Abstract
Objectives
To direct the carbon flux from Krebs cycle into the gamma-aminobutyric acid (GABA) shunt pathway for the production of GABA by protein scaffold introduction in Escherichia coli.
Results
Escherichia coli was engineered to produce GABA from glucose by the co-localization of enzymes succinate semialdehyde dehydrogenase (GadD), GABA aminotransferase (PuuE) and GABA transporter (GadC) by protein scaffold. 0.7 g GABA l−1 was produced from 10 g glucose l−1 while no GABA was produced in wild type E. coli. pH 6 and 30 °C were optimum for GABA production, and GABA concentration increased to 1.12 g GABA l−1 when 20 g glucose l−1 was used. When competing metabolic networks were inactivated, GABA increased by 24 % (0.87 g GABA l−1).
Conclusions
The novel GABA production system was constructed by co-localization of GABA shunt enzymes.
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Acknowledgments
This work was supported by a grant from the Next-Generation BioGreen 21 Program (SSAC, Grant Number: PJ01111601), Rural Development Administration, Republic of Korea.
Supporting information
Supplementary Table 1—List of bacterial strains and plasmids used in this study.
Supplementary Table 2—Primers used in this work.
Supplementary Fig. 1—a Scaffold device system pBCDE plasmids were used in this study. b Results of the SDS-PAGE analysis of the overexpressed three genes.
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Pham, V.D., Somasundaram, S., Lee, S.H. et al. Engineering the intracellular metabolism of Escherichia coli to produce gamma-aminobutyric acid by co-localization of GABA shunt enzymes. Biotechnol Lett 38, 321–327 (2016). https://doi.org/10.1007/s10529-015-1982-2
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DOI: https://doi.org/10.1007/s10529-015-1982-2