Metabolomics-driven approach to solving a CoA imbalance for improved 1-butanol production in Escherichia coli

Metab Eng. 2017 May;41:135-143. doi: 10.1016/j.ymben.2017.04.003. Epub 2017 Apr 8.


High titer 1-butanol production in Escherichia coli has previously been achieved by overexpression of a modified clostridial 1-butanol production pathway and subsequent deletion of native fermentation pathways. This strategy couples growth with production as 1-butanol pathway offers the only available terminal electron acceptors required for growth in anaerobic conditions. With further inclusion of other well-established metabolic engineering principles, a titer of 15g/L has been obtained. In achieving this titer, many currently existing strategies have been exhausted, and 1-butanol toxicity level has been surpassed. Therefore, continued engineering of the host strain for increased production requires implementation of alternative strategies that seek to identify non-obvious targets for improvement. In this study, a metabolomics-driven approach was used to reveal a CoA imbalance resulting from a pta deletion that caused undesirable accumulation of pyruvate, butanoate, and other CoA-derived compounds. Using metabolomics, the reduction of butanoyl-CoA to butanal catalyzed by alcohol dehydrogenase AdhE2 was determined as a rate-limiting step. Fine-tuning of this activity and subsequent release of free CoA restored the CoA balance that resulted in a titer of 18.3g/L upon improvement of total free CoA levels using cysteine supplementation. By enhancing AdhE2 activity, carbon flux was directed towards 1-butanol production and undesirable accumulation of pyruvate and butanoate was diminished. This study represents the initial report describing the improvement of 1-butanol production in E. coli by resolving CoA imbalance, which was based on metabolome analysis and rational metabolic engineering strategies.

Keywords: 1-Butanol; CoA balance; Escherichia coli; Metabolomics; Strain improvement.

MeSH terms

  • 1-Butanol / metabolism*
  • Alcohol Dehydrogenase* / genetics
  • Alcohol Dehydrogenase* / metabolism
  • Aldehyde Oxidoreductases* / genetics
  • Aldehyde Oxidoreductases* / metabolism
  • Coenzyme A* / genetics
  • Coenzyme A* / metabolism
  • Cysteine / metabolism
  • Escherichia coli Proteins* / genetics
  • Escherichia coli Proteins* / metabolism
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Metabolome*
  • Metabolomics*


  • Escherichia coli Proteins
  • 1-Butanol
  • Alcohol Dehydrogenase
  • adhE protein, E coli
  • Aldehyde Oxidoreductases
  • Cysteine
  • Coenzyme A