Amorpha-4,11-diene synthase: a key enzyme in artemisinin biosynthesis and engineering

Huang, Jin; Fang, Xin

doi:10.1007/s42994-021-00058-x

Cited by 5 publications

(3 citation statements)

References 97 publications

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“…A strong positive association was found between artemisinin levels and the number of copies of AaADS [71]. ADS catalyzes the initial and critical step in the artemisinin biosynthetic pathway, the conversion of farnesyl diphosphate (FDP) to amorpha-4, 11-diene [90]. Overexpression of AaADS can enhance artemisinin production in transgenic A. annua plants, suggesting the important role of AaADS in artemisinin biosynthesis [91,92].…”

Section: Terpenoid Biosynthesis Pathway and Regulationmentioning

confidence: 96%

Functions of Representative Terpenoids and Their Biosynthesis Mechanisms in Medicinal Plants

Wang,

Zhao,

Jiang

et al. 2023

Biomolecules

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Terpenoids are the broadest and richest group of chemicals obtained from plants. These plant-derived terpenoids have been extensively utilized in various industries, including food and pharmaceuticals. Several specific terpenoids have been identified and isolated from medicinal plants, emphasizing the diversity of biosynthesis and specific functionality of terpenoids. With advances in the technology of sequencing, the genomes of certain important medicinal plants have been assembled. This has improved our knowledge of the biosynthesis and regulatory molecular functions of terpenoids with medicinal functions. In this review, we introduce several notable medicinal plants that produce distinct terpenoids (e.g., Cannabis sativa, Artemisia annua, Salvia miltiorrhiza, Ginkgo biloba, and Taxus media). We summarize the specialized roles of these terpenoids in plant-environment interactions as well as their significance in the pharmaceutical and food industries. Additionally, we highlight recent findings in the fields of molecular regulation mechanisms involved in these distinct terpenoids biosynthesis, and propose future opportunities in terpenoid research, including biology seeding, and genetic engineering in medicinal plants.

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Section: Terpenoid Biosynthesis Pathway and Regulationmentioning

confidence: 96%

Functions of Representative Terpenoids and Their Biosynthesis Mechanisms in Medicinal Plants

Wang,

Zhao,

Jiang

et al. 2023

Biomolecules

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“…The biosynthetic pathways of artemisinin begin with amorpha-4,11-diene synthase (ADS), which converts farnesyl diphosphate (FPP) into the amorphadiene skeleton ( Bouwmeester et al., 1999 ; Chang et al., 2000 ; Mercke et al., 2000 ; Huang and Fang, 2021 ). Subsequently, a cytochrome P450-dependent monooxygenase known as CYP71AV1 performs a three-step oxidation of amorpha-4,11-diene, resulting in the generation of minor products of artemisinic alcohol and artemisinic aldehyde and a major product of artemisinic acid in yeast ( Ro et al., 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Functional analysis of CYP71AV1 reveals the evolutionary landscape of artemisinin biosynthesis

Chen,

Mu,

et al. 2024

Front. Plant Sci.

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Artemisinin biosynthesis, unique to Artemisia annua, is suggested to have evolved from the ancestral costunolide biosynthetic pathway commonly found in the Asteraceae family. However, the evolutionary landscape of this process is not fully understood. The first oxidase in artemisinin biosynthesis, CYP71AV1, also known as amorpha-4,11-diene oxidase (AMO), has specialized from ancestral germacrene A oxidases (GAOs). Unlike GAO, which exhibits catalytic promiscuity toward amorpha-4,11-diene, the natural substrate of AMO, AMO has lost its ancestral activity on germacrene A. Previous studies have suggested that the loss of the GAO copy in A. annua is responsible for the abolishment of the costunolide pathway. In the genome of A. annua, there are two copies of AMO, each of which has been reported to be responsible for the different product profiles of high- and low-artemisinin production chemotypes. Through analysis of their tissue-specific expression and comparison of their sequences with those of other GAOs, it was discovered that one copy of AMO (AMOHAP) exhibits a different transcript compared to the reported artemisinin biosynthetic genes and shows more sequence similarity to other GAOs in the catalytic regions. Furthermore, in a subsequent in vitro enzymatic assay, the recombinant protein of AMOHAP unequivocally demonstrated GAO activity. This result clearly indicates that AMOHAP is a GAO rather than an AMO and that its promiscuous activity on amorpha-4,11-diene has led to its misidentification as an AMO in previous studies. In addition, the divergent expression pattern of AMOHAP compared to that of the upstream germacrene A synthase may have contributed to the abolishment of costunolide biosynthesis in A. annua. Our findings reveal a complex evolutionary landscape in which the emergence of a new metabolic pathway replaces an ancestral one.

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“…In addition, there are sesquiterpene synthases that do not undergo cyclization and produce acyclic sesquiterpenes, such as (E)-β-farnesene synthase from Artemisia annua [ 19 ]. A review has been published summarizing the discovery and catalytic process of ADS, it also described the application of ADS in metabolic engineering and synthetic biology and what important role sesquiterpene synthases played in the evolutionary development of artemisinin [ 20 ]. Including the following information, there was 36% protein similarity between ADS and tobacco 5-epi-aristolochene synthase (TEAS) as well as showed 41% amino acid sequence similarity with cotton ( +)-δ-cadinene synthase.…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis reveals the inconsistency of amorpha-4,11-diene synthase, a key enzyme in the artemisinin synthesis pathway, in asteraceae

et al. 2023

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Background Amorpha-4,11-diene synthase (ADS) is a key enzyme in the artemisinin biosynthetic pathway. ADS promotes the first step of artemisinin synthesis by cyclizing faresyl pyrophosphate to synthesize the sesquiterpene product amorpha-4,11-diene. Thanks to the continuous improvement of genomic information, its evolutionary trace can be analyzed in a genome view. Methods Phylogenetic analysis was used to identify ADS-like genes in other Asteraceae. Gene structure and motif analysis was used to analyze the structural similarity of these identified genes. Heterologous expression and GC–MS analysis were performed to determine whether the functions of ADS and Cna4666 are consistent. Validation of ADS genes evolutionary trajectories was achieved by selective pressure and synteny analysis. Result In this study, we extracted 8 ADS genes from the Artemisia annua L. genome annotation and 121 ADS similar genes from the genomes of Artemisia annua L. and other plants in the Asteraceae, and further exploring their evolutionary relationship. Phylogenetic analysis showed that the genes most closely related to ADS genes were found in the genome of Chrysanthemum nankingense. Among them, the gene structure and motif composition of Cna4666 is very similar to ADS, we wondered whether it has the potential to synthesize amorpha-4,11-diene. Therefore, we extracted the products of recombinant p0_ADS.1 and Cna4666 proteins by HS-SPME combined with GC–MS analysis, the results indicate that Cna4666 is an α-bisabolol synthase, which cannot synthesize amorpha-4,11-diene. Through synteny analysis, we did not find collinear blocks of ADS genes in the Helianthus annuus and C. nankingense genomes. Furthermore, Ka/Ks ratios indicated that the evolution of ADS genes from their similar genes principally underwent purifying selection, and there was a strong positive selection between ADS genes. Conclusions This study proved that ADS is a multi-copy gene in Artemisia annua L., and they are not widely distributed in Asteraceae. The data will increase our understanding of the evolutionary selection pressure on ADS genes. The results suggest that ADS genes are subject to strong positive selection internally, and it is possible that they are a recently evolved gene in the Artemisia.

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Amorpha-4,11-diene synthase: a key enzyme in artemisinin biosynthesis and engineering

Cited by 5 publications

References 97 publications

Functions of Representative Terpenoids and Their Biosynthesis Mechanisms in Medicinal Plants

Functions of Representative Terpenoids and Their Biosynthesis Mechanisms in Medicinal Plants

Functional analysis of CYP71AV1 reveals the evolutionary landscape of artemisinin biosynthesis

Genetic analysis reveals the inconsistency of amorpha-4,11-diene synthase, a key enzyme in the artemisinin synthesis pathway, in asteraceae

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