biostudies-literatureWang ZLNational Natural Science Foundation of ChinaNational Postdoctoral Program for Innovative Talents4418-4425https://www.ebi.ac.uk/biostudies/studies/S-EPMC10132108biostudies-literatureUnknown14(16)A highly efficient and promiscuous 7,4'-di-O-glycosyltransferase ZjOGT3 was discovered from the medicinal plant Ziziphus jujuba var. spinosa. ZjOGT3 could sequentially catalyse 4'- and 7-O-glycosylation of flavones to produce 7,4'-di-O-glycosides with obvious regio-selectivity. For 7,4'-dihydroxyl flavanones and 3-O-glycosylated 7,4'-dihydroxyl flavones, ZjOGT3 selectively catalyses 7-O-glycosylation. The crystal structure of ZjOGT3 was solved. Structural analysis, DFT calculations, MD simulations, and site-directed mutagenesis reveal that the regio-selectivity is mainly controlled by the enzyme microenvironment for 7,4'-dihydroxyl flavones and 3-O-glycosylated 7,4'-dihydroxyl flavones. For 7,4'-dihydroxyl flavanones, the selectivity Chemical scienceFunctional characterization, structural basis, and regio-selectivity control of a promiscuous flavonoid 7,4'-di-O-glycosyltransferase from Ziziphus jujuba var. spinosa.PMC10132108BX2022002281725023Ye MQiao XChen KZhou JJLiang YWang HDWang HTWei WZhou HWang ZLLi FDWang RSfalseFunctional characterization, structural basis, and regio-selectivity control of a promiscuous flavonoid 7,4'-di-O-glycosyltransferase from Ziziphus jujuba var. spinosa.A highly efficient and promiscuous 7,4'-di-O-glycosyltransferase ZjOGT3 was discovered from the medicinal plant Ziziphus jujuba var. spinosa. ZjOGT3 could sequentially catalyse 4'- and 7-O-glycosylation of flavones to produce 7,4'-di-O-glycosides with obvious regio-selectivity. For 7,4'-dihydroxyl flavanones and 3-O-glycosylated 7,4'-dihydroxyl flavones, ZjOGT3 selectively catalyses 7-O-glycosylation. The crystal structure of ZjOGT3 was solved. Structural analysis, DFT calculations, MD simulations, and site-directed mutagenesis reveal that the regio-selectivity is mainly controlled by the enzyme microenvironment for 7,4'-dihydroxyl flavones and 3-O-glycosylated 7,4'-dihydroxyl flavones. For 7,4'-dihydroxyl flavanones, the selectivity 2023-01-01T00:00:00Z2023 Apr2026-05-29T06:37:27.456Z2024-11-13T07:31:38.29ZS-EPMC101321083712317710.1039/d2sc06504e