Advances in synthetic biology enable microbial hosts to synthesize valuable natural products in an efficient, cost-competitive and safe manner. However, current engineering endeavors focus mainly on enzyme engineering and pathway optimization, leaving the role of cofactors in microbial production of natural products and cofactor engineering largely ignored. Here we systematically engineered the supply and recycling of three cofactors (FADH₂, S-adenosyl-l-methion and NADPH) in the yeast Saccharomyces cerevisiae, for high-level production of the phenolic acids caffeic acid and ferulic acid, the precursors of many pharmaceutical molecules. Tailored engineering strategies were developed for rewiring biosynthesis, compartmentalization and recycling of the cofactors, which enabled the highest production of caffeic acid (5.5 ± 0.2 g l⁻¹) and ferulic acid (3.8 ± 0.3 g l⁻¹) in microbial cell factories. These results demonstrate that cofactors play an essential role in driving natural product biosynthesis and the engineering strategies described here can be easily adopted for regulating the metabolism of other cofactors.

合成生物学的进步使微生物宿主能够以高效、具有成本竞争力和安全的方式合成有价值的天然产物。然而，目前的工程努力主要集中在酶工程和途径优化上，而辅因子在微生物生产天然产物和辅因子工程中的作用在很大程度上被忽略了。在这里，我们系统地设计了酵母酿酒酵母中三种辅因子（FADH ₂、S-腺苷-L-甲硫氨酸和 NADPH）的供应和回收。，用于高水平生产酚酸咖啡酸和阿魏酸，这是许多药物分子的前体。制定了量身定制的工程策略，用于重新连接辅因子的生物合成、区室化和循环利用，这使得微生物细胞工厂中咖啡酸（5.5 ± 0.2 g l ^-1）和阿魏酸（3.8 ± 0.3 g l ^{-1 ）的产量最高。}这些结果表明，辅因子在驱动天然产物生物合成中起着至关重要的作用，并且这里描述的工程策略可以很容易地用于调节其他辅因子的代谢。