Chen Ruibing, Gao Jiaoqi, Yu Wei, Chen Xianghui, Zhai Xiaoxin, Chen Yu, Zhang Lei, Zhou Yongjin J
Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Department of Pharmaceutical Botany, School of Pharmacy, Naval Medical University, Shanghai, China.
Nat Chem Biol. 2022 May;18(5):520-529. doi: 10.1038/s41589-022-01014-6. Epub 2022 Apr 28.
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 (FADH, 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) and ferulic acid (3.8 ± 0.3 g l) 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.
合成生物学的进展使微生物宿主能够以高效、具有成本竞争力且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，而辅因子在天然产物微生物生产中的作用以及辅因子工程在很大程度上被忽视了。在这里，我们系统地改造了酿酒酵母中三种辅因子（FADH、S-腺苷-L-甲硫氨酸和NADPH）的供应和循环利用，以高水平生产许多药物分子的前体酚酸咖啡酸和阿魏酸。我们开发了定制的工程策略，用于重新构建辅因子的生物合成、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）和阿魏酸（3.8±0.3 g l）的产量达到最高。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。
