Epimedium pubescens Maxim.is a well-known traditional Chinese medicinal herb with flavonol glycosides as the major pharmaceutically active compounds.UDP-glycosyltransferases(UGTs)are a group of enzymes responsible for...Epimedium pubescens Maxim.is a well-known traditional Chinese medicinal herb with flavonol glycosides as the major pharmaceutically active compounds.UDP-glycosyltransferases(UGTs)are a group of enzymes responsible for the glycosylation of flavonoid glycosides.In this study,a genome-wide analysis was performed to identify UGT family genes in E.pubescens.As a result,a total of 339 putative UGT genes were identified,which represents the largest UGT gene family known thus far,implying a significant expansion of the UGT gene family in E.pubescens.All EpUGTs were unevenly distributed across six chromosomes,and they were classified into 17 major groups.The expression profiles showed that UGT genes were differentially expressed in roots,leaves,flowers,shoots and fruits.In particular,several EpUGTs were highly induced by high light intensity,which was consistent with the accumulation level of bioactive flavonoids in E.pubescens.Six UGT79 genes that were preferentially expressed in roots or leaves were successfully expressed in E.coli,and only the recombinant EpGT60 protein was found to be active toward 8-prenylkaempferol and icaritin to produce the key bioactive compounds baohuoside II and baohuoside I.The optimal temperature,pH,k_(m) and V_(max) were determined for the recombinant EpGT60 protein.In addition,expression of recombinant EpGT60 in E.coli cell culture led to successful production of baohuoside II when fed 8-prenylkaempferol.Our study provides a foundation for further functional characterization of UGT genes in E.pubescens and provides key candidate genes for bioengineering bioactive flavonoids in E.pubescens.展开更多
基金This work was supported by grants from the CAMS Innovation Fund for Medical Sciences(CIFMS)(2021-I2M-1-031 and 2017-I2M-3-013)the National Natural Science Foundation of China(31570306 and 81473302).
文摘Epimedium pubescens Maxim.is a well-known traditional Chinese medicinal herb with flavonol glycosides as the major pharmaceutically active compounds.UDP-glycosyltransferases(UGTs)are a group of enzymes responsible for the glycosylation of flavonoid glycosides.In this study,a genome-wide analysis was performed to identify UGT family genes in E.pubescens.As a result,a total of 339 putative UGT genes were identified,which represents the largest UGT gene family known thus far,implying a significant expansion of the UGT gene family in E.pubescens.All EpUGTs were unevenly distributed across six chromosomes,and they were classified into 17 major groups.The expression profiles showed that UGT genes were differentially expressed in roots,leaves,flowers,shoots and fruits.In particular,several EpUGTs were highly induced by high light intensity,which was consistent with the accumulation level of bioactive flavonoids in E.pubescens.Six UGT79 genes that were preferentially expressed in roots or leaves were successfully expressed in E.coli,and only the recombinant EpGT60 protein was found to be active toward 8-prenylkaempferol and icaritin to produce the key bioactive compounds baohuoside II and baohuoside I.The optimal temperature,pH,k_(m) and V_(max) were determined for the recombinant EpGT60 protein.In addition,expression of recombinant EpGT60 in E.coli cell culture led to successful production of baohuoside II when fed 8-prenylkaempferol.Our study provides a foundation for further functional characterization of UGT genes in E.pubescens and provides key candidate genes for bioengineering bioactive flavonoids in E.pubescens.
