Abstract
Polygonum minus (syn. Persicaria minor) is a herbal plant that is well known for producing sesquiterpenes, which contribute to its flavour and fragrance. This study describes the cloning and functional characterisation of PmSTPS1 and PmSTPS2, two sesquiterpene synthase genes that were identified from P. minus transcriptome data mining. The full-length sequences of the PmSTPS1 and PmSTPS2 genes were expressed in the E. coli pQE-2 expression vector. The sizes of PmSTPS1 and PmSTPS2 were 1098 bp and 1967 bp, respectively, with open reading frames (ORF) of 1047 and 1695 bp and encoding polypeptides of 348 and 564 amino acids, respectively. The proteins consist of three conserved motifs, namely, Asp-rich substrate binding (DDxxD), metal binding residues (NSE/DTE), and cytoplasmic ER retention (RxR), as well as the terpene synthase family N-terminal domain and C-terminal metal-binding domain. From the in vitro enzyme assays, using the farnesyl pyrophosphate (FPP) substrate, the PmSTPS1 enzyme produced multiple acyclic sesquiterpenes of β-farnesene, α-farnesene, and farnesol, while the PmSTPS2 enzyme produced an additional nerolidol as a final product. The results confirmed the roles of PmSTPS1 and PmSTPS2 in the biosynthesis pathway of P. minus, to produce aromatic sesquiterpenes.
Highlights
Over the last 25 years, nearly 65,000 chemical structures of terpenoids have been discovered, making terpenoids the class of natural products with the greatest structural diversity [1,2]
There are two major pathways involved in the biosynthesis of terpenoids, namely, the mevalonate (MVA) pathway, which is primarily found in eukaryotes, and the methylerythritol phosphate (MEP)
The open reading frames (ORF) of PmSTPS1 started from the nucleotide position at 25 and ended at position 1071
Summary
Over the last 25 years, nearly 65,000 chemical structures of terpenoids have been discovered, making terpenoids the class of natural products with the greatest structural diversity [1,2]. Terpenoids are involved in a variety of important functions in regulating plant growth (especially for terpenoid lactones) and play an ecological role in attracting pollinators [3]. Terpenoids are grouped into different classes based on the number of 5-carbon building blocks [4,5,6,7]. All terpenoids are derived from the common phosphorylated five-carbon (C5) building units, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) [8]. There are two major pathways involved in the biosynthesis of terpenoids, namely, the mevalonate (MVA) pathway, which is primarily found in eukaryotes, and the methylerythritol phosphate (MEP). Pathway (non-mevalonate pathway), which is primarily found in prokaryotes and plant chloroplasts [9,10,11].
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