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. 2015 Aug 10:207:52-7. doi: 10.1016/j.jbiotec.2015.04.028. Epub 2015 May 18.

Abstract

Escherichia coli were engineered for the direct production of gamma-aminobutyric acid from glucose by introduction of synthetic protein scaffold. In this study, three enzymes consisting GABA pathway (isocitrate dehydrogenase, glutamate synthase and glutamate decarboxylase) were connected via synthetic protein scaffold. By introduction of scaffold, 0.92g/L of GABA was produced from 10g/L of glucose while no GABA was produced in wild type E. coli. The optimum pH and temperature for GABA production were 4.5 and 30°C, respectively. When competing metabolic network was inactivated by knockout mutation, maximum GABA concentration of 1.3g/L was obtained from 10g/L glucose. The recombinant E. coli strain which produces GABA directly from glucose was successfully constructed by introduction of protein scaffold.

Keywords: GABA, Escherichia coli, Synthetic scaffold protein, Synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Glucose / metabolism
  • Glutamate Decarboxylase / genetics
  • Glutamate Decarboxylase / metabolism
  • Glutamate Synthase / genetics
  • Glutamate Synthase / metabolism
  • Isocitrate Dehydrogenase / genetics
  • Isocitrate Dehydrogenase / metabolism
  • Protein Engineering / methods*
  • Synthetic Biology
  • gamma-Aminobutyric Acid / biosynthesis*

Substances

  • gamma-Aminobutyric Acid
  • Isocitrate Dehydrogenase
  • Glutamate Synthase
  • Glutamate Decarboxylase
  • Glucose