Caffeic acid (CA)-derived phenethyl ester (CAPE) and phenethyl amide (CAPA) are extensively investigated bioactive compounds with therapeutic applications such as antioxidant, anti-inflammatory, and anticarcinogenic properties. To construct microbial cell factories for production of CAPE or CAPA is a promising option given the limitation of natural sources for product extraction and the environmental toxicity of the agents used in chemical synthesis. We reported the successful biosynthesis of caffeic acid in yeast previously. Here in this work, we further constructed the downstream synthetic pathways in yeast for biosynthesis of CAPE and CAPA. After combinatorial engineering of yeast chassis based on the rational pathway engineering method and library-based SCRaMbLE method, we finally obtained the optimal strains that respectively produced 417 μg/L CAPE and 1081 μg/L CAPA. Two screened gene targets of ΔHAM1 and ΔYJL028W were discovered to help improve the product synthesis capacity. This is the first report of the de novo synthesis of CAPA from glucose in an engineered yeast chassis. Future work on enzyme and chassis engineering will further support improving the microbial cell factories for the production of CA derivatives.

中文翻译：

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用于生物合成咖啡酸苯乙酯和苯乙酰胺的酵母代谢工程


咖啡酸 (CA) 衍生的苯乙酯 (CAPE) 和苯乙酰胺 (CAPA) 是经过广泛研究的生物活性化合物，具有抗氧化、抗炎和抗癌等治疗应用。考虑到产品提取天然来源的限制以及化学合成中使用的试剂的环境毒性，构建用于生产 CAPE 或 CAPA 的微生物细胞工厂是一个有前途的选择。我们之前报道过在酵母中成功生物合成咖啡酸。在这项工作中，我们进一步构建了酵母中 CAPE 和 CAPA 生物合成的下游合成途径。经过基于合理途径工程方法和基于文库的 SCRaMbLE 方法对酵母底盘进行组合工程，最终获得了分别产生 417 μg/L CAPE 和 1081 μg/L CAPA 的最佳菌株。发现了两个筛选的基因靶点ΔHAM1和ΔYJL028W，有助于提高产品合成能力。这是在工程酵母底盘中从葡萄糖从头合成 CAPA 的第一份报告。酶和底盘工程的未来工作将进一步支持改进用于生产 CA 衍生物的微生物细胞工厂。