초록
<P><B>Abstract</B></P> <P>Resveratrol is a natural antioxidant compound, used as food supplement and cosmetic ingredient. Microbial production of resveratrol has until now been achieved by supplementation of expensive substrates, <I>p</I>-coumaric acid or aromatic amino acids. Here we engineered the yeast <I>Saccharomyces cerevisiae</I> to produce resveratrol directly from glucose or ethanol via tyrosine intermediate. First we introduced the biosynthetic pathway, consisting of tyrosine ammonia-lyase from <I>Herpetosiphon aurantiacus</I>, 4-coumaryl-CoA ligase from <I>Arabidopsis thaliana</I> and resveratrol synthase from <I>Vitis vinifera</I>, and obtained 2.73±0.05mgL<SUP>−1</SUP> resveratrol from glucose. Then we over-expressed feedback-insensitive alleles of <I>ARO</I>4 encoding 3-deoxy-D-arabino-heptulosonate-7-phosphate and <I>ARO</I>7 encoding chorismate mutase, resulting in production of 4.85±0.31mgL<SUP>−1</SUP> resveratrol from glucose as the sole carbon source. Next we improved the supply of the precursor malonyl-CoA by over-expressing a post-translational de-regulated version of the acetyl-CoA carboxylase encoding gene <I>ACC</I>1; this strategy further increased resveratrol production to 6.39±0.03mgL<SUP>−1</SUP>. Subsequently, we improved the strain by performing multiple-integration of pathway genes resulting in resveratrol production of 235.57±7.00mgL<SUP>−1</SUP>. Finally, fed-batch fermentation of the final strain with glucose or ethanol as carbon source resulted in a resveratrol titer of 415.65 and 531.41mgL<SUP>−1</SUP>, respectively.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Resveratrol is a polyphenolic plant secondary metabolite with antioxidant properties. </LI> <LI> Engineered <I>S. cerevisiae</I> can produce resveratrol <I>de novo</I> from glucose or ethanol. </LI> <LI> Resveratrol biosynthesis is via tyrosine ammonia lyase. </LI> <LI> 0.5gL<SUP>−1</SUP> resveratrol was produced in fed-batch fermentation on mineral medium. </LI> </UL> </P>