is isomerized to the other isoprene unit, DMAPP, by an isomerase enzyme which incorporates a proton

from water onto C-4 and stereospecifically removes the pro-R proton (HR) from C-2. This reaction is
used to provide the two compounds in the amounts required for further metabolism. Two different
types of isomerase enzyme have been distinguished: a type I enzyme requiring a divalent metal ion and
a type II enzyme that requires a divalent metal ion together with FMN for activity. Both enzymes appear
to employ a protonation–deprotonation mechanism. The conversion of IPP into DMAPP generates a
reactive electrophile and, therefore, a good alkylating agent. DMAPP possesses a good leaving group,
the diphosphate, and can ionize readily to yield 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 OPP resonance-stabilized allylic cation DMAPP carbocation formation
Note: when using this representation of the allylic cation, do not forget it contains a double bond Figure
5.5 the electrophilic DMAPP. These differing reactivities are the basis of terpenoid biosynthesis, and
carbocations feature strongly in mechanistic rationalizations of the pathways

lyceraldehyde 3-phosphate are used in the production of MEP; the pyruvate carboxyl is lost in this
process (Figure 5.6). Thiamine diphosphate-mediated decarboxylation of pyruvate (compare page 23)
produces an acetaldehyde-equivalent bound in the form of an enamine. This reacts as a nucleophile in
an addition reaction with the glyceraldehyde 3-phosphate. Subsequent release from the TPP carrier
generates 1-deoxy-d-xylulose 5-phosphate (deoxyxylulose phosphate), which is transformed into MEP
by a rearrangement process. This has been shown to involve a reverse aldol–aldol sequence (Figure 5.6),
coupled with a reduction. A single enzyme catalyses these skeletal rearrangement and reduction
reactions without release of any intermediate; the product now contains the branched-chain system
equivalent to the isoprene unit. Reaction of MEP with cytidine triphosphate (CTP) produces a cytidine
diphospho derivative (compare uridine diphosphoglucose in glucosylation, page 31), which is then
phosphorylated via ATP. The resultant 2-phosphate is then converted into a cyclic phosphoanhydride
with loss of cytidine phosphate. The subsequent steps leading to IPP and DMAPP are the least
understood part of the pathway. Gene methodology has shown that two enzymes are involved, the first
producing 4-hydroxy-3-methylbut-2-enyl diphosphate and the second converting this into
predominantly IPP, but also DMAPP. Both steps are reductive in nature, but mechanisms are yet to be
elucidated. The formation of both IPP and DMAPP (ratios are typically in the region 5:1 to 4:1) is
suggested to involve a delocalized allylic system (radical or anion, shown in Figure 5.6 as an anion), with
protons being supplied by water. Although this pathway coproduces IPP and DMAPP, isomerism of IPP
to DMAPP as in the mevalonate pathway is also possible to balance the pool sizes of these
intermediates. Whether the mevalonate pathway or the MEP pathway supplies isoprene units for the
biosynthesis of a particular terpenoid must be established experimentally. This can be determined from
the results of feeding [1-13C]-d-glucose as precursor; this leads to different labelling patterns in the
isoprene unit according to the pathway operating (Figure 5.7). Animals and fungi appear to lack the MEP
pathway, so utilize the mevalonate pathway exclusively. The MEP pathway is present in plants, algae,
and most bacteria. Plants and some bacteria are equipped with and employ both pathways, often
concurrently. In plants, the two pathways appear to be compartmentalized, so that the mevalonate
pathway enzymes are localized in the cytosol, whereas the MEP pathway enzymes are found in
chloroplasts. The cytosolic pool of IPP serves as a precursor of C15 derivatives (farnesyl PP (FPP) and
sesquiterpenes; see Figure 5.2), and in due course triterpenoids and steroids (2 × FPP). Accordingly,
triterpenoids and steroids, and some sesquiterpenoids (cytosolic products) are formed by the
mevalonate pathway, whilst most other terpenoids (C10, C20, C40) are formed in the chloroplasts and
are MEP derived. Of course there are exceptions. There are also examples where the two pathways can
supply different portions of a molecule, or where there is exchange of late-stage common intermediates
between the two pathways (cross-talk), resulting in a contribution of isoprene units from each pathway.
In the following part of this chapter, these complications will not be considered further, and in most
cases there is no need to consider the precise source of the isoprene units. An area of special
pharmacological interest where the early pathway is of particular concern is steroid biosynthesis, which
appears to be from mevalonate in the vast majority of organisms. Thus, inhibitors of mevalonate
pathway enzymes will reduce steroid production in plants, but will not affect the formation of
terpenoids derived via MEP. Equally, it is possible to inhibit terpenoid production without affecting
steroid formation by the use of MEP pathway inhibitors, such as the antibiotic fosmidomycin from
Streptomyces lavendulae. This acts as an analogue of the rearrangement intermediate in the reaction
catalysed by MEP synthase (Figure 5.6). Enzymes of the MEP pathway are attractive targets for
development of drugs against microbial diseases such as malaria or tuberculosis, since the MEP pathway
is utilized by the pathogen but is not present in humans. Regulation of cholesterol production in humans
is an important health concern (see page 251); the widely used statin drugs are specific inhibitors of the
mevalonate pathway enzyme HMG-CoA reductase (see page 98)