GGPP

Diterpenoids are derived biosynthetically from geranylgeranyl diphosphate (GGPP), which is itself biosynthesized by reaction of farnesvl diphosphate with isopentenyl diphosphate. Show the structure of GGPP, and propose a mechanism for its biosynthesis horn FPP and IPP. 

IPP and its DMAPP structural isomer are produced from glycolytic products by the methyl erythritol phosphate (MEP) pathway (Figure 5.3.1, Pathway 1). These isoprene units are condensed in a stepwise fashion to form the precursor to all carotenoids, geranylgeranyl di-phosphate (GGPP). GGPP is not solely metabolized to make carotenoids, but is a precursor for many other primary and secondary metab-... 

FIGURE 5.3.1 Parts of the isoprenoid pathways to carotenoids. 1 = MEP pathway. 2 = GGPP synthesis. 3 = Carotenoid biosynthetic pathway. 4 = Carotenoid degradative pathways. Enzyme abbreviations and enzyme activities are defined in Table 5.3.1. 

In plant plastids, GGPP is formed from products of glycolysis and is eight enzymatic steps away from central glucose metabolism. The MEP pathway (reviewed in recent literature - ) operates in plastids in plants and is a preferred source (non-mevalonate) of phosphate-activated prenyl units (IPPs) for plastid iso-prenoid accumulation, such as the phytol tail of chlorophyll, the backbones of carotenoids, and the cores of monoterpenes such as menthol, hnalool, and iridoids, diterpenes such as taxadiene, and the side chains of bioactive prenylated terpenophe-nolics such as humulone, lupulone, and xanthohumol. The mevalonic pathway to IPP that operates in the cytoplasm is the source of the carbon chains in isoprenes such as the polyisoprene, rubber, and the sesquiterpenes such as caryophyllene. 

GGPPS functions as part of a complex metabolon. In the plastid, as shown in Capsicum chromoplasts," GGPPS is a homodimer and associated but not integral to the plastid envelope. GGPPS is also associated with the next enzyme in the pathway as part of a holoenzyme complex." " ... 

The first C40 carotenoid, phytoene, is produced by head-to-head condensation of two GGPPs by an enzyme that shares homology to GGPPS and squalene synthases... 

Careful empirical selection of the expression platform for carotenogenesis has included selection of the best strains for stability and degree of accumulation and the selection of compatible drug-resistance combinations and low copy number polycistronic plasmids to enhance product accumulation by decrease of metabolic burden." 5 Matthews and Wurtzel discussed culture and induction conditions - that have been explored in most studies. Most efforts to engineer carotenoid biosynthesis in E. coli focused on the genes and enzymes of the pathway and had a modest effect on improved accumulation. For example, substitution and over-expression of a GGPPS that uses IPP directly (discussed in... 

Section 5.3.2.3) combined with over-expression of IPPI resulted in enhanced astaxanthin accnmnlation to 1.4 mg/g dry cell weight (DCW). Further increases to 45 mg/g DCW were obtained by random mutagenesis of GGPPS, perhaps by altering enzyme response to substrate-level feedback inhibition. 

Fig. 11.3 Biosynthesis of the homoterpenes (a) DMNT (4,8-dimethyl-l,3,7-nonatriene) and (b) TMTT (4,8,12-trimethyltrideca-l,3,7,l 1-tetraene). (b) Two putative reaction mechanisms for the conversion of (ii,Zi)-geranyUinalool to TMTT catalyzed by a P450 enzyme activity are shown. FPP, farnesyl diphosphate GGPP, geranylgeranyl diphosphate...

Fig. 2. Schematic representation of paclitaxel biosynthesis. Dimethylallyl-diphosphate and isopentenyl-diphosphate are condensed through geranylgeranyl diphosphate synthase activity to render geranylgeranyl-diphosphate (GGPP). GGPP is converted into taxa-4(5), 11 (12)-diene in a reaction catalyzed by the taxane synthase (TS). A series of reactions catalyzed by cytochrome P450 monoxygenases lead to the production of a taxane intermediate that is further converted to baccatin III through enzymes-driven oxidation and oxetane ring formation. The side chain moiety of paclitaxel is derived from L-phenylalanine. Three consecutive arrows mean multiple steps. Ac, acetyl Bz, benzoyl.

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