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白藜芦醇是一种极具药用价值的植物源芪类化合物。为了在E.coli实现白藜芦醇的从头合成,构建了由酪氨酸解氨酶(TAL),香豆酸-CoA合成酶(4CL)和白藜芦醇合成酶(STS)组成的非天然合成途径。经3天发酵后,白藜芦醇产量仅为2.67 mg/L,而其中间体香豆酸的积累达到了95.64 mg/L。为了进一步改善异源途径的效率,对4CL和STS模块采取融合表达、高拷贝表达及启动子工程改造的策略,最终使白藜芦醇产量提高到了9.6倍,达到了25.76 mg/L,同时香豆酸的积累减少到了20.38 mg/L。这些研究结果为更高效白藜芦醇从头合成工程菌的构建及最终实现白藜芦醇的微生物大规模生产奠定了基础。
Resveratrol is a highly medicinal plant-derived stilbene compound. In order to realize the de novo synthesis of resveratrol in E.coli, we constructed a non-transformant system consisting of tyrosine ammonia lyase (TAL), coumaric acid-CoA synthetase (4CL) and resveratrol synthase (STS) Natural synthesis pathway. After 3 days of fermentation, the yield of resveratrol was only 2.67 mg / L, while the accumulation of intermediate coumaric acid reached 95.64 mg / L. In order to further improve the efficiency of the heterologous pathway, the strategies of fusion expression, high copy expression and promoter engineering were adopted for 4CL and STS modules, which eventually increased the production of resveratrol to 9.6 times, reaching 25.76 mg / L, The accumulation of levulinic acid was reduced to 20.38 mg / L. These findings lay the foundation for a more efficient resveratrol de novo synthesis engineered bacteria and the ultimate realization of resveratrol microbial mass production.