Journal article
Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase
Roskilde University1
Technical University of Darmstadt2
University of Copenhagen3
Department of Biotechnology and Biomedicine, Technical University of Denmark4
Eukaryotic Molecular Cell Biology, Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark5
Evolva Biotech A/S6
Evolva SA7
Phenylpropanoids, such as flavonoids and stilbenoids, are of great commercial interest, and their production in Saccharomyces cerevisiae is a very promising strategy. However, to achieve commercially viable production, each step of the process must be optimised. We looked at carbon loss, known to occur in the heterologous flavonoid pathway in yeast, and identified an endogenous enzyme, the enoyl reductase Tsc13, which turned out to be responsible for the accumulation of phloretic acid via reduction of p-coumaroyl-CoA.
Tsc13 is an essential enzyme involved in fatty acid synthesis and cannot be deleted. Hence, two approaches were adopted in an attempt to reduce the side activity without disrupting the natural function: site saturation mutagenesis identified a number of amino acid changes which slightly increased flavonoid production but without reducing the formation of the side product.
Conversely, the complementation of TSC13 by a plant gene homologue essentially eliminated the unwanted side reaction, while retaining the productivity of phenylpropanoids in a simulated fed batch fermentation.
Language: | English |
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Publisher: | Oxford University Press |
Year: | 2016 |
Pages: | fox004 |
ISSN: | 15671364 and 15671356 |
Types: | Journal article |
DOI: | 10.1093/femsyr/fox004 |
ORCIDs: | Garcia Vanegas, Katherina and 0000-0003-1019-9239 |