Cloning and Expression Analysis of a Farnesyl Diphosphate Synthase (FPPS) Gene from Chamaemelum nobile

Abstract

Farnesyl diphosphate synthase (FPPS), an isopentenyl transferase, catalyzes the condensation reaction of five carbon isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) to form fifteen carbon farnesyl pyrophosphate (FPP), which is the key precursor for sesquiterpene biosynthesis. In this study, a FPPS gene (CnFPPS) was cloned from Chamaemelum nobile. The full-length cDNA of CnFPPS is 1239 bp and contains an open reading frame (ORF) of 1029 bp encoding 342 amino acids. The theoretical molecular weight and pI of the CnFPPS protein are 39.38 kDa and 5.59, respectively. Multiple alignment analysis showed the protein sequence of CnFPPS had a high homology with FPPS proteins from other plants. The deduced amino acid of CnFPPS contained five conservative domains such as substrate binding pocket, substrate-Mg 2+ binding site, catalytic site, aspartate-rich region 1 and 2, suggesting CnFPPS is one member of FPPS family in C. nobile. Phylogenetic analysis based on the amino acid sequences of FPPSs showed that CnFPPS was closely related to the FPPS of Matricaria chamomilla. The result of qRT-PCR revealed that CnFPPS gene was constitutively expressed in different tissues of C. nobile, with the highest expression in the root. These findings improve the understanding of the synthesis and regulation of the terpenoid compounds at the molecular level and lay a foundation for studying the regulatory functions of CnFPPS in terpenoid biosynthetic pathway in C. nobile.

Agarwood Induction: Current Developments and Future Perspectives

“…These C5 isoprene units will later be channeled into the generation of C15 farnesyl pyrophosphate (FPP) by sequential condensation reactions in the presence of FPP synthase (FPS) (Rohmer, 1999; Yang et al, 2013; Ye et al, 2016). The FPS is one of the key-limiting enzymes responsible for the sesquiterpene biosynthesis (Gaffe et al, 2000; Yang et al, 2013; Liu X. M.et al, 2017). The genes encode for FPS have been cloned from Aquilaria microcarpa ( Am-FaPS-1 ) (Kenmotsu et al, 2011) and Aquilaria sinensis ( AsFPS1 ) (Yang et al, 2013).…”

Agarwood—The Fragrant Molecules of a Wounded Tree

“…These C5 homoallylic isoprenoid precursors on sequential condensation in the presence of C15 farnesyl pyrophosphate synthase generates C15 farnesyl pyrophosphate [ 66 , 69 ]. Researchers such as Yang et al [ 69 ] and Liu et al [ 70 ] have stated that C15 farnesyl pyrophosphate synthase acts as the key limiting factor in the synthesis of sesquiterpenes that are encoded by the gene Am-FaPS- 1 and AsFPS 1 in Aquilaria species. Another set of enzymes, namely sesquiterpene synthase and cytochrome P450 dependent mono-oxygenases, play a pervasive role in the final stage of the sesquiterpene synthesis by oxidative functionalization of the C15 farnesyl pyrophosphate with the cytochrome P450 dependent mono-oxygenases, thus forming a multicyclic scaffold complex.…”

“…Terpenoids are abundantly present in nature, especially in plants, and are known to contain oxygen derivatives that bear a hydroxyl group at the C-3 position [ 77 ]. To date, about three different terpenoids have been identified in agarwood and Aquilaria plant leaves, namely 3-oxo-22-hydroxyhopane, 3b-olean-12-ene-3,23-diol, and hederagenin [ 13 , 70 , 97 ]. Among all the terpenoids known, sesquiterpenoids are the dominant fragrant compounds observed to be present in agarwood and agarwood products with the presence of three isoprene structural units ( Table 3 ) [ 82 , 165 ].…”