TY - JOUR

T1 - Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase

AU - Lehka, Beata Joanna

AU - Eichenberger, Michael

AU - Bjørn-Yoshimoto, Walden Emil

AU - Vanegas, Katherina Garcia

AU - Buijs, Nicolaas

AU - Jensen, Niels Bjerg

AU - Dyekjær, Jane Dannow

AU - Jenssen, Håvard

AU - Simon, Ernesto

AU - Naesby, Michael

N1 - © FEMS 2017.

PY - 2017/1

Y1 - 2017/1

N2 - Phenylpropanoids, such as flavonoids and stilbenoids, are of great commercial interest, and their production in S. cerevisiae is a very promising strategy. However, to achieve commercially viable production, each step of the process must be optimized. 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.

AB - Phenylpropanoids, such as flavonoids and stilbenoids, are of great commercial interest, and their production in S. cerevisiae is a very promising strategy. However, to achieve commercially viable production, each step of the process must be optimized. 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.

U2 - 10.1093/femsyr/fox004

DO - 10.1093/femsyr/fox004

M3 - Journal article

C2 - 28073929

VL - 17

JO - FEMS Yeast Research

JF - FEMS Yeast Research

SN - 1567-1356

IS - 1

M1 - fox004

ER -