Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli

David R. Nielsen, Sang Hwal Yoon, Clara J. Yuan, Kristala L.J. Prather

Research output: Contribution to journalArticlepeer-review

87 Scopus citations

Abstract

The functional reconstruction of acetoin and meso-2,3-butanediol (meso-2,3-BD) biosynthetic pathways in Escherichia coli have been explored systematically. Pathway construction involved the in vivo screening of prospective pathway isozymes of yeast and bacterial origin. After substantial engineering of the host background to increase pyruvate availability, E. coli YYC202(DE3) ldhA- ilvC- expressing ilvBN from E. coli and aldB from L. lactis (encoding acetolactate synthase and acetolactate decarboxylase activities, respectively) was able to produce up to 870 mg/L acetoin, with no coproduction of diacetyl observed. These strains were also found to produce small quantities of meso-2,3-BD, suggesting the existence of endogenous 2,3-BD dehydrogenase activity. Finally, the coexpression of bdh1 from S. cerevisiae, encoding 2,3-BD dehydrogenase, in this strain resulted in the production of up to 1120 mg/L meso-2,3-BD, with glucose a yield of 0.29 g/g. While disruption of the native lactate biosynthesis pathway increased pyruvate precursor availability to this strain, increased availability of NADH for acetoin reduction to meso-2,3-BD was found to be the most important consequence of ldhA deletion.

Original languageEnglish (US)
Pages (from-to)274-284
Number of pages11
JournalBiotechnology Journal
Volume5
Issue number3
DOIs
StatePublished - Mar 2010
Externally publishedYes

Keywords

  • 2,3-butanediol
  • Acetoin
  • Escherichia coli
  • Metabolic engineering
  • NADH

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

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