Biotechnological advances and perspectives of gamma-aminobutyric acid production



Gamma-aminobutyric acid (GABA) is a four-carbon non-protein amino acid that is widely distributed among various organisms. Since GABA has several well-known physiological functions, such as mediating neurotransmission and hypotensive activity, as well as having tranquilizer effects, it is commonly used as a bioactive compound in the food, pharmaceutical and feed industries. The major pathway of GABA biosynthesis is the irreversible decarboxylation of l-glutamate catalyzed by glutamate decarboxylase (GAD), which develops a safe, sustainable and environmentally friendly alternative in comparison with traditional chemical synthesis methods. To date, several microorganisms have been successfully engineered for high-level GABA biosynthesis by overexpressing exogenous GADs. However, the activity of almost all reported microbial GADs sharply decreases at physiological near-neutral pH, which in turn provokes negative effects on the application of these GADs in the recombinant strains for GABA production. Therefore, ongoing efforts in the molecular evolution of GADs, in combination with high-throughput screening and metabolic engineering of particular producer strains, offer fascinating new prospects for effective, environmentally friendly and economically viable GABA biosynthesis. In this review, we briefly introduce the applications in which GABA is used, and summarize the most important methods associated with GABA production. The major achievements and present challenges in the biotechnological synthesis of GABA, focusing on screening and enzyme engineering of GADs, as well as metabolic engineering strategy for one-step GABA biosynthesis, will be extensively discussed.


GABA biosynthesis Glutamate decarboxylase Enzyme engineering Metabolic engineering 



This study was supported by the National Natural Science Foundation of China (No. 31500044) and the “Hundred Talents Program” of the Chinese Academy of Sciences.


  1. Ab Kadir S, Wan-Mohtar WA, Mohammad R et al (2016) Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for gamma-aminobutyric acid (GABA) production. J Ind Microbiol Biotechnol 43:1387–1395CrossRefGoogle Scholar
  2. Alcalde M, Ferrer M, Plou FJ (2007) Environmental biocatalysis: from remediation with enzymes to novel green processes. Biocatal Biotransfo 25:113–113CrossRefGoogle Scholar
  3. Aoki H, Uda I, Tagami K et al (2003) The production of a new tempeh-like fermented soybean containing a high level of gamma-aminobutyric acid by anaerobic incubation with Rhizopus. Biosci Biotech Bioch 67:1018–1023CrossRefGoogle Scholar
  4. Awapara J, Landua AJ, Fuerst R, Seale B (1950) Free gamma-aminobutyric acid in brain. J Biol Chem 187:35–39Google Scholar
  5. Bouche N, Fromm H (2004) GABA in plants: just a metabolite? Trends Plant Sci 9:110–115CrossRefGoogle Scholar
  6. Capitani G, De Biase D, Aurizi C et al (2003) Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. Embo J 22:4027–4037CrossRefGoogle Scholar
  7. Choi JW, Yim SS, Lee SH et al (2015) Enhanced production of gamma-aminobutyrate (GABA) in recombinant Corynebacterium glutamicum by expressing glutamate decarboxylase active in expanded pH range. Microb Cell Fact 14:21CrossRefGoogle Scholar
  8. Coleman ST, Fang TK, Rovinsky SA et al (2001) Expression of a glutamate decarboxylase homologue is required for normal oxidative stress tolerance in Saccharomyces cerevisiae. J Biol Chem 276:244–250CrossRefGoogle Scholar
  9. de Carvalho CC (2016) Whole cell biocatalysts: essential workers from Nature to the industry. Microb Biotechnol doi: 10.1111/1751-7915.12363.Google Scholar
  10. De Biase D, Pennacchietti E (2012) Glutamate decarboxylase-dependent acid resistance in orally acquired bacteria: function, distribution and biomedical implications of the gadBC operon. Mol Microbiol 86:770–786CrossRefGoogle Scholar
  11. DeBiase D, Tramonti A, John RA et al (1996) Isolation, overexpression, and biochemical characterization of the two isoforms of glutamic acid decarboxylase from Escherichia coli. Protein Expres Puri 8:430–438CrossRefGoogle Scholar
  12. Dhakal R, Bajpai VK, Baek KH (2012) Production of gaba (gamma-Aminobutyric acid) by microorganisms: a review. Braz J Microbiol 43:1230–1241CrossRefGoogle Scholar
  13. Diana M, Quilez J, Rafecas M (2014) Gamma-aminobutyric acid as a bioactive compound in foods: a review. J Funct Foods 10:407–420CrossRefGoogle Scholar
  14. Dutyshev DI et al. (2005) Structure of Escherichia coli glutamate decarboxylase (GAD alpha) in complex with glutarate at 2.05 angstrom resolution. Acta Crystal D 61:230–235CrossRefGoogle Scholar
  15. Gavrilescu M, Chisti Y (2005) Biotechnology-a sustainable alternative for chemical industry. Biotechnol Adv 23:471–499CrossRefGoogle Scholar
  16. Hao R, Schmit JC (1991) Purification and characterization of glutamate-decarboxylase from Neurospora-Crassa Conidia. J Biol Chem 266:5135–5139Google Scholar
  17. Hiraga K, Ueno Y, Oda K (2008) Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate. Biosci Biotechnol Biochem 72:1299–1306CrossRefGoogle Scholar
  18. Huang J, Le-He M, Wu H et al (2007) Biosynthesis of gamma-aminobutyric acid (GABA) using immobilized whole cells of Lactobacillus brevis. World J Microbiol Biotechnol 23:865–871CrossRefGoogle Scholar
  19. Jannoey P, Niamsup H, Lumyong S et al (2010) Comparison of gamma-aminobutyric acid production in Thai rice grains. World J Microbiol Biotechnol 26:257–263CrossRefGoogle Scholar
  20. Jeng KC, Chen CS, Fang YP et al (2007) Effect of microbial fermentation on content of statin, GABA, and polyphenols in Pu-erh tea. J Agric Food Chem 55:8787–8792CrossRefGoogle Scholar
  21. Jorge JM, Leggewie C, Wendisch VF (2016) A new metabolic route for the production of gamma-aminobutyric acid by Corynebacterium glutamicum from glucose. Amino Acids 48:2519–2531CrossRefGoogle Scholar
  22. Jun C, Joo JC, Lee JH et al (2014) Thermostabilization of glutamate decarboxylase B from Escherichia coli by structure-guided design of its pH-responsive N-terminal interdomain. J Biotechnol 174:22–28CrossRefGoogle Scholar
  23. Kanjee U, Houry WA (2013) Mechanisms of acid resistance in Escherichia coli. Annu Rev Microbiol 67:65–81CrossRefGoogle Scholar
  24. Ke CR, Yang XW, Rao HX et al (2016) Whole-cell conversion of l-glutamic acid into gamma-aminobutyric acid by metabolically engineered Escherichia coli. Springerplus 5: 591CrossRefGoogle Scholar
  25. Kim HW, Kashima Y, Ishikawa K et al (2009a) Purification and characterization of the first archaeal glutamate decarboxylase from Pyrococcus horikoshii. Biosci Biotech Bioch 73:224–227CrossRefGoogle Scholar
  26. Kim JY, Lee MY, Ji GE et al (2009b) Production of gamma-aminobutyric acid in black raspberry juice during fermentation by Lactobacillus brevis GABA100. Int J Food Microbiol 130:12–16CrossRefGoogle Scholar
  27. Kinnersley AM, Turano FJ (2000) Gamma aminobutyric acid (GABA) and plant responses to stress. Crit Rev Plant Sci 19:479–509CrossRefGoogle Scholar
  28. Komatsuzaki N, Shima J, Kawamoto S et al (2005) Production of gamma-aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods. Food Microbiol 22:497–504CrossRefGoogle Scholar
  29. Komatsuzaki N, Nakamura T, Kimura T et al (2008) Characterization of glutamate decarboxylase from a high gamma-aminobutyric acid (GABA)-producer, Lactobacillus paracasei. Biosci Biotech Bioch 72:278–285CrossRefGoogle Scholar
  30. Kook MC, Cho SC (2013) Production of GABA (gamma amino butyric acid) by lactic acid bacteria. Korean J Food Sci Anim Resour 33:377–389CrossRefGoogle Scholar
  31. Kook MC, Seo MJ, Cheigh CI, Lee SJ, Pyun YR, Park H (2010) Enhancement of gamma-amminobutyric acid production by Lactobacillus sakei B2-16 expressing glutamate decarboxylase from Lactobacillus plantarum ATCC 14917. J Korean Soc Appl Bi 53:816–820CrossRefGoogle Scholar
  32. Lee JY, Jeon SJ (2014) Characterization and immobilization on nickel-chelated Sepharose of a glutamate decarboxylase A from Lactobacillus brevis BH2 and its application for production of GABA. Biosci Biotechnol Biochem 78:1656–1661CrossRefGoogle Scholar
  33. Lee BJ, Kim JS, Kang YM et al (2010) Antioxidant activity and gamma-aminobutyric acid (GABA) content in sea tangle fermented by Lactobacillus brevis BJ20 isolated from traditional fermented foods. Food Chem 122:271–276CrossRefGoogle Scholar
  34. Lee S, Ahn J, Kim YG, Jung JK et al (2013) Gamma-aminobutyric acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography. Int J Mol Sci 14:1728–1739CrossRefGoogle Scholar
  35. Li HX, Cao YS (2010) Lactic acid bacterial cell factories for gamma-aminobutyric acid. Amino Acids 39:1107–1116CrossRefGoogle Scholar
  36. Li H, Qiu T, Gao D, Cao Y (2010a) Medium optimization for production of gamma-aminobutyric acid by Lactobacillus brevis NCL912. Amino Acids 38:1439–1445CrossRefGoogle Scholar
  37. Li H, Qiu T, Huang G, Cao Y (2010b) Production of gamma-aminobutyric acid by Lactobacillus brevis NCL912 using fed-batch fermentation. Microb Cell Fact 9:85CrossRefGoogle Scholar
  38. Lin Q (2013) Submerged fermentation of Lactobacillus rhamnosus YS9 for gamma-aminobutyric acid (GABA) production. Braz J Microbiol 44:183–187CrossRefGoogle Scholar
  39. Liu QD, Cheng HJ, Ma XQ et al (2016) Expression, characterization and mutagenesis of a novel glutamate decarboxylase from Bacillus megaterium. Biotechnol Lett 38:1107–1113CrossRefGoogle Scholar
  40. Lu XX, Xie CY, Gu ZX (2009) Optimisation of Fermentative Parameters for GABA Enrichment by Lactococcus lactis. Czech J Food Sci 27:433–442Google Scholar
  41. Lund P, Tramonti A, De Biase D (2014) Coping with low pH: molecular strategies in neutralophilic bacteria. FEMS Microbiol Rev 38:1091–1125CrossRefGoogle Scholar
  42. Nomura M, Nakajima I, Fujita Y et al (1999) Lactococcus lactis contains only one glutamate decarboxylase gene. Microbiology 145:1375–1380CrossRefGoogle Scholar
  43. Okai N, Takahashi C, Hatada K et al (2014) Disruption of pknG enhances production of gamma-aminobutyric acid by Corynebacterium glutamicum expressing glutamate decarboxylase. AMB Express 4:20CrossRefGoogle Scholar
  44. Park H, Ahn J, Lee J et al (2012) Expression, immobilization and enzymatic properties of glutamate decarboxylase fused to a cellulose-binding domain. Int J Mol Sci 13:358–368CrossRefGoogle Scholar
  45. Park SJ, Kim EY, Noh W et al (2013) Synthesis of nylon 4 from gamma-aminobutyrate (GABA) produced by recombinant Escherichia coli. Bioprocess Biosyst Eng 36:885–892CrossRefGoogle Scholar
  46. Park JY, Jeong SJ, Kim JH (2014) Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae. Biotechnol Lett 36:1791–1799CrossRefGoogle Scholar
  47. Pennacchietti E, Lammens TM, Capitani G et al (2009) Mutation of His465 alters the pH-dependent spectroscopic properties of Escherichia coli glutamate decarboxylase and broadens the range of its activity toward more alkaline pH. J Biol Chem 284:31587–31596CrossRefGoogle Scholar
  48. Pham VD, Lee SH, Park SJ et al (2015) Production of gamma-aminobutyric acid from glucose by introduction of synthetic scaffolds between isocitrate dehydrogenase, glutamate synthase and glutamate decarboxylase in recombinant Escherichia coli. J Biotechnol 207:52–57CrossRefGoogle Scholar
  49. Pham VD, Somasundaram S, Lee SH et al (2016a) Efficient production of gamma-aminobutyric acid using Escherichia coli by co-localization of glutamate synthase, glutamate decarboxylase, and GABA transporter. J Ind Microbiol Biotechnol 43:79–86CrossRefGoogle Scholar
  50. Pham VD, Somasundaram S, Park SJ et al (2016b) Co-localization of GABA shunt enzymes for the efficient production of gamma-aminobutyric acid via GABA shunt pathway in Escherichia coli. J Microbiol. Biotech 26:710–716CrossRefGoogle Scholar
  51. Plokhov AY, Gusyatiner MM, Yampolskaya TA et al (2000) Preparation of gamma-aminobutyric acid using E. coli cells with high activity of glutamate decarboxylase. Appl Biochem Biotechnol 88:257–265CrossRefGoogle Scholar
  52. Rodrigues RC, Ortiz C, Berenguer-Murcia A et al (2013) Modifying enzyme activity and selectivity by immobilization. Chem Soc Rev 42:6290–6307CrossRefGoogle Scholar
  53. Sa HD, Park JY, Jeong SJ et al (2015) Characterization of glutamate decarboxylase (GAD) from Lactobacillus sakei A156 isolated from jeot-gal. J Microbiol Biotech 25:696–703CrossRefGoogle Scholar
  54. Seo MJ, Nam YD, Lee SY et al (2013) Expression and characterization of a glutamate decarboxylase from Lactobacillus brevis 877G producing gamma-aminobutyric acid. Biosci Biotech Bioch 77:853–856CrossRefGoogle Scholar
  55. Seok JH, Park KB, Kim YH et al (2008) Production and characterization of Kimchi with enhanced levels of gamma-aminobutyric acid. Food Sci Biotechnol 17:940–946Google Scholar
  56. Servili M, Rizzello CG, Taticchi A et al (2011) Functional milk beverage fortified with phenolic compounds extracted from olive vegetation water, and fermented with functional lactic acid bacteria. Int J Food Microbiol 147:45–52CrossRefGoogle Scholar
  57. Shan Y, Man CX, Han X et al (2015) Evaluation of improved gamma-aminobutyric acid production in yogurt using Lactobacillus plantarum NDC75017. J Dairy Sci 98:2138–2149CrossRefGoogle Scholar
  58. Shelp BJ, Bown AW, McLean MD (1999) Metabolism and functions of gamma-aminobutyric acid. Trends Plant Sci 4:446–452CrossRefGoogle Scholar
  59. Shi F, Li YX (2011) Synthesis of gamma-aminobutyric acid by expressing Lactobacillus brevis-derived glutamate decarboxylase in the Corynebacterium glutamicum strain ATCC 13032. Biotechnol Lett 33:2469–2474CrossRefGoogle Scholar
  60. Shi F, Jiang J, Li Y, Xie Y (2013) Enhancement of gamma-aminobutyric acid production in recombinant Corynebacterium glutamicum by co-expressing two glutamate decarboxylase genes from Lactobacillus brevis. J Ind Microbiol Biotechnol 40:1285–1296CrossRefGoogle Scholar
  61. Shi F, Xie YL, Jiang JJ et al (2014) Directed evolution and mutagenesis of glutamate decarboxylase from Lactobacillus brevis Lb85 to broaden the range of its activity toward a near-neutral pH. Enzyme Microb Tech 61–62:35–43CrossRefGoogle Scholar
  62. Shi X, Chang C, Ma S et al (2016) Efficient bioconversion of l-glutamate to gamma-aminobutyric acid by Lactobacillus brevis resting cells. J Ind Microbiol Biotechnol. doi: 10.1007/s10295-016-1777-z Google Scholar
  63. Shin SM, Kim H, Joo Y et al (2014) Characterization of glutamate decarboxylase from Lactobacillus plantarum and its C-terminal function for the pH dependence of activity. J Agr Food Chem 62:12186–12193CrossRefGoogle Scholar
  64. Siragusa S, De Angelis M, Di Cagno R et al (2007) Synthesis of gamma-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses. Appl Environ Microb 73:7283–7290CrossRefGoogle Scholar
  65. Strigacova J, Chovanec P, Liptaj T et al (2001) Glutamate decarboxylase activity in Trichoderma viride conidia and developing mycelia. Arch Microbiol 175:32–40CrossRefGoogle Scholar
  66. Sun TS, Zhao SP, Wang HK et al (2009) ACE-inhibitory activity and gamma-aminobutyric acid content of fermented skim milk by Lactobacillus helveticus isolated from Xinjiang koumiss in China. Eur Food Res Technol 228:607–612CrossRefGoogle Scholar
  67. Takahashi C, Shirakawa J, Tsuchidate T et al (2012) Robust production of gamma-amino butyric acid using recombinant Corynebacterium glutamicum expressing glutamate decarboxylase from Escherichia coli. Enzyme Microb Tech 51:171–176CrossRefGoogle Scholar
  68. Thu Ho NA, Hou CY, Kim WH et al (2013) Expanding the active pH range of Escherichia coli glutamate decarboxylase by breaking the cooperativeness. J Biosci Bioeng 115:154–158CrossRefGoogle Scholar
  69. Tsuchiya K, Nishimura K, Iwahara M (2003) Purification and characterization of glutamate decarboxylase from Aspergillus oryzae. Food Sci Technol Res 9:283–287CrossRefGoogle Scholar
  70. Ueno H (2000) Enzymatic and structural aspects on glutamate decarboxylase. J Mol Catal B-Enzym 10:67–79CrossRefGoogle Scholar
  71. Ueno Y, Hayakawa K, Takahashi S et al (1997) Purification and characterization of glutamate decarboxylase from Lactobacillus brevis IFO 12005. Biosci Biotechnol Biochem 61:1168–1171CrossRefGoogle Scholar
  72. Wang HK, Dong C, Chen YF et al (2010) A new probiotic cheddar cheese with high ACE-inhibitory activity and gamma-aminobutyric acid content produced with Koumiss-derived Lactobacillus casei Zhang. Food Technol Biotech 48:62–70Google Scholar
  73. Wang Q, Xin YQ, Zhang F et al (2011) Enhanced gamma-aminobutyric acid-forming activity of recombinant glutamate decarboxylase (gadA) from Escherichia coli. World J Microbiol Biotechnol 27:693–700CrossRefGoogle Scholar
  74. Wang NN, Ni YL, Shi F (2015) Deletion of odhA or pyc improves production of gamma-aminobutyric acid and its precursor l-glutamate in recombinant Corynebacterium glutamicum. Biotechnol Lett 37:1473–1481CrossRefGoogle Scholar
  75. Watanabe M, Maemura K, Kanbara K et al (2002) GABA and GABA receptors in the central nervous system and other organs. Int Rev Cytol 213(213):1–47CrossRefGoogle Scholar
  76. Wendisch VF, Jorge JMP, Perez-Garcia F et al (2016) Updates on industrial production of amino acids using Corynebacterium glutamicum. World J Microbiol Biotechnol 32:105CrossRefGoogle Scholar
  77. Yang SY, Lin Q, Lu ZX et al (2008a) Characterization of a novel glutamate decarboxylase from Streptococcus salivarius ssp thermophilus Y2. J Chem Technol Biot 83:855–861CrossRefGoogle Scholar
  78. Yang SY, Lu FX, Lu ZX et al (2008b) Production of gamma-aminobutyric acid by Streptococcus salivarius subsp. thermophilus Y2 under submerged fermentation. Amino Acids 34:473–478CrossRefGoogle Scholar
  79. Yang TW, Rao ZM, Kimani BG et al (2015) Two-step production of gamma-aminobutyric acid from cassava powder using Corynebacterium glutamicum and Lactobacillus plantarum. J Ind Microbiol Biotechnol 42:1157–1165CrossRefGoogle Scholar
  80. Yao W, Wu X, Zhu J et al (2013) In vitro enzymatic conversion of gamma-aminobutyric acid immobilization of glutamate decarboxylase with bacterial cellulose membrane (BCM) and non-linear model establishment. Enzyme Microb Technol 52:258–264CrossRefGoogle Scholar
  81. Yokoyama S, Hiramatsu J, Hayakawa K (2002) Production of gamma-aminobutyric acid from alcohol distillery lees by Lactobacillus brevis IFO-12005. J Biosci Bioeng 93:95–97CrossRefGoogle Scholar
  82. Yu K, Lin L, Hu S et al (2012) C-terminal truncation of glutamate decarboxylase from Lactobacillus brevis CGMCC 1306 extends its activity toward near-neutral pH. Enzyme Microb Technol 50:263–269CrossRefGoogle Scholar
  83. Zhang C, Lu J, Chen L et al (2014a) Biosynthesis of gamma-aminobutyric acid by a recombinant Bacillus subtilis strain expressing the glutamate decarboxylase gene derived from Streptococcus salivarius ssp thermophilus Y2. Process Biochem 49:1851–1857CrossRefGoogle Scholar
  84. Zhang RZ, Yang TW, Rao ZM et al (2014b) Efficient one-step preparation of gamma-aminobutyric acid from glucose without an exogenous cofactor by the designed Corynebacterium glutamicum. Green Chem 16:4190–4197CrossRefGoogle Scholar
  85. Zhao WR, Huang J, Peng CL et al (2014) Permeabilizing Escherichia coli for whole cell biocatalyst with enhanced biotransformation ability from l-glutamate to GABA. J Mol Catal B-Enzym 107:39–46CrossRefGoogle Scholar
  86. Zhao A, Hu X, Pan L et al (2015) Isolation and characterization of a gamma-aminobutyric acid producing strain Lactobacillus buchneri WPZ001 that could efficiently utilize xylose and corncob hydrolysate. Appl Microbiol Biotechnol 99:3191–3200CrossRefGoogle Scholar

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© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinPeople’s Republic of China
  2. 2.Key Laboratory of Systems Microbial BiotechnologyChinese Academy of SciencesTianjinPeople’s Republic of China

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