increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the ...increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the Artemisia annua L. plant. In this study, amorpha-4, 11-diene synthase （ADS） cDNA （ads1） and genomics gene （gads1） were cloned from a high-yield A. annua strain 001. The activity of ADS1 was confirmed by heterogeneous overexpression of ads I and in vitro enzymatic incubation. Reverse transcript-polymerase chain reaction results demonstrated that ads1 expressed in leaves, flowers and young stems, but not in roots. This organ-specific expression pattern of ads1 is consistent with that of artemisinin accumulation in the plant. The gads1 has a complex organization including seven exons and six introns, and belongs to class III terpene synthase. DNA gel blotting revealed that the ADS gene has at least four copies in the genome of strain 001. The higher copy numbers might be one of the reasons for its high artemisinin content.展开更多
Amorpha-4,11-diene synthase(ADS)catalyzes the first committed step in the artemisinin biosynthetic pathway,which is the first catalytic reaction enzymatically and genetically characterized in artemisinin biosynthesis....Amorpha-4,11-diene synthase(ADS)catalyzes the first committed step in the artemisinin biosynthetic pathway,which is the first catalytic reaction enzymatically and genetically characterized in artemisinin biosynthesis.The advent of ADS in Artemisia annua is considered crucial for the emergence of the specialized artemisinin biosynthetic pathway in the species.Microbial production of amorpha-4,11-diene is a breakthrough in metabolic engineering and synthetic biology.Recently,numerous new techniques have been used in ADS engineering;for example,assessing the substrate promiscuity of ADS to chemoenzymatically produce artemisinin.In this review,we discuss the discovery and catalytic mechanism of ADS,its application in metabolic engineering and synthetic biology,as well as the role of sesquiterpene synthases in the evolutionary origin of artemisinin.展开更多
基金Supported by the National Natural Science Foundation of China （30171740 and 30672623）.
文摘increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the Artemisia annua L. plant. In this study, amorpha-4, 11-diene synthase （ADS） cDNA （ads1） and genomics gene （gads1） were cloned from a high-yield A. annua strain 001. The activity of ADS1 was confirmed by heterogeneous overexpression of ads I and in vitro enzymatic incubation. Reverse transcript-polymerase chain reaction results demonstrated that ads1 expressed in leaves, flowers and young stems, but not in roots. This organ-specific expression pattern of ads1 is consistent with that of artemisinin accumulation in the plant. The gads1 has a complex organization including seven exons and six introns, and belongs to class III terpene synthase. DNA gel blotting revealed that the ADS gene has at least four copies in the genome of strain 001. The higher copy numbers might be one of the reasons for its high artemisinin content.
基金the National Natural Science Foundation of China(31872666)the Special Fund for Talent Introduction of Kunming Institute of Botany,CAS+2 种基金the China Postdoctoral Science Foundation(Grant Nos.2020M671252 and 2020T130668)the Young Elite Scien tists Sponsorship Program by CAST(2019QNRC001)the Open Fund of Shanghai Key Laboratory of Plant Functional Genomics and Resources(PFGR201902).
文摘Amorpha-4,11-diene synthase(ADS)catalyzes the first committed step in the artemisinin biosynthetic pathway,which is the first catalytic reaction enzymatically and genetically characterized in artemisinin biosynthesis.The advent of ADS in Artemisia annua is considered crucial for the emergence of the specialized artemisinin biosynthetic pathway in the species.Microbial production of amorpha-4,11-diene is a breakthrough in metabolic engineering and synthetic biology.Recently,numerous new techniques have been used in ADS engineering;for example,assessing the substrate promiscuity of ADS to chemoenzymatically produce artemisinin.In this review,we discuss the discovery and catalytic mechanism of ADS,its application in metabolic engineering and synthetic biology,as well as the role of sesquiterpene synthases in the evolutionary origin of artemisinin.
