Deletion and tandem duplications of biosynthetic genes drive the diversity of triterpenoids in Aralia elata
Deletion and tandem duplications of biosynthetic genes drive the diversity of triterpenoids in Aralia elata
Tandem duplication of genes drives the diversity of oleanane-type pentacyclic triterpenes in A. elata The colinearity relationship of CYP72A (a), CSLM (b), and two UGT73 (c and d) tandem duplication regions between A. elata and P. notoginseng. The syntenic blocks are connected by gray lines. The heatmap shows the FPKM values (normalized) of the colinear gene in different tissues of A. elata. The green circles represent the tandem-repeat enzyme-modification positions. The blue bar represents the genes on the forward strand of the genome; the green bar represents genes on the reverse strand. Source data are provided as a Source Data file.
De novo biosynthesis of oleanane-type pentacyclic triterpenes in yeast a The strain WEA expressed AeBAS, AeCYP716A354, and AeCYP716A355 and overexpressed the MVA and early sterol-pathway genes tHMG1, ERG20, ERG9, and ERG1. Diagram showing the echinocystic acid-type triterpenoid biosynthetic pathway. b LC–MS analysis of metabolites from strains engineered for pentacyclic triterpene saponin production derived from β-amyrin and oleanolic acid. Extracted ion chromatograms using m/z values (±0.5) and different numbers to represent saponins with various aralosides, oleanolic acid (m/z 455, 2), echinocystic acid (m/z 471, 3), hederagenin (m/z 471, 4), 16-OH-hederagenin (m/z 487, 5), oleanolic acid 28-O-glucopyranosyl ester (m/z 617, 6), calenduloside E (m/z 631, 7), zingibroside R1 (m/z 793, 8), chikusetsusaponin IVa (m/z 793, 9), echinocystic acid 28-O-glucopyranosyl ester (m/z 633, 11), 3-O-glucuronopyranoside echinocystic acid (m/z 647, 12), echinocystic acid 28-O-glucopyranosyl-3-O-glucuronopyranoside (m/z 809, 14), and echinocystic acid 3-O-glucuronopyranoside-6 → 1-glucoside (m/z 809, 13).
Overview of araloside structures and their biosynthesis in A. elata The light-yellow box indicates the cyclization of 2,3-oxidosqualene to β-amyrin and CYP-catalyzed hydroxylation of triterpenoids. The boxes in other colors represent the glycosylation of different triterpene backbones. Brown letters indicate hydroxylation reactions catalyzed by cytochrome P450 monooxygenases (P450s); blue letters indicate glycosylation reactions catalyzed by cellulose synthase-like glycosyltransferase (CSL); green letters indicate C28 glycosylation reactions catalyzed by UDP-dependent glycosyltransferases (UGTs); and purple letters indicate C3–2 glycosylation reactions catalyzed by UGTs. Reaction sites are indicated in the corresponding colors. Broken arrows indicate putative araloside biosynthesis steps.
The species and metabolic evolution model of Araliaceae a Schematic diagram of divergence times in Apiales. The black points indicate the divergence-time nodes. b The pink and blue boxes represent the metabolites of Panax and A. elata, respectively. Genes involved in the alaroside biosynthetic pathway. The black arrow shows the cyclization of 2,3-oxidosqualene to β-amyrin catalyzed by BAS, the purple arrow shows the hydroxylation at the corresponding site catalyzed by P450, the blue arrow shows the glucuronosylation at the corresponding site by CSL, and the green arrows show the glycosylation reactions by the corresponding UGTs. The blank arrows indicate the pseudogenes in the tandem repeat that are not involved in the alaroside biosynthetic pathway. The compounds in dashed boxes represent the specific triterpenoids in Panax and A. elata. TTs tetracyclic triterpenoids, PTs pentacyclic triterpenoids.
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