Amino acid neurotransmitters and their pathways in the mammalian central nervous system. Neuroscience 9:701–719.  https://doi.org/10.1016/0306-4522(83)90263-4 CrossRefGoogle Scholar
  9. 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 62:689–698.  https://doi.org/10.1007/s13213-011-0307-5 CrossRefGoogle Scholar
  10. Gevers D, Huys G, Swings J (2001) Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205:31–36.  https://doi.org/10.1111/j.1574-6968.2001.tb10921.x CrossRefGoogle Scholar
  11. Ghatge MS et al (2012) Pyridoxal 5′-phosphate is a slow tight binding inhibitor of E. coli pyridoxal kinase. PLoS ONE 7:e41680.  https://doi.org/10.1371/journal.pone.0041680 CrossRefGoogle Scholar
  12. Guijin Z, Bown AW (1997) The rapid determination of γ-aminobutyric acid. Phytochemistry 44:1007–1009.  https://doi.org/10.1016/S0031-9422(96)00626-7 CrossRefGoogle Scholar
  13. Hao R, Schmit JC (1991) Purification and characterization of glutamate decarboxylase from Neurospora crassa conidia. J Biol Chem 266:5135–5139Google Scholar
  14. Hiraga K, Ueno Y, Oda K (2008a) Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate. Biosci Biotechnol Biochem 72:1299–1306.  https://doi.org/10.1271/bbb.70782 CrossRefGoogle Scholar
  15. Hiraga K, Ueno Y, Sukontasing S, Tanasupawat S, Oda K (2008b) Lactobacillus senmaizukei sp. nov., isolated from Japanese pickle. Int J Syst Evol Microbiol 58:1625–1629.  https://doi.org/10.1099/ijs.0.65677-0 CrossRefGoogle Scholar
  16. Huang J, Mei L-h, Wu H, Lin D-q (2007) Biosynthesis of γ-aminobutyric acid (GABA) using immobilized whole cells of Lactobacillus brevis. World J Microbiol Biotechnol 23:865–871.  https://doi.org/10.1007/s11274-006-9311-5 CrossRefGoogle Scholar
  17. Huang Y, Su L, Wu J (2016) Pyridoxine supplementation improves the activity of recombinant glutamate decarboxylase and the enzymatic production of γ-aminobutyric acid. PLoS ONE 11:e0157466.  https://doi.org/10.1371/journal.pone.0157466 CrossRefGoogle Scholar
  18. Inoue K, Shirai T, Ochiai H, Kasao M, Hayakawa K, Kimura M, Sansawa H (2003) Blood-pressure-lowering effect of a novel fermented milk containing γ-aminobutyric acid (GABA) in mild hypertensives. Eur J Clin Nutr 57:490–495CrossRefGoogle Scholar
  19. Jakobs C, Jaeken J, Gibson KM (1993) Inherited disorders of GABA metabolism. J Inherit Metab Dis 16:704–715.  https://doi.org/10.1007/bf00711902 CrossRefGoogle Scholar
  20. Jin Z, Mendu SK, Birnir B (2013) GABA is an effective immunomodulatory molecule. Amino Acids 45:87–94.  https://doi.org/10.1007/s00726-011-1193-7 CrossRefGoogle Scholar
  21. 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 12:707–712.  https://doi.org/10.1007/bf02931089 CrossRefGoogle Scholar
  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.  https://doi.org/10.1271/bbb.80583 CrossRefGoogle Scholar
  23. Komatsuzaki N, Shima J, Kawamoto S, Momose H, Kimura T (2005) Production of γ-aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods. Food Microbiol 22:497–504.  https://doi.org/10.1016/j.fm.2005.01.002 CrossRefGoogle Scholar
  24. 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.  https://doi.org/10.1271/bbb.70163 CrossRefGoogle Scholar
  25. Li H, Gao D, Cao Y, Xu H (2008) A high γ-aminobutyric acid-producing Lactobacillus brevis isolated from Chinese traditional paocai. Ann Microbiol 58:649–653.  https://doi.org/10.1007/bf03175570 CrossRefGoogle Scholar
  26. Li H, Qiu T, Huang G, Cao Y (2010) Production of γ-aminobutyric acid by Lactobacillus brevis NCL912 using fed-batch fermentation. Microb Cell Fact 9:85.  https://doi.org/10.1186/1475-2859-9-85 CrossRefGoogle Scholar
  27. Li H, Li W, Liu X, Cao Y (2013) gadA gene locus in Lactobacillus brevis NCL912 and its expression during fed-batch fermentation. FEMS Microbiol Lett 349:108–116.  https://doi.org/10.1111/1574-6968.12301 CrossRefGoogle Scholar
  28. Lim HS, Cha IT, Lee H, Seo MJ (2016) Optimization of γ-aminobutyric acid production by Enterococcus faecium JK29 isolated from a traditional fermented foods. Microbiol Biotechnol Lett 44:26–33CrossRefGoogle Scholar
  29. Lim HS, Cha IT, Roh SW, Shin HH, Seo MJ (2017) Enhanced production of γ-aminobutyric acid by optimizing culture conditions of Lactobacillus brevis HYE1 isolated from Kimchi: a Korean fermented food. J Microbiol Biotechnol 27:450–459.  https://doi.org/10.4014/jmb.1610.10008 CrossRefGoogle Scholar
  30. Manyam BV, Katz L, Hare TA, Kaniefski K, Tremblay RD (1981) Isoniazid-induced elevation of CSF GABA levels and effects on chorea in huntington’s disease. Ann Neurol 10:35–37.  https://doi.org/10.1002/ana.410100107 CrossRefGoogle Scholar
  31. Niesen FH, Berglund H, Vedadi M (2007) The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Nat Protoc 2:2212–2221CrossRefGoogle Scholar
  32. Okada Y, Taniguchi H, Schimada C (1976) High concentration of GABA and high glutamate decarboxylase activity in rat pancreatic islets and human insulinoma. Science 194:620–622.  https://doi.org/10.1126/science.185693 CrossRefGoogle Scholar
  33. Park K-B, Oh S-H (2007) Cloning, sequencing and expression of a novel glutamate decarboxylase gene from a newly isolated lactic acid bacterium, Lactobacillus brevis OPK-3. Bioresour Technol 98:312–319.  https://doi.org/10.1016/j.biortech.2006.01.004 CrossRefGoogle Scholar
  34. Park JY, Jeong S-J, Kim JH (2014) Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae. Biotechnol Lett 36:1791–1799.  https://doi.org/10.1007/s10529-014-1539-9 CrossRefGoogle Scholar
  35. Rizzello CG, Cassone A, Di Cagno R, Gobbetti M (2008) Synthesis of angiotensin i-converting enzyme (ACE)-inhibitory peptides and γ-aminobutyric acid (GABA) during sourdough fermentation by selected lactic acid bacteria. J Agric Food Chem 56:6936–6943.  https://doi.org/10.1021/jf800512u CrossRefGoogle Scholar
  36. Sa HD, Park JY, Jeong SJ, Lee KW, Kim JH (2015) Characterization of glutamate decarboxylase (GAD) from Lactobacillus sakei A156 isolated from Jeot-gal. J Microbiol Biotechnol 25:696–703CrossRefGoogle Scholar
  37. 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 77:853–856.  https://doi.org/10.1271/bbb.120785 CrossRefGoogle Scholar
  38. Shin S-M, Kim H, Joo Y, Lee S-J, Lee Y-J, Lee SJ, Lee D-W (2014) Characterization of glutamate decarboxylase from Lactobacillus plantarum and Its C-terminal function for the pH dependence of activity. J Agric Food Chem 62:12186–12193.  https://doi.org/10.1021/jf504656h CrossRefGoogle Scholar
  39. Siragusa S, De Angelis M, Di Cagno R, Rizzello CG, Coda R, Gobbetti M (2007) Synthesis of γ-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses. Appl Environ Microbiol 73:7283–7290.  https://doi.org/10.1128/aem.01064-07 CrossRefGoogle Scholar
  40. Tian J, Dang H, Chen Z, Guan A, Jin Y, Atkinson MA, Kaufman DL (2013) γ-Aminobutyric acid regulates both the survival and replication of human β-cells. Diabetes 62:3760–3765.  https://doi.org/10.2337/db13-0931 CrossRefGoogle Scholar
  41. Tsuchiya K, Nishimura K, Iwahara M (2003) Purification and characterization of glutamate decarboxylase from Aspergillus oryzae. Food Sci Technol Res 9:283–287.  https://doi.org/10.3136/fstr.9.283 CrossRefGoogle Scholar
  42. Yamada RH, Tsuji T, Nose Y (1977) Uptake and utilization of vitamin B6 and its phosphate esters by Escherichia coli. J Nutr Sci Vitaminol 23:7–17CrossRefGoogle Scholar
  43. 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 83:855–861.  https://doi.org/10.1002/jctb.1880 CrossRefGoogle Scholar
  44. Yu JJ, Oh SH (2011) γ-Aminobutyric acid production and glutamate decarboxylase activity of Lactobacillus sakei OPK2-59 isolated from Kimchi. Korean J Microbiol 47:316–322Google Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Life SciencesGraduate School of Incheon National UniversityIncheonRepublic of Korea
  2. 2.Research Group of Gut MicrobiomeKorea Food Research InstituteWanjuRepublic of Korea
  3. 3.Division of BioengineeringIncheon National UniversityIncheonRepublic of Korea
  4. 4.Department of Bioengineering and Nano-BioengineeringGraduate School of Incheon National UniversityIncheonRepublic of Korea

Personalised recommendations