Living plants are solar-powered biochemical and biosynthetic laboratory which

manufactures both primary and secondary metabolites from air, water, minerals
and sunlight.
The primary metabolites like sugars, amino acids and fatty acids that are needed
for general growth and physiological development of plant are widely distributed
in nature and are also utilized as food by man.
The secondary metabolites such as alkaloids, glycosides, flavonoids, volatile oils
etc are biosynthetically derived from primary metabolites. They represent
chemical adaptations to environmental stresses, or serve as defensive, protective or
offensive chemicals against microorganisms, insects and higher herbivorous
predators. They are some times considered as waste or secretory products of plant
metabolism and are of pharmaceutical importance.
A primary metabolite is directly involved in the normal growth, development, and
reproduction.
A secondary metabolite is not directly involved in those processes, but usually has
important ecological function.

MEVALONIC ACID PATHWAY

Biosynthesis of Mevalonic Acid

A molecule of acetyl-CoA undergoes a Claisen condensation with a molecule of
malonyl-CoA (as in fatty acid biosynthesis; see Biochemistry of Yeast
Fermentation – Fatty Acids) to form acetoacetyl-CoA which reacts with a third
molecule of manonyl-CoA in an Aldol Condensation to give 3-hydroxy-3-
methylglutaryl-CoA (HMG-CoA). Reduction with HMG-CoA reductase yields
mevaldic acid which is reduced again to mevalonic acid.
Conversion of Mevalonic acid to Isopentenyl pyrophosphate (3-methylbut-3-
enylpyrophosphate)
Mevalonate is converted into 3-phospho-5-pyrophosphomevalonate by three
consecutive phosphorylations. This labile intermediate loses CO2 and phosphate
and yields 3-isopentenyl pyrophosphate (IPP).

Isomerization Of Isopentenyl Pyrophosphate And Prenyl Transferase

Reactions
IPP is isomerized to dimethylallyl pyrophosphate (DMAPP) with IPP isomerase.
The isomerization involves addition of a proton to the sp2 carbon of isopentenyl
pyrophosphate and generation of the more stable tertiary carbocation. Elimination
of a proton yields the most stable alkene (trisubstituted, Zaitev’s Rule).

Next IPP reacts with DMAPP to give geranyl pyrophosphate (GPP). In the first
step of the reaction, IPP acts as a nucleophile and displaces a pyrophosphate group
from DMAPP. Pyrophosphate is an excellent leaving group. Its four OH groups
have pKa values of 0.9, 2.0, 6.6, and 9.4. Therefore three of the four groups will
be primarily in their basic forms at physiological pH (pH 7.3). A proton is
removed in the next step, again yielding the most stable alkene. GPP rearranges to
give the monoterpenes. GPP reacts with a second molecule of IPP to produce
farnesyl pyrophosphate (FPP). FPP is the precursor to the sesquiterpenoids, and
the steroids. FPP reacts with a third molecule of IPP to produce geranylgeranyl
pyrophosphate (GGPP). GGPP is the precursor of the carotenoids.
Significance of Mevalonic acid pathway:
Isoprenoid Compounds:
The isoprenoids are built up of C5 isoprene units and the nomenclature of the main
classes reflects the number of isoprene units present (Table 1).
Monoterpenes:
Acyclic Monoterpenes:
In monoterpene diphosphate, geranyl PP, the double bond is trans (E). Linalyl PP
and neryl PP are isomers of geranyl PP, and are likely to be formed from geranyl
PP by ionization to the allylic cation, which can thus allow a change in attachment
of the diphosphate group (to the tertiary carbon in linalyl PP) or a change in
stereochemistry at the double bond (to Z in neryl PP) (Figure 5.9). These three
compounds can give rise to a range of linear monoterpenes found as components
of volatile oils used in flavouring and perfumery (Figure 5.10). E.g.: Nerol,
Geraniol, citronellol, linalool, hotrienol
MonoCyclic Monoterpenes:
Cyclic monoterpenes can not be expected to occur with the precursor geranyl
diphosphate, because the E stereochemistry of the double bond being unfavourable
for ring formation (Figure 5.9). Neryl PP or linalyl PP, however, do have
favourable stereochemistry, and either or both of these would seem more
immediate precursors of the monocyclic menthane system, formation of which
could be represented as shown in Figure 5.12, generating a carbocation (termed
menthyl or α-terpinyl) having the menthane skeleton. It has been found that
monoterpene cyclase enzymes are able to accept all three diphosphates, with
linalyl PP being the best substrate, and it appears they have the ability to isomerize
the substrates initially as well as to cyclize them. It is convenient therefore to
consider the species involved in the cyclization as the delocalized allylic cation
tightly bound to the diphosphate anion, and bond formation follows due to the
proximity of the π-electrons of the double bond (Figure 5.12).
The menthyl cation could be quenched by attack of water, in which case the
alcohol α-terpineol would be formed, or it could lose a proton to give limonene
(Figure 5.13). The menthyl cation, although it is a tertiary, may be converted by a
1,3-hydride shift into a favourable resonance stabilized allylic cation (Figure
5.13). This allows the formation of α- and β-phellandrenes by loss of a proton
from the phellandryl carbocation. E.g. of monocyclic terpenoids are p-cymene,
thymol, carvacrol, menthol, isomenthol, neomenthol, menthone, isomenthone,
neoisomenthone, pulegone, isopulegone, carveol, carvone, piperitone,
piperitenone, isopiperitenone, isopiperitenol,
Dicyclic Monoterpenes:

Alternatively, folding the cationic side-chain towards the double bond (via the
surface characteristics of the enzyme) would allow a repeat of the cyclization
mechanism, and produce bicyclic bornyl and pinyl cations, according to which end
of the double bond was involved in forming the new bonds (Figure 5.14). Borneol
would result from quenching of the bornyl cation with water, and then oxidation of
the secondary alcohol could generate the ketone camphor. As an alternative to
discharging the positive charge by adding a nucleophile, loss of a proton would
generate an alkene. Thus α-pinene and β-pinene arise by loss of different protons
from the pinyl cation, producing the double bonds as cyclic or exocyclic
respectively. A less common termination step involving loss of a proton is the
formation of a cyclopropane ring as exemplified by 3-carene and generation of the
carane skeleton.
The bicyclic pinyl cation, with a strained four membered ring, rearranges to the
less strained five-membered fenchyl cation (Figure 5.14), a change which
presumably more than makes up for the unfavourable tertiary to secondary
carbocation transformation. This produces the fenchane skeleton, exemplified by
fenchol and fenchone. The isocamphyl tertiary carbocation is formed from the
bornyl secondary carbocation by a Wagner–Meerwein rearrangement, and so leads
to camphene. A hydride shift converting the menthyl cation into the terpinen-4-yl
cation only changes one tertiary carbocation system for another, but allows
formation of α-terpinene, γ-terpinene, and the α-terpineol isomer, terpinen-4-ol.
A further cyclization reaction on the terpinen-4-yl cation generates the thujane
skeleton, e.g. sabinene and thujone.

Sesquiterpenes:
FPP can then give rise to linear and cyclic sesquiterpenes. Because of the
increased chain length and additional double bond, the number of possible
cyclization modes is also increased, and a huge range of mono-, bi-, and tri-cyclic
structures can result. The stereochemistry of the double bond nearest the
diphosphate can adopt an E configuration (as in FPP), or a Z configuration via
ionization, as found with geranyl/neryl PP (Figure 5.26). In some systems, the
tertiary diphosphate nerolidyl PP (compare linalyl PP, page 172) has been
implicated as a more immediate precursor than farnesyl PP (Figure 5.26). This
allows different possibilities for folding the carbon chain, dictated of course by the
enzyme involved, and cyclization by electrophilic attack on to an appropriate
double bond.

E,E-Farnasyl cation, E,Z-Farnasyl cation and nerolidyl cation give

rise to matricin, α -santonin, humulene, thapsigargin, bisabolene,
α -bisabolol, caryophyllene, artemisinic acid, α -cardinene,
zingiberene, sesqui-phellandrene, thapsigargin, parthenolide,
chamazulene.
Diterpenoids:
The diterpenes arise from geranylgeranyl diphosphate (GGPP). One of the
simplest and most important of the diterpenes is phytol, a reduced form of
geranylgeraniol, which forms the lipophilic side-chain of the chlorophylls, e.g.
chlorophyll a. E.g. of diterpenoids are

Triterpenoids:
Two molecules of farnesyl PP are joined tail to tail to yield the
hydrocarbon squalene which act as precursor of triterpenes and
steroids.
Cyclization of squalene is via the intermediate squalene-2,3-
oxide (Figure 5.55), produced in a reaction catalysed by a
flavoprotein requiring O2 and NADPH cofactors.
If squalene oxide is suitably positioned and folded on the enzyme
surface, the polycyclic triterpene structures formed can be
rationalized in terms of a series of cyclizations, followed by a
sequence of concerted Wagner–Meerwein migrations of methyls
and hydrides (Figure 5.55).
The cyclizations are carbocation mediated and proceed in a step-
wise sequence (Figure 5.56).
Examples of triterpenoids prepared from squalene are euphol, lupeol,
dammarenediol, taraxasterol, α -amyrin, β -amyrin. E.g. of triterpenoid saponin
are hopan-22-ol, tetrahymanol, quillaic acid, barringtogenol, glycyrrhizic acid,
liquiritin, isoliquiritin, carbenoxolone sodium, panaxadiol, panaxatriol, oleanolic
acid, syringin.

Tetraterpenes:
Formation of the tetraterpene skeleton, e.g. phytoene, involves
tail-to-tail coupling of two molecules of geranylgeranyl
diphosphate (GGPP) in a sequence
essentially analogous to that seen for squalene and triterpenes
(Figure 5.67).
For phytoene biosynthesis, a proton is lost, generating a double
bond in the centre of the molecule, and thus a short conjugated
chain is developed. This triene system prevents the type of
cyclization seen with squalene. Conjugation is extended then by a
sequence of desaturation reactions, removing pairs of hydrogens
alternately from each side of the triene system, giving eventually
lycopene (Figure 5.67). The tetraterpenes are represented by
only one group of compounds, the carotenoids. E.g.
Lycopene, α -carotene, β -carotene, γ -carotene,
zeathanthin, lutein, violaxanthin, capsanthin, fucoxanthin,
astaxanthin.
Steroids:
The steroids are modified triterpenoids containing the tetracyclic ring system of
lanosterol but lacking the three methyl groups at C-4 and C-14.