title: Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase.
abstract: 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.
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.|Department of Science and Environment, Roskilde University, Universitetsvej 1, DK-4000, Roskilde, Denmark.
authors.forename: Beata Joanna
authors.initials: BJ
authors.lastname: Lehka
authors.affiliation: Evolva SA, Duggingerstrasse 23, CH-4153, Reinach, Switzerland.|Department of Biology, Technical University Darmstadt, Schnittspahnstrasse 10, DE-64287, Darmstadt, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Eichenberger
authors.affiliation: Department of Drug Design and Pharmacology, University of Copenhagen, Jagtvej 162, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Walden Emil
authors.initials: WE
authors.lastname: Bjørn-Yoshimoto
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.|Department of systems Biology, Technical University of Denmark, Kemitorvet Building 208, DK-2800, Kgs. Lyngby, Denmark.
authors.forename: Katherina Garcia
authors.initials: KG
authors.lastname: Vanegas
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Nicolaas
authors.initials: N
authors.lastname: Buijs
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Niels Bjerg
authors.initials: NB
authors.lastname: Jensen
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Jane Dannow
authors.initials: JD
authors.lastname: Dyekjær
authors.affiliation: Department of Science and Environment, Roskilde University, Universitetsvej 1, DK-4000, Roskilde, Denmark.
authors.forename: Håvard
authors.initials: H
authors.lastname: Jenssen
authors.affiliation: Evolva Biotech A/S, Lersø Parkallé 42, DK-2100, Copenhagen Ø, Denmark.|Evolva SA, Duggingerstrasse 23, CH-4153, Reinach, Switzerland.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Simon
authors.affiliation: Evolva SA, Duggingerstrasse 23, CH-4153, Reinach, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Naesby
mesh_terms: D020043:Genes, Essential; D006571:Heterocyclic Compounds / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D006571:Heterocyclic Compounds; D050505:Mutant Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D044925:Oxidoreductases Acting on CH-CH Group Donors; C418219:TSC13 protein, S cerevisiae
keywords: Saccharomyces cerevisiae; TSC13; flavonoids; phloretic acid