posted on 2018-03-02, 00:00authored byNicolas 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
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.