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Enchancement of Gamma-Aminobutyric Acid Production by Co-Localization of Neurospora crassa OR74A Glutamate Decarboxylase with Escherichia coli GABA Transporter Via Synthetic Scaffold Complex
1School of Chemical Engineering, University of Ulsan, Ulsan 44610, Republic of Korea, 2Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea, 3Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), Ulsan 44429, Republic of Korea, 3Department of Green Chemistry and Environmental Biotechnology, Korea University of Science and Technology (UST), Daejeon 34144, Republic of Korea
J. Microbiol. Biotechnol. 2017; 27(9): 1664-1669
Published September 28, 2017 https://doi.org/10.4014/jmb.1611.11041
Copyright © The Korean Society for Microbiology and Biotechnology.
Abstract
Keywords
References
- Castanie-Cornet MP, Penfound TA, Smith D, Elliott JF, Foster JW. 1999. Control of acid resistance in Escherichia coli. J. Bacteriol. 181: 3525-3535.
- Kim SH, Shin BH, Kim YH, Nam SW, Jeon SY. 2007. Cloning and expression of a full-length glutamate decarboxylase gene from Lactobacillus brevis BH2. Biotechnol. Bioprocess Eng. 12:707-712.
- Saskiawan I. 2008. Biosynthesis of polyamide 4, a biobased and biodegradable polymer. Microbiol. Indonesia 2: 119-123.
- Capitani G, De Biase D, Aurizi C, Gut H, Bossa F, Gruetter GM. 2003. Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. EMBO J. 22: 4027-4037.
- Park KB, Oh SH. 2006. Enhancement of gama-aminobutyric acid production in chungkukjang by applying a Bacillus subtilis strain expressing glutamate decarboxylase from Lactobacillus brevis. Biotechnol. Lett. 28: 1459-1463.
- Park KB, Ji GE, Park MS, Oh SH. 2005. Expression of rice glutamate decarboxylase in Bifidobacterium longum enhances γ-aminobutyric acid production. Biotechnol. Lett. 27: 1681-1684.
- Vo TD, Kim TW, Hong SH. 2012. Effects of glutamate decarboxylase and gamma-aminobutyric acid (GABA) transporter on the bioconversion of GABA in engineered Escherichia coli. Bioprocess Biosyst. Eng. 35: 645-650.
- Moon TS, Dueber JE, Shiue E, Prather KL. 2010. Use of modular, synthetic scaffold for improved production of glucaric acid in engineered E. coli. Metab. Eng. 12: 298-305.
- Hao R, Schmit JC. 1993. Cloning of the gene for glutamate decarboxylase and its expression during conidiation in Neurospora crassa. Biochem. J. 293: 735-738.
- Galagan JE, Calvo SE, Borkovich KA, Selker EU, Read ND, Jaffe D, et al. 2003. The genome sequence of the filamentous fungus Neurospora crassa. Nature 422: 859-868.
- Vo TD, Ko JS, Lee SH, Park SJ, Hong SH. 2013. Overexpression of Neurospora crassa OR74A glutamate decarboxylase in Escherichia coli for efficient GABA production. Biotechnol. Bioprocess Eng. 18: 1062-1066.
- Sambrook J, Russell DW. 2001. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
- Dueber JE, Wu GC, Malmirchegini GR, Moon TS, Petzold CJ, Ullal AV, et al. 2009. Synthetic protein scaffolds provide modular control over metabolic flux. Nat. Biotechnol. 27: 753-759.
- Baek JM, Mazumdar S, Lee SW, Jung MY, Lim JH, Seo SW, et al. 2013. Butyrate production in engineered Escherichia coli with synthetic scaffolds. Biotechnol. Bioeng. 110: 2790-2794.
- Tsai MF, McCarthy P, Miller C. 2013. Substrate selectivity in glutamate-dependent acid resistance in enteric bacteria. Proc. Natl. Acad. Sci. USA 110: 5898-5902.
- M a D, L u P, Y an C , Fan C, Y in P , Wang J, et al. 2012. Structure and mechanism of a glutamate-GABA antiporter. Nature 483: 632-636.
Related articles in JMB

Article
Research article
J. Microbiol. Biotechnol. 2017; 27(9): 1664-1669
Published online September 28, 2017 https://doi.org/10.4014/jmb.1611.11041
Copyright © The Korean Society for Microbiology and Biotechnology.
Enchancement of Gamma-Aminobutyric Acid Production by Co-Localization of Neurospora crassa OR74A Glutamate Decarboxylase with Escherichia coli GABA Transporter Via Synthetic Scaffold Complex
Sivachandiran Somasundaram 1, Murali kannan Maruthamuthu 1, Irisappan Ganesh 2, Gyeong Tae Eom 3, 4 and Soon Ho Hong 1*
1School of Chemical Engineering, University of Ulsan, Ulsan 44610, Republic of Korea, 2Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea, 3Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), Ulsan 44429, Republic of Korea, 3Department of Green Chemistry and Environmental Biotechnology, Korea University of Science and Technology (UST), Daejeon 34144, Republic of Korea
Abstract
Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant
biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate
decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving
the Neurospora crassa glutamate decarboxylase (GadB) and Escherichia coli GABA antiporter
(GadC) to improve GABA production. To construct the complex, the SH3 domain was
attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle,
and C-terminus of E. coli GadC. In the C-terminus model, 5.8 g/l of GABA concentration was
obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the
final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of
GABA yield. With the introduction of the scaffold complex, the GABA productivity increased
by 2.9 folds during the initial culture period.
Keywords: Gamma-aminobutyric acid, glutamate decarboxylase, glutamate/GABA antiporter, Neurospora crassa, synthetic protein scaffold
References
- Castanie-Cornet MP, Penfound TA, Smith D, Elliott JF, Foster JW. 1999. Control of acid resistance in Escherichia coli. J. Bacteriol. 181: 3525-3535.
- Kim SH, Shin BH, Kim YH, Nam SW, Jeon SY. 2007. Cloning and expression of a full-length glutamate decarboxylase gene from Lactobacillus brevis BH2. Biotechnol. Bioprocess Eng. 12:707-712.
- Saskiawan I. 2008. Biosynthesis of polyamide 4, a biobased and biodegradable polymer. Microbiol. Indonesia 2: 119-123.
- Capitani G, De Biase D, Aurizi C, Gut H, Bossa F, Gruetter GM. 2003. Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. EMBO J. 22: 4027-4037.
- Park KB, Oh SH. 2006. Enhancement of gama-aminobutyric acid production in chungkukjang by applying a Bacillus subtilis strain expressing glutamate decarboxylase from Lactobacillus brevis. Biotechnol. Lett. 28: 1459-1463.
- Park KB, Ji GE, Park MS, Oh SH. 2005. Expression of rice glutamate decarboxylase in Bifidobacterium longum enhances γ-aminobutyric acid production. Biotechnol. Lett. 27: 1681-1684.
- Vo TD, Kim TW, Hong SH. 2012. Effects of glutamate decarboxylase and gamma-aminobutyric acid (GABA) transporter on the bioconversion of GABA in engineered Escherichia coli. Bioprocess Biosyst. Eng. 35: 645-650.
- Moon TS, Dueber JE, Shiue E, Prather KL. 2010. Use of modular, synthetic scaffold for improved production of glucaric acid in engineered E. coli. Metab. Eng. 12: 298-305.
- Hao R, Schmit JC. 1993. Cloning of the gene for glutamate decarboxylase and its expression during conidiation in Neurospora crassa. Biochem. J. 293: 735-738.
- Galagan JE, Calvo SE, Borkovich KA, Selker EU, Read ND, Jaffe D, et al. 2003. The genome sequence of the filamentous fungus Neurospora crassa. Nature 422: 859-868.
- Vo TD, Ko JS, Lee SH, Park SJ, Hong SH. 2013. Overexpression of Neurospora crassa OR74A glutamate decarboxylase in Escherichia coli for efficient GABA production. Biotechnol. Bioprocess Eng. 18: 1062-1066.
- Sambrook J, Russell DW. 2001. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
- Dueber JE, Wu GC, Malmirchegini GR, Moon TS, Petzold CJ, Ullal AV, et al. 2009. Synthetic protein scaffolds provide modular control over metabolic flux. Nat. Biotechnol. 27: 753-759.
- Baek JM, Mazumdar S, Lee SW, Jung MY, Lim JH, Seo SW, et al. 2013. Butyrate production in engineered Escherichia coli with synthetic scaffolds. Biotechnol. Bioeng. 110: 2790-2794.
- Tsai MF, McCarthy P, Miller C. 2013. Substrate selectivity in glutamate-dependent acid resistance in enteric bacteria. Proc. Natl. Acad. Sci. USA 110: 5898-5902.
- M a D, L u P, Y an C , Fan C, Y in P , Wang J, et al. 2012. Structure and mechanism of a glutamate-GABA antiporter. Nature 483: 632-636.