Study on the Synthesis of Resveratrol by Recombinant Saccharomyces cerevisiae
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Abstract:
In order to obtain a system for rapid synthesis of resveratrol by Saccharomyces cerevisiae, the key enzyme genes of the resveratrol synthesis pathway were cloned in this study, stilbene synthase gene (STS) from Polygonum cuspidatum and 4-coumaroyl-CoA ligase (4CL) from Nicotiana tabacum, to construct the fusion gene of 4CL-(GSG)3-STS by overlap PCR amplification technology through troducing three consecutive repeats of GSG as a linker peptide. The recombinant expression vector pESC-TRP-4CL-(GSG)3-STS obtained was then transformed into S. cerevisiae. The metabolites of the recombinant S. cerevisiae were analyzed by HPLC. Finally, the induction time, and substrate concentration and addition method for the synthesis of resveratrol by the recombinant S. cerevisiae were further optimized. The result showed that the yield of resveratrol could reach 7.01 mg/L 8 h after the established recombinant S. cerevisiae cells, which had grown for 48 h, were induced to express while the substrate 4-coumarate was added at 6 mg/L once every 2 h. The synthesis of resveratrol using this current system is simple for operation and requires only a short production cycle, thus, provides a basis for realizing a large-scale microbial production of resveratrol.
