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e secondary metabolite synthesis pathways. Putative genes related to the biosynthesis of glycyrrhizin In this study, our primary goal was to identify genes involved in the glycyrrhizin biosynthetic pathway, The biosynthesis of gly cyrrhizin involves the synthesis of dimethylallyl diphosphate and isopentenyl TCID diphosphate, the biochemically active isoprene units of all terpe noids, This step is followed by the synthesis of the triterpene skeleton, also known as B amyrin, and then by a series of oxidative reactions and glucuronyla tions, which produce glycyrrhizin. The precise order of the intermediate products is still unknown, In the early stage of active isoprene unit formation, plants have the ability to produce DMAPP and IPP using two pathways, the mevalonate pathway and the methylerythritol phosphate pathway, In plants, these two pathways appear to be separate.
enzymes of the MVA pathway are found in the cytosol, whereas enzymes of the MEP pathway are localized in plastids. Triterpenoids are known to be formed by the MVA pathway because they are cytosolic products. How ever, there are examples where the two pathways can act TCID cooperatively to create a molecule, No progress has been made toward determining the precise source of iso prene units in glycyrrhizin biosynthesis. Using a BLAST search against the SwissProt and KEGG databases, we found the genes encoding all of the enzymes from both of these two pathways in the EST database, except for mevalonate kinase, which is located in the MVA pathway, and DXP synthase, which is located in the MEP pathway.
In this study, we found all of the putative genes encoding IU1 the enzymes involved in the triterpene skeleton B amyrin synthesis step. farnesyl diphosphate synthase, squalene synthase, squalene monooxygenase and B amyrin synthase, The enzymes involved in the biosyntheses of the isoprene unit and the triterpene skel eton are listed in Table 3. A list of putative unigenes involved in the glycyrrhizin biosynthetic pathway is shown in Additional file 3. Cytochrome P450 and glycosyltransferase Glycyrrhizin is derived from the triterpene B amyrin, which is an initial from product of thethe cyclization of 2, 3 oxidosqualene. The subsequent steps in glycyrrhizin biosynthesis include a series of oxidative and glycosyl transfer reactions.
Plant morphology We have little knowledge of the later GDC-0152 steps in the glycyrrhizin biosynthetic TCID pathway, which include multiple oxidation and glycosylation steps that are catalyzed by enzymes from the cytochrome P450 and glycosyltransferase superfamilies, respectively. Cytochrome P450 is a very large and diverse superfam ily of hemoproteins that are found in all higher organisms, Plant P450s catalyze many different reactions involved in the biosynthesis of secondary metabolites, including terpenoids, Some members of the CYP88 and CYP93 families have been shown to act on B amyrin or related triterpene substrates with unique reaction specificities, Thus far, all known cytochrome P450s that act on triterpenes and sterols have been classi fied into two clans. the CYP71 clan and the CYP85 clan, which includes CYP93 and CYP88, respectively, GDC-0152 Only two CYP genes of G.
uralensis have been identified, An organ TCID specific transcript profil ing approach was used in other studies to identify CYP88D6, which catalyzes the oxidation of B amyrin at C 11 to produce 11 oxo B amyrin in the glycyrrhizin bio synthetic pathway. The expression profile of CYP88D6 was consistent with the organ specific accumulation pat tern of glycyrrhizin, a higher level of expression was seen in the root than in the stem and leaf. By mining the EST database, we found 125 unigenes anno tated GDC-0152 as putative CYP genes, which were further classified into 32 CYP families and 47 subfamilies, To narrow down the candidate cytochrome P450s, these unigenes were further screened according to their classification. In the candidate P450 dataset, two unige nes were annotated as CYP88, while six unige nes were annotated as CYP93. In tota