Lotus(Nelumbo nucifera),an ancient aquatic plant,possesses a unique pharmacological activity that is primarily contributed by benzylisoquinoline alkaloids(BIAs).However,only few genes and enzymes involved in BIA biosy...Lotus(Nelumbo nucifera),an ancient aquatic plant,possesses a unique pharmacological activity that is primarily contributed by benzylisoquinoline alkaloids(BIAs).However,only few genes and enzymes involved in BIA biosynthesis in N.nucifera have been isolated and characterized.In the present study we identified the regiopromiscuity of an O-methyltransferase,designated NnOMT6,isolated from N.nucifera;NnOMT6 was found to catalyze the methylation of monobenzylisoquinoline 6-O/7-O,aporphine skeleton 6-O,phenylpropanoid 3-O,and protoberberine 2-O.We further probed the key residues affecting NnOMT6 activity via molecular docking and molecular dynamics simulation.Verification using site-directed mutagenesis revealed that residues D316,N130,L135,N176A,D269,and E328 were critical for BIA O-methyltransferase activities;furthermore,N323A,a mutant of NnOMT6,demonstrated a substantial increase in catalytic efficiency for BIAs and a broader acceptor scope compared with wild-type NnOMT6.To the best of our knowledge,this is the first study to report the O-methyltransferase activity of an aporphine skeleton without benzyl moiety substitutions in N.nucifera.The study findings provide biocatalysts for the semisynthesis of related medical compounds and give insights into protein engineering to strengthen O-methyltransferase activity in plants.展开更多
The Asteraceae (Compositae),a large plant family of approximately 24 000-35 000 species,accounts for^10% of all angiosperm species and contributes a lot to plant diversity.The most representative members of the Astera...The Asteraceae (Compositae),a large plant family of approximately 24 000-35 000 species,accounts for^10% of all angiosperm species and contributes a lot to plant diversity.The most representative members of the Asteraceae are the economically important chrysanthemums (Chrysanthemum L.)that diversified through reticulate evolution.Biodiversity is typically created by multiple evolutionary mechanisms such as wholegenome duplication 0NGD)or polyploidization and locally repetitive genome expansion.However,the lack of genomic data from chrysanthemum species has prevented an in-depth analysis of the evolutionary mechanisms involved in their diversification.Here,we used Oxford Nanopore long-read technologyto sequence the diploid Chrysanthemum nankingense genome,which represents one of the progenitor genomes of domesticated chrysanthemums.Our analysis revealed that the evolution of the C.nankingense genome was driven by bursts of repetitive element expansion and WGD events including a recentWGD that distinguishes chrysanthemum from sunflower,which diverged from chrysanthemum approximately 38.8 million years ago.Variations of ornamental and medicinal traits in chrysanthemums are linked to the expansion of candidate gene families by duplication events including paralogous gene duplication.Collectively,our study of the assembled reference genome offers new knowledge and resources to dissect the history and pattern of evolution and diversification of chrysanthemum plants,and also to accelerate their breeding and improvement.展开更多
基金supported by the National Natural Science Foundation of China(32170388)the Scientific and Technological Innovation project of China Academy of Chinese Medical Sciences(CACMS Innovation Fund CI2021A04108,CI2021A04515)the Fundamental Research Funds for the Central Public Welfare Research Institutes of China(ZZ13-YQ-057,ZXKT21006)。
文摘Lotus(Nelumbo nucifera),an ancient aquatic plant,possesses a unique pharmacological activity that is primarily contributed by benzylisoquinoline alkaloids(BIAs).However,only few genes and enzymes involved in BIA biosynthesis in N.nucifera have been isolated and characterized.In the present study we identified the regiopromiscuity of an O-methyltransferase,designated NnOMT6,isolated from N.nucifera;NnOMT6 was found to catalyze the methylation of monobenzylisoquinoline 6-O/7-O,aporphine skeleton 6-O,phenylpropanoid 3-O,and protoberberine 2-O.We further probed the key residues affecting NnOMT6 activity via molecular docking and molecular dynamics simulation.Verification using site-directed mutagenesis revealed that residues D316,N130,L135,N176A,D269,and E328 were critical for BIA O-methyltransferase activities;furthermore,N323A,a mutant of NnOMT6,demonstrated a substantial increase in catalytic efficiency for BIAs and a broader acceptor scope compared with wild-type NnOMT6.To the best of our knowledge,this is the first study to report the O-methyltransferase activity of an aporphine skeleton without benzyl moiety substitutions in N.nucifera.The study findings provide biocatalysts for the semisynthesis of related medical compounds and give insights into protein engineering to strengthen O-methyltransferase activity in plants.
基金The National Key Research and Development Program of China (2016YFD0801102)and the National Natural Science Foundation of China (31870198 and 31400278).
文摘The Asteraceae (Compositae),a large plant family of approximately 24 000-35 000 species,accounts for^10% of all angiosperm species and contributes a lot to plant diversity.The most representative members of the Asteraceae are the economically important chrysanthemums (Chrysanthemum L.)that diversified through reticulate evolution.Biodiversity is typically created by multiple evolutionary mechanisms such as wholegenome duplication 0NGD)or polyploidization and locally repetitive genome expansion.However,the lack of genomic data from chrysanthemum species has prevented an in-depth analysis of the evolutionary mechanisms involved in their diversification.Here,we used Oxford Nanopore long-read technologyto sequence the diploid Chrysanthemum nankingense genome,which represents one of the progenitor genomes of domesticated chrysanthemums.Our analysis revealed that the evolution of the C.nankingense genome was driven by bursts of repetitive element expansion and WGD events including a recentWGD that distinguishes chrysanthemum from sunflower,which diverged from chrysanthemum approximately 38.8 million years ago.Variations of ornamental and medicinal traits in chrysanthemums are linked to the expansion of candidate gene families by duplication events including paralogous gene duplication.Collectively,our study of the assembled reference genome offers new knowledge and resources to dissect the history and pattern of evolution and diversification of chrysanthemum plants,and also to accelerate their breeding and improvement.