Characterization of three glutamate decarboxylases from Bacillus spp. for efficient γ-aminobutyric acid production

Microbial Cell Factories - Tập 20 Số 1
Lei Sun1, Yingguo Bai1, Xiu Zhang2, Cheng Zhou2, Jie Zhang1, Xiaoyun Su1, Huiying Luo1, Bin Yao1, Yuan Wang1, Tao Tu1
1State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2Ningxia Key Laboratory for the Development and Application of Microbial Resources in Extreme Environments, North Minzu University, Yinchuan 750021, China

Tóm tắt

Abstract Background

Gamma-aminobutyric acid (GABA) is an important bio-product used in pharmaceuticals and functional foods and as a precursor of the biodegradable plastic polyamide 4. Glutamate decarboxylase (GAD) converts l-glutamate (l-Glu) into GABA via decarboxylation. Compared with other methods, develop a bioconversion platform to produce GABA is of considerable interest for industrial use.

 Results

Three GAD genes were identified from three Bacillus strains and heterologously expressed in Escherichia coli BL21 (DE3). The optimal reaction temperature and pH values for three enzymes were 40 °C and 5.0, respectively. Of the GADs, GADZ11 had the highest catalytic efficiency towards l-Glu (2.19 mM− 1 s− 1). The engineered E. coli strain that expressed GADZ11 was used as a whole-cell biocatalyst for the production of GABA. After repeated use 14 times, the cells produced GABA with an average molar conversion rate of 98.6% within 14 h.

 Conclusions

Three recombinant GADs from Bacillus strains have been conducted functional identification. The engineered E. coli strain heterologous expressing GADZ1, GADZ11, and GADZ20 could accomplish the biosynthesis of l-Glu to GABA in a buffer-free reaction at a high l-Glu concentration. The novel engineered E. coli strain has the potential to be a cost-effective biotransformation platform for the industrial production of GABA.

Từ khóa


Tài liệu tham khảo

Wong CG, Bottiglieri T, Snead OC 3rd. GABA, gamma-hydroxybutyric acid, and neurological disease. Ann Neurol. 2003;54:3–12.

Diana M, Quílez J, Rafecas M. Gamma-aminobutyric acid as a bioactive compound in foods: a review. J Funct Foods. 2014;10:407–20.

Schuller HM, Al-Wade HA, Majidi M. Gamma-aminobutyric acid, a potential tumor suppressor for small airway-derived lung adenocarcinoma. Carcinogenesis. 2008;29:1979–85.

Hagiwara H, Seki T, Ariga T. The effect of pre-germinated brown rice intake on blood glucose and PAI-1 levels in streptozotocin-induced diabetic rats. Biosci Biotechnol Biochem. 2004;68:444–7.

Padgett CL, Lalive AL, Tan KR, Terunuma M, Munoz MB, Pangalos MN, Martínez-Hernández J, Watanabe M, Moss SJ, Luján R, Lüscher C, Slesinger PA. Methamphetamine-evoked depression of GABA(B) receptor signaling in GABA neurons of the VTA. Neuron. 2012;73:978–89.

Czapinski P, Blaszczyk B, Czuczwar SJ. Mechanisms of action of antiepileptic drugs. Curr Top Med Chem. 2005;5:3–14.

Kawasaki N, Nakayama A, Yamano N, Takeda S, Kawata Y, Yamamoto N. Synthesis, thermal and mechanical properties and biodegradation of branched polyamide 4. Polymer. 2005;46:9987–93.

Yamano N, Nakayama A, Kawasaki N, Aiba YS. Mechanism and characterization of polyamide 4 degradation by Pseudomonas sp. J Polym Environ. 2008;16:141–6.

Cho HH, Jeon JW, Lee MH, Lee SH, Kwon ST. Structure and physical properties of variously drawn nylon 6-ran-nylon 4 copolymer fibers. Text Sci Eng. 2011;48:150–5.

Xu N, Wei L, Liu J. Biotechnological advances and perspectives of gamma-aminobutyric acid production. World J Microbiol Biotechnol. 2017;33:1–11.

Lee SJ, Lee HS, Lee DW. Production of γ-aminobutyric acid using immobilized glutamate decarboxylase from Lactobacillus plantarum. Korean J Microbiol Biotechnol. 2015;43:300–5.

Schüürmann J, Quehl P, Festel G, Jose J. Bacterial whole-cell biocatalysts by surface display of enzymes: toward industrial application. Appl Microbiol Biotechnol. 2014;98:8031–46.

Ke C, Wei J, Ren Y, Yang X, Chen J, Huang J. Effificient gamma-aminobutyric acid bioconversion by engineered Escherichia coli. Biotechnol Biotechnol Equip. 2018;32:566–73.

Yang X, Ke C, Zhu J, Wang Y, Zeng W, Huang J. Enhanced productivity of gamma-amino butyric acid by cascade modifications of a whole-cell biocatalyst. Appl Microbiol Biotechnol. 2018;102:3623–33.

Takahashi C, Shirakawa J, Tsuchidate T, Okai N, Hatada K, Nakayama H, Tatenoc T, Oginoa C, Kondo A. Robust production of gamma-amino butyric acid using recombinant Corynebacterium glutamicum expressing glutamate decarboxylase from Escherichia coli. Enzyme Microb Technol. 2012;51(3):171–6.

Li H, Qiu T, Huang G, Cao Y. Production of gamma-aminobutyric acid by Lactobacillus brevis NCL912 using fed-batch fermentation. Microb Cell Fact. 2010;9(1):85–5.

Choi SI, Lee JW, Park SM, Lee MY, Heo TR. Improvement of γ-aminobutyric acid (GABA) production using cell entrapment of Lactobacillus brevis GABA 057. J Microbiol Biotechnol. 2006;16(4):562–8.

Shi X, Chang C, Ma S, Cheng Y, Zhang J, Gao Q. Efficient bioconversion of L-glutamate to gamma-aminobutyric acid by Lactobacillus brevis resting cells. J Ind Microbiol Biotechnol. 2017;44(4–5):697–704.

Zhang C, Lu J, Chen L, Lu F, Lu Z. Biosynthesis of gammaaminobutyric acid by a recombinant Bacillus subtilis strain expressing the glutamate decarboxylase gene derived from Streptococcus salivarius ssp. thermophilus Y2. Process Biochem. 2014;49(11):1851–7.

Lutz G, Chavarría M, Arias ML, MataSegreda JF. Microbial degradation of palm (Elaeis guieensis) biodiesel. Rev Biol Trop. 2006;54:59–63.

Kim HW, Kashima Y, Ishikawa K, Yamano N. Purification and characterization of the first archaeal glutamate decarboxylase from Pyrococcus horikoshii. Biosci Biotechnol Biochem. 2009;73:224–7.

Lim HS, Seo DH, Cha IT, Lee H, Nam YD, Seo MJ. Expression and characterization of glutamate decarboxylase from Lactobacillus brevis HYE1 isolated from kimchi. World J Microbiol Biotechnol. 2018;34:44.

Huang J, Mei L, Sheng Q, Yao S, Lin D. Purification and characterization of glutamate decarboxylase of Lactobacillus brevis CGMCC 1306 isolated from fresh milk. Chin J Chem Eng. 2007;15:157–61.

Kim SH, Shin BH, Kim YH, Nam SW, Jeon SJ. Cloning and expression of a full-length glutamate decarboxylase gene from Lactobacillus brevisemclose BH2. Biotechnol Bioprocess Eng. 2007;12:707–12.

Ueno H. Enzymatic and structural aspects on glutamate decarboxylase. J Mol Catal B Enzym. 2000;10:67–79.

Bremer E, Krämer R. Responses of microorganisms to osmotic stress. Annu Rev Microbiol. 2019;73:313–34.

Capitani G, De Biase D, Aurizi C, Gut H, Bossa F, Grütter MG. Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. EMBO J. 2003;22:4027–37.

Global and China gamma-butyric acid (GABA) receptor market development trend report 2020–2026 industry survey and trend analysis report. Report on the status and prospects of China's γ-aminobutyric acid market (2020-2026). 2020.

Global. GABA industry survey and trend analysis report in 2021. Global Info Research. Research report on the status analysis and development trend of Global and China's γ-aminobutyric acid industry (2021-2027). 2021. Available at: https://www.cir.cn/.

Huang J, Fang H, Gai ZC, Mei JQ, Li JN, Hu S, Lv CJ, Zhao WR, Mei LH. Lactobacillus brevis CGMCC 1306 glutamate decarboxylase: Crystal structure and functional analysis. Biochem Biophys Res Co. 2018;503(3):1703–9.

Tavakoli Y, Esmaeili A, Saber H. Increasing thermal stability and catalytic activity of glutamate decarboxylase in E. coli: an in silico study. Comput Biol Chem. 2016;64:74–81.

Shin SM, Kim H, Joo Y, Lee SJ, Lee YJ, Lee SJ, Lee DW. Characterization of glutamate decarboxylase from Lactobacillus plantarum and its C-terminal function for the pH dependence of activity. J Agric Food Chem. 2014;62:12186–93.

Seo MJ, Nam YD, Lee SY, Park SL, Yi SH, Lim SI. Expression and characterization of a glutamate decarboxylase from Lactobacillus brevis 877G producing γ-aminobutyric acid. Biosci Biotechnol Biochem. 2013;77:853–6.

Komatsuzaki N, Nakamura T, Kimura T, Shima J. Characterization of glutamate decarboxylase from a high γ-aminobutyric acid (GABA)-producer Lactobacillus paracasei. Biosci Biotechnol Biochem. 2008;72:278–85.

Nomura M, Nakajima I, Fujita Y, Kobayashi M, Kimoto H, Suzuki I, Aso H. Lactococcus lactis contains only one glutamate decarboxylase gene. Microbiology. 1999;145:1375–80.

Yuan H, Wang H, Fidan O, Qin Y, Xiao G, Zhan J. Identification of new glutamate decarboxylases from Streptomyces for efficient production of γ-aminobutyric acid in engineered Escherichia coli. J Biol Eng. 2019;13:24.

Liu Q, Cheng H, Ma X, Xu N, Liu J, Ma Y. Expression, characterization and mutagenesis of a novel glutamate decarboxylase from Bacillus megaterium. Biotechnol Lett. 2016;38:1107–13.

Shi F, Xie Y, Jiang J, Wang N, Li Y, Wang X. 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 Technol. 2014;61–62:35–43.

Li Z, Zhao Y, Zhou H, Luo HB, Zhan CG. Catalytic roles of coenzyme pyridoxal-5’-phosphate (PLP) in PLP-dependent enzymes: reaction pathway for methionine-γ-lyase-catalyzed l-methionine depletion. ACS Catal. 2020;10:2198–210.

Lee S, Ahn J, Kim YG, Jung JK, Lee H, Lee E, Lee EG. Gamma-aminobutyric acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography. Int J Mol Sci. 2013;14:1728–39.

Lund P, Tramonti A, De Biase D. Coping with low pH: molecular strategies in neutralophilic bacteria. FEMS Microbiol Rev. 2014;38:1091–125.

Fan LQ, Li MW, Qiu YJ, Chen QM, Jiang SJ, Shang YJ, Zhao LM. Increasing thermal stability of glutamate decarboxylase from Escherichia coli by site-directed saturation mutagenesis and its application in GABA production. J Biotechnol. 2018;278:1–9.

Kang TJ. Ho N, Pack SP. Buffer-free production of gamma-aminobutyric acid using an engineered glutamate decarboxylase from Escherichia coli. Enzyme Microb Technol. 2013;53:200–5.

Zhang H, Yao HY, Chen F, Wang X. Purification and characterization of glutamate decarboxylase from rice germ. Food Chem. 2007;101:1670–6.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.

Syu KY, Lin CL, Huang HC, Lin JK. Determination of theanine, GABA, and other amino acids in green, oolong, black, and Pu-erh teas with dabsylation and high-performance liquid chromatography. J Agric Food Chem. 2008;56:7637–43.

Chang JY, Martin P, Bernasconi R, Braun DG. High-sensitivity amino acid analysis: measurement of amino acid neurotransmitter in mouse brain. FEBS Lett. 1981;132:117–20.

Gao Q, Duan Q, Wang D, Zhang Y, Zheng C. Separation and purification of γ-Aminobutyric acid from fermentation broth by flocculation and chromatographic methodologies. J Agric Food Chem. 2013;61:1914–9.

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Microbial Cell Factories

Tập 20 Số 1

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