WIWILEY B Viteelturery Natural oy > Products _ A Biosynthetic Approach eV eae170 reduction of eldetone evento aid peemeia | Siphexphate - NS ‘orr RT a ae i eee aaa n® ®) ° Aven = we SOL + ma scetoacery COA wo Sing — we SEP mevaliie id ‘THE MEVALONATE AND DEOXYXYLULOSE PHOSPHATE PATHWAYS stereoupeeific allel reaction ‘are omapen tinge ‘enayime-boune wntc-con | apen ‘fain of its (iter to aldehyde mevaide wid stereospecific llstic favre re mies imexny ay —— Figure 5.4 phosphorylation of the primary alcohol group. Two different ATP-dependent enzymes are involved, resulting in mevalonic acid diphosphate, and decarboxylation/dehydration then follow to give IPP. Whilst a third molecule of ATP is required for this last transformation, there is no evidence for phosphorylation of the tertiary hydroxyl, though this would convert the hydroxyl into a better leaving group. Perhaps ATP assists the loss of the hydroxyl as shown in Figure 5.4. IPP is isomerized to the other isoprene unit, DMAPP, by an isomerase enzyme which stercospecifically removes the pro-R proton (Hy) from C-2, and incorporates a proton from water on to C-4, Whilst the isomerization is reversible, the equilibrium lies heavily on the side of DMAPP. This conversion generates a reactive electrophile and therefore a ‘good alkylating agent. DMAP possesses a good Ieaving group, the diphosphate, and can yield via an Syl process an allylic carbocation which is stabilized by charge delocalization (Figure 5.5). In contrast, IPP with its terminal double bond is more likely to act as a nucleophile, especially towards the electrophilic DMAPP. These differing reactivities are the basis of terpenoid biosynthesis, and carbocations feature strongly in mechanistic rationalizations of the pathway’. L-Deoxy-D-xylulose 5-phosphate is formed from the glycolytic pathway intermediates pyruvic acid and glyceraldehyde 3-phosphate with the loss of the pyruvate carboxyl (Figure 5.6). Thiamine diphosphatie-mediated decarboxylation of pyruvate Ag, Ser Mare [Street None: whem using this Trepresenuann ofthe forget he dole ond AE -- Je, ~- 12 Figure 5.5‘THE MEVALONATE AND DEOXYXYLULOSE PHOSPHATE PATHWAYS crinel Be Figure 5.3 the steps leading to and from mevalonic acid were gradually detailed in a series of painstakingly exe- cuted experiments. For many years, the early parts of the mevalonate pathway were believed to be common to the whole range of natural terpenoid derivatives, but it has since been discovered that an alternative pathway to IPP and DMAPP exists, via deoxyxylulose phosphate. and that this pathway is probably more widely utilized in nature than is the mevalonate pathway. This pathway is also referred {0 as the mevalonate-independent pathway or the methylerythritol phosphate pathway Three molecules of acetyl-coenzyme A ate used to form mevalonic acid. Two molecules combine initially in a Claisen condensation to give acetoacetyl-CoA, and a third is porated via a stereospecific aldol addition giving the branched-chain ester B-hydroxy-f- methylglutaryl-CoA (HMG-CoA) (Figure 5.4). This third acetyl-CoA molecule appears to be bound to the enzyme via a thiol group, and thi linkage is subsequently hydrolysed to form the free acid group of HMG-CoA. In the acetate pathway, an acetoacetic acid thioester (bound ( the acyl carrier protein) would have been formed using the more nucleophilic thioester of malonic acid. The mevalonate pathway does not use mal- onyl derivatives and it thus diverges from the acetate pathway at the very first step. In the second step, it should be noted that, on purely chemical grounds. acetoacetyCoA is the more acidic substrate, and might be expected to act as the nucleophile rather than the third acetyl-CoA molecule. The enzyme thus achieves what is a less favourable reaction. The conversion of HMG- CoA imo (32)-MVA involves a two-step reduc tion of the thioester group to a primary alcohol, and provides an essentially irreversible and rate- limiting transformation. Drug-mediated inhibition of this enzyme (HMG-CoA reductase) can be used to regulate the biosy we and ultimately of the steroid cholesterol (see statins, page 112). The six-carbon compound MVA is transformed imo the five-carbon phosphorylated isoprene units in a series of reactions, beginning with thesis of mevalonwee ce capper ructures when rearrangement reac- place, e.g. steroids, where in addi- bons have been lost. Neverthele: 3 are formed via regular terpenoid iatural products contain terpenoid r molecules, in combination with 3 derived from other sources, such and shikimate pathways. Many olics, and vitamins discussed in re examples of this. A particularly oid fragment in such cases is a Mevalonic acid ~ Aheertaie dimethylallyl PP A \ cer isopentenyl PP to indicate the dimethylallyl su macromolecules like proteins can attaching terpenoid chains. Cystei alkylated with farnesyl or geranyl thereby increasing the lipophilicity and its ability to associate with m« The biochemical isoprene units by two pathways, by way of intern onic acid (MVA) (Figure 5.4) xylulose 5-phosphate (deoxyxylul DXP) (Figure 5.6). Mevalonic aci uct of acetate metabolism, had bee a precursor of the animal sterol « Deoxyxylulose ot phosphate Hemiterpenes (C9) (DMAPP) (Cs) Y APP) (Cs) Cio Monoterpenes (Cio) Iridoids Pp \ Cis, _ Sesquiterpenes (Cis) Iee x2 Ca =| ————+_ Diiterpenes (Cap) IPP \ C2 = + _ Sesterterpenes (C25) Cao Cao Triterpenoids (C10) | Steroids (C1s—Cro) Tetraterpenes (Cao) Carotenoids Figure 5.2