Anhui Engineering Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources, Department of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, 237012, Anhui, People's Republic of China.
Biotechnol Lett. 2022 Sep;44(9):1073-1080. doi: 10.1007/s10529-022-03285-4. Epub 2022 Aug 3.
To produce high concentrations of hyperoside from quercetin using recombinant Escherichia coli with in situ regeneration of UDP-galactose.
Sucrose synthase from Glycine max (GmSUS) was co-expressed with UDP-glucose epimerase from E. coli (GalE) in E. coli for regenerating UDP-galactose from UDP and sucrose. Glycosyltransferase from Petunia hybrida (PhUGT) was introduced to synthesize hyperoside from quercetin through the regeneration system of UDP-galactose. Co-expressing with molecular chaperones GroEL/ES successfully enhanced the catalytic efficiency of the recombinant strain, which assisted the soluble expression of PhUGT. By using a fed-batch approach, the production of hyperoside reached 863.7 mg L with a corresponding molar conversion of 93.6% and a specific productivity of 72.5 mg L h.
The method described herein for hyperoside production can be widely applied for the synthesis of isorhamnetin-3-O-galactoside, kaempferol-3-O-galactoside and other flavonoids.
利用重组大肠杆菌原位再生 UDP-半乳糖，从槲皮素生产高浓度的圣草酚。
在大肠杆菌中，共表达来自大豆的蔗糖合酶（GmSUS）和来自大肠杆菌的 UDP-葡萄糖差向异构酶（GalE），以从 UDP 和蔗糖中再生 UDP-半乳糖。从矮牵牛中引入糖基转移酶（PhUGT），通过 UDP-半乳糖再生系统，将槲皮素合成圣草酚。共表达分子伴侣 GroEL/ES 成功提高了重组菌株的催化效率，辅助 PhUGT 的可溶性表达。通过分批补料的方法，圣草酚的产量达到 863.7mg/L，相应的摩尔转化率为 93.6%，比生产率为 72.5mg/L·h。
本文描述的生产圣草酚的方法可广泛应用于异鼠李素-3-O-半乳糖苷、山柰酚-3-O-半乳糖苷和其他类黄酮的合成。
