Yan Zhi-Bo, Liang Jing-Long, Niu Fu-Xing, Shen Yu-Ping, Liu Jian-Zhong
Institute of Synthetic Biology, Biomedical Center, Guangdong Province Key Laboratory of Improved Variety Reproduction in Aquatic Economic Animals, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
College of Light Industry and Food Science, Zhongkai University of Agriculture and Engineering, Guangzhou, China.
Front Microbiol. 2021 Oct 7;12:710405. doi: 10.3389/fmicb.2021.710405. eCollection 2021.
Pterostilbene is a derivative of resveratrol with a higher bioavailability and biological activity, which shows antioxidant, anti-inflammatory, antitumor, and antiaging activities. Here, directed evolution and host strain engineering were used to improve the production of pterostilbene in . First, the heterologous biosynthetic pathway enzymes of pterostilbene, including tyrosine ammonia lyase, -coumarate: CoA ligase, stilbene synthase, and resveratrol O-methyltransferase, were successively directly evolved through error-prone polymerase chain reaction (PCR). Four mutant enzymes with higher activities of and were obtained. The directed evolution of the pathway enzymes increased the pterostilbene production by 13.7-fold. Then, a biosensor-guided genome shuffling strategy was used to improve the availability of the precursor L-tyrosine of the host strain TYR-30 used for the production of pterostilbene. A shuffled strain with higher L-tyrosine production was obtained. The shuffled strain harboring the evolved pathway produced 80.04 ± 5.58 mg/l pterostilbene, which is about 2.3-fold the highest titer reported in literatures.
紫檀芪是白藜芦醇的衍生物，具有更高的生物利用度和生物活性，表现出抗氧化、抗炎、抗肿瘤和抗衰老活性。在此，利用定向进化和宿主菌株工程来提高紫檀芪在……中的产量。首先，通过易错聚合酶链反应（PCR）相继直接进化了紫檀芪的异源生物合成途径酶，包括酪氨酸解氨酶、对香豆酸：辅酶A连接酶、芪合酶和白藜芦醇O-甲基转移酶。获得了四种活性更高的突变酶。途径酶的定向进化使紫檀芪产量提高了13.7倍。然后，采用生物传感器引导的基因组重排策略来提高用于生产紫檀芪的宿主菌株TYR-30的前体L-酪氨酸的可用性。获得了一株L-酪氨酸产量更高的重排菌株。携带进化途径的重排菌株产生了80.04±5.58mg/l的紫檀芪，约为文献报道的最高产量的2.3倍。
