Engineered Production of Short-Chain Acyl-Coenzyme A Esters in Saccharomyces cerevisiae

Nicolas Krink-Koutsoubelis, Anne C. Loechner, Anna Lechner, Hannes Link, Charles M. Denby, Bastian Vögeli, Tobias J. Erb, Satoshi Yuzawa, Tadas Jakociunas, Leonard Katz, Michael K. Jensen, Victor Sourjik, Jay D Keasling*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Short-chain acyl-coenzyme A esters serve as intermediate compounds in fatty acid biosynthesis, and the production of polyketides, biopolymers and other value-added chemicals. S. cerevisiae is a model organism that has been utilized for the biosynthesis of such biologically and economically valuable compounds. However, its limited repertoire of short-chain acyl-CoAs effectively prevents its application as a production host for a plethora of natural products. Therefore, we introduced biosynthetic metabolic pathways to five different acyl-CoA esters into S. cerevisiae. Our engineered strains provide the following acyl-CoAs: propionyl-CoA, methylmalonyl-CoA, n-butyryl-CoA, isovaleryl-CoA and n-hexanoyl-CoA. We established a yeast-specific metabolite extraction protocol to determine the intracellular acyl-CoA concentrations in the engineered strains. Propionyl-CoA was produced at 4-9 μM; methylmalonyl-CoA at 0.5 μM; and isovaleryl-CoA, n-butyryl-CoA, and n-hexanoyl-CoA at 6 μM each. The acyl-CoAs produced in this study are common building blocks of secondary metabolites and will enable the engineered production of a variety of natural products in S. cerevisiae. By providing this toolbox of acyl-CoA producing strains, we have laid the foundation to explore S. cerevisiae as a heterologous production host for novel secondary metabolites.
Original languageEnglish
JournalA C S Synthetic Biology
Volume7
Issue number4
Pages (from-to)1105-1115
Number of pages11
ISSN2161-5063
DOIs
Publication statusPublished - 2018

Keywords

  • Metabolic engineering
  • S. cerevisiae
  • Heterologous pathway
  • Precursor engineering
  • Platform molecules
  • Acyl-coenzyme A

Cite this

Krink-Koutsoubelis, N., Loechner, A. C., Lechner, A., Link, H., Denby, C. M., Vögeli, B., Erb, T. J., Yuzawa, S., Jakociunas, T., Katz, L., Jensen, M. K., Sourjik, V., & Keasling, J. D. (2018). Engineered Production of Short-Chain Acyl-Coenzyme A Esters in Saccharomyces cerevisiae. A C S Synthetic Biology, 7(4), 1105-1115. https://doi.org/10.1021/acssynbio.7b00466