Zhang Xiaomeng, Xie Jingcong, Cao Shiping, Zhang Haiyan, Pei Jianjun, Bu Su, Zhao Linguo
Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, Nanjing, 210037, China.
College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China.
Appl Microbiol Biotechnol. 2023 May;107(9):2831-2842. doi: 10.1007/s00253-023-12464-w. Epub 2023 Mar 17.
Baicalein-7-O-glucoside and baicalein-7-O-rhamnoside have been proven to possess many pharmacological activities and are potential candidate drug leads and herb supplements. However, their further development is largely limited due to low content in host plants. Few studies reported that both bioactive plant components are prepared through the bioconversion of baicalein that is considered as the common biosynthetic precursor of both compounds. Herein, we constructed a series of the engineered whole-cell bioconversion systems in which the deletion of competitive genes and the introduction of exogenous UDP-glucose supply pathway, glucosyltransferase, rhamnosyltransferase, and the UDP-rhamnose synthesis pathway are made. Using these engineered strains, the precursor baicalein is able to be transformed into baicalein-7-O-glucoside and baicalein-7-O-rhamnoside, with high-titer production, respectively. The further optimization of fermentation conditions led to the final production of 568.8 mg/L and 877.0 mg/L for baicalein-7-O-glucoside and baicalein-7-O-rhamnoside, respectively. To the best of our knowledge, it is the highest production in preparation of baicalein-7-O-glucoside from baicalein so far, while the preparation of baicalein-7-O-rhamnoside is the first reported via bioconversion approach. Our study provides a reference for the industrial production of high-value products baicalein-7-O-glucoside and baicalein-7-O-rhamnoside using engineered E. coli. KEY POINTS: • Integrated design for improving the intracellular UDP-glucose pool • High production of rare baicalein glycosides in the engineered E. coli • Baicalein-7-O-glucoside and baicalein-7-O-rhamnoside.
黄芩苷 -7-O-葡萄糖苷和黄芩苷 -7-O-鼠李糖苷已被证明具有多种药理活性,是潜在的候选药物先导物和草药补充剂。然而,由于它们在宿主植物中的含量较低,其进一步开发受到很大限制。很少有研究报道这两种生物活性植物成分是通过黄芩素的生物转化制备的,黄芩素被认为是这两种化合物的共同生物合成前体。在此,我们构建了一系列工程化全细胞生物转化系统,其中删除了竞争基因,并引入了外源UDP-葡萄糖供应途径、葡萄糖基转移酶、鼠李糖基转移酶和UDP-鼠李糖合成途径。使用这些工程菌株,前体黄芩素能够分别转化为高产量的黄芩苷 -7-O-葡萄糖苷和黄芩苷 -7-O-鼠李糖苷。发酵条件的进一步优化导致黄芩苷 -7-O-葡萄糖苷和黄芩苷 -7-O-鼠李糖苷的最终产量分别为568.8 mg/L和877.0 mg/L。据我们所知,这是迄今为止从黄芩素制备黄芩苷 -7-O-葡萄糖苷的最高产量,而黄芩苷 -7-O-鼠李糖苷的制备是首次通过生物转化方法报道。我们的研究为利用工程化大肠杆菌工业化生产高价值产品黄芩苷 -7-O-葡萄糖苷和黄芩苷 -7-O-鼠李糖苷提供了参考。要点:• 改善细胞内UDP-葡萄糖池的综合设计 • 工程化大肠杆菌中稀有黄芩苷的高产 • 黄芩苷 -7-O-葡萄糖苷和黄芩苷 -7-O-鼠李糖苷