Comparative Activity of Six Recombinant Stilbene Synthases in Yeast for Resveratrol Production

Abstract

Resveratrol is a nutraceutical with relevant benefits to human health. This investigation reports on the generation and evaluation of six recombinant yeast lines that produce resveratrol from p-coumaric acid. The yeast lines contained a single p-coumaric acid-Co-A ligase from Plagiochasma appendiculatum combined with the stilbene synthases from Parthenocissus henryana, Polygonum cuspidatum, Morus alba var. atropurpurea, Rheum tataricum, Vitis vinifera and Arachis hypogaea. Codon optimized versions of these sequences were inserted in an expression vector flanked by the constitutive PGK and GPD promoters before expression in Saccharomyces cerevisiae. Batch fermentation (60 h) revealed that yeast lines had different capacities (p −1 p-coumaric acid improved resveratrol yields. Among the six lines, those containing the stilbene synthases (STS) from P. cuspidatum and M. alba produced up to 39 mg L−1 using 70 mg L−1 p-coumaric acid. On the other hand, lines expressing STS from V. vinifera, A. hypogaea and R. tataricum generated resveratrol faster than other lines but accumulated lower amounts at the end of the batch period (27–30 mg L−1). The simultaneous consumption of ethanol and p-coumaric acid corroborates the role of ethanol as a carbon source involved in the conversion of p-coumaric acid into resveratrol.

Keywords

• resveratrol production
• 4CL
• STS
• engineered yeasts