Lecture n.

THE SHIKIMIC ACID PATHWAY

 an activated form of gallic acid and has similar reac-
a component of many tannins, which are plant materi- tivity to a CoA thioester. Tannins also contribute to OH
THE SHIKIMIC ACID PATHWAY
als that have been used for thousands of years in the the astringency of foods and beverages, especially tea, OH
tanning of animal hides to make leather because of coffee, and wines, through their strong interaction with
their ability to cross-link protein molecules. Gallotan- salivary proteins. In addition, they have beneficial antiox- O
OH
nins are esters of gallic acid with a polyalcohol, typically idant properties (see also condensed tannins, page 171). OH OH
OH OH O
OH β-glucogallin OH
OH UDPGlc O O
HO HO
OP HO O HO O
E1 OH E2 OH
O HO2C OH OH OH
HO glucose
formally an elimination of phosphoric acid; it actually O O
HO CO2H involves oxidation of the hydroxyl adjacent to the proton 1,6-digalloylglucose
gallic acid β-glucogallin
lost and therefore requires NAD+ cofactor; the carbonyl
PEP DAHP OH is subsequently reduced back to an alcohol
CO2H (hemiketal form)
CO2H E3−5 β-glucogallin (3 ×)
aldol-type CO2H
P O
reaction NAD+ H O
PO PO O O one electron oxidations
H OH OH of phenol groups give OH
E1 E2 resonance-stabilized OH
H HO O OH
HO O H OH HO OH O radicals
HO HO OH
OH OH OH O
aldol-type OH O2
O OH
D-erythrose 4-P DAHP reaction OH
O E6 O
O O OH
O O O O O
CO2H CO2H HO CO2H HO CO2H O O O O
NADPH – H2O NADH O O OH
radical coupling O
HO OH E4 O OH E3 O OH E5 HO OH
HO OH HO OH
OH OH OH OH
OH HO OH
shikimic acid 3-dehydroshikimic 3-dehydroquinic quinic acid HO
acid acid HO HO OH
pentagalloylglucose
dehydration E6 – H2O O oxidation O
and enolization and enolization HO OH
O
OH hydrolysis
CO2H CO2H glucose
E1: DAHP synthase (aroF, aroG, aroH) O O
O
E2: 3-dehydroquinate synthase (aro B) O O O hydrolysis + glucose + gallic acid
E3: 3-dehydroquinase (aro D) O O OH
gallic acid
E4: shikimate dehydrogenase (aroE) O
E5: quinate dehydrogenase HO HO OH +
OH OH
E6: 3-dehydroshikimate dehydratase OH OH
HO OH HO OH O O OH
protocatechuic gallic acid HO2C
acid OH HO OH spontaneous
lactonization
OH
Figure 4.2 CO2H
tellimagrandin II
HO OH HO O O
E1: gallate 1-β-glucosyltransferase OH OH
E2: β-glucogallin O-galloyltransferase
hexahydroxydiphenic acid ellagic acid
E3−5: galloyltransferases
E6: pentagalloylglucose: oxygen oxidoreductase
 transamination:

THE SHIKIMIC ACID PATHWAY

140 Medicinal Natural Products: A Biosynthetic Approach. 3rd Edition CO2H
keto acid amino acid
CO2H

O NH2
nucleophilic attack onto
protonated double bond of PEP H PLP
PEP
CO2H CO2H CO2H
H E6
PO CO2H H H
ATP phenylpyruvic acid L-Phe
decarboxylation, decarboxylation,
aromatization and E2 E3 aromatization and
HO OH E1 PO OH E2 PO O CO2H loss of leaving group loss of leaving group
OH OH OH OP O CO2H O CO2H
CO2H H H
shikimic acid shikimic acid 3-P
– HOP 1,2-elimination of O O
O PLP NH2
phosphoric acid
1,4-elimination of
CO2H CO2H
E7 Aromatic Amino Acids and Phenylpropanoids 145
The Shikimate Pathway:
CO2H phosphoric acid CO2H
O CO2H E1
HN O O FMNH2 OH OH OH
H
Anthranilic acid (Figure 4.5) is an intermediate in the by the multi-enzyme complex tryptophan synthase. Al-
P OH O P OH chorismic acid prephenic acid L-arogenic acid
O CO2H E3 PO O CO2H biosynthetic pathway to the indole-containing aromatic though a precursor of l-tryptophan, anthranilic acid may
OH OH an additional oxidation also be produced by metabolism of tryptophan. Both com-
OH OH amino acid l-tryptophan (Figurestep
4.11). InE4 a sequence
NAD+ E5 NAD+ oxidation means OH is
glyphosate PEP (of alcohol to ketone) means OH retained
pounds feature as building blocks for on decarboxylation
a variety of alkaloid
chorismic acid EPSP of complex reactions, which will not be
is retained on decarboxylation considered in de- and aromatization;
(5-enolpyruvylshikimic acid 3-P) tail, the indole
CO2H structures (see Chapter
CO6).
2H
and ring system isnoformed
aromatization; discrete by incorporating no discrete ketone
E1: shikimate kinase (aro L) E3: chorismate synthase (aro C) ketone from
intermediate is formed diphosphate, with Returning to the main course of the isshikimate
intermediate formed path-
The Shikimate Pathway: Aromatic Amino Acids and Phenylpropanoids 141 two carbon atoms phosphoribosyl O NH
E2: EPSP synthase (aro A) way, a singular rearrangement process occurs, transform-
2
loss of the original
E1: chorismate mutase anthranilate carboxyl. The remaining chorismic acid into prephenic acid (Figure 4.12).
ingPLP
Figure 4.4 elimination of pyruvic acid elimination of pyruvic acid ribosyl carbon dehydratase
E2: prephenate atoms are then removed by a reverse aldol This reaction, a Claisen rearrangement, transfers the
(formally as enolpyruvic acid) (formally as enolpyruvic acid) E3: arogenate dehydratase
reaction, to bedehydrogenase
E4: prephenate replaced on a bound form of indole by PEP-derived
E8 side-chain so that it becomes directly bonded
generates aromatic ring isomerization generates aromatic ring
CO2H those from l-serine,
E5: arogenate which then becomes the side-chain
dehydrogenase to the carbocycle OHand so builds up the basic C6 C3 car-
this is not a simple concertedCOelimination.
2H
OH2
via SN2′ reaction
Despite CO2H
para-hydroxylation CO2H
and the ortho-polyhydroxylation pat- E6:l-tryptophan.
of phenylpyruvate aminotransferase
The
OH
last two steps shown are catalysed bon skeleton of phenylalanine and tyrosine. The reaction
the reaction involving no overall change in oxidation terns contrast withOH the typical meta-hydroxylation OH patterns E7: prephenate aminotransferase 4-hydroxyphenyl- L-Tyr
E8: 4-hydroxyphenylpyruvate pyruvic acid
state, the enzyme requires reduced FMN, which is not characteristic of phenols derived via the acetate pathway aminotransferase
consumed in the reaction, E1 and this coenzyme
O is 2believed
CO H E2 (see page 101),Oand CO in most E3 allow the biosynthetic
cases
2H OH
CO2H CO2H CO2H
to participate
OH via reverse transfer OHof an electron to the origin isochorismic
(acetate or acid
shikimate) of an salicylic
aromatic ring to be Figure 4.13 SN2 reaction
H acid HO
substrate. The biosynthesis of chorismic
4-hydroxybenzoic chorismicacid
acidfrom phos- hydrolysis
deduced by inspection. OP
of NH2 NH CH2OPP H
phoenolpyruvate
acid and d-erythrose 4-phosphate involves enol2,3-Dihydroxybenzoic
ether E8 acid and (in microorgan- CH2OP
E1 E2
OH
L-Gln L-Gln is catalysed in nature by the enzyme chorismate mutase, NADO+ -dependent dehydrogenase N
seven enzyme activities.amination In most using isms, but not in plants;oxidation
prokaryotes, these side-chain see pageof 3-hydroxyl to
161) salicylic acid anthranilic O
enzyme,
H decarboxyla-
E4 ammonia E6 ketone, then enolization and althoughacidit can also OH
occur thermally, the rate increases HO OH
are monofunctional activities, whilst
(generated fromplants utilize the (2-hydroxybenzoic acid) are derived from chorismic PPO OH tive aromatization occurs with retention ofimine–enamine
the hydroxyl
CO2H H
bifunctionalCO2Hdehydroquinase–shikimate
glutamine) as dehydrogenase acid via itsCOisomer
2H isochorismic acidCO(Figure2H 4.5). some 106 -fold in the phosphoribosyl
presence of thePP enzyme. The enzyme function, though
phosphoribosylanthranilic acid as yet there is no evidence tautomerism
that any inter-
nucleophile NH2 a +
activity already mentioned. Fungi, however, possess The isomerizationOHinvolves NADan SN 2! -type ofOH reaction, achieves this by binding the pseudoaxial conformer of mediate carbonyl analogue of prephenic acid is involved.
multifunctional enzyme complex (termed AROM) that an incoming water nucleophile attacking the diene chorismic acid, allowing a transition state with chair-like Transamination of the resultant 4-hydroxyphenylpyruvic
E9 OP HO OP H
catalyses five successive
O CO2Htransformations (reactions O 2–6).CO2Hsystem and displacing OH the hydroxyl. Salicyclic OH acid geometryHO to develop. acid subsequently gives l-tyrosine. l-Arogenic
enol–keto acid is
O O OH CO2H acid OH
4-Hydroxybenzoic acid (Figure 4.5) is produced in arises by 2,3-dihydro- an elimination reaction2,3-dihydroxybenzoic
analogous to that Pathways H
H to the aromatic amino acids l-phenylalanine – CO2 the result of transamination of prephenic
tautomerism occurring
NH2 2-amino-2-deoxy- OH OH O HO
bacteria from chorismic acid by an elimination reac- and l-tyrosine via prephenic acid may vary according – H2O to
4-amino-4-deoxy- isochorismic acid producing 4-hydroxybenzoic acid from chorismic
2,3-dihydroxybenzoic acid acid, prior to the decarboxylative
OP aromatization, and canOPbe
tion, losing theacidrecently introduced enolpyruvic acid again with syn
chorismic acidstereochemistry. In the formation of the organism, and often more than None route may operate transformed OH into both l-phenylalanine and l-tyrosine OH de-
N E2 N
side-chain.
PLP The E5 stereochemistry of elimination
elimination of pyruvic acid is unusu- 2,3-dihydroxybenzoic
E7 E1: chorismate lyaseacid, the side-chain of isochorismic in a particular
H species according Hto the enzymeE3activi- pending N
H on the absence or presence of Ha suitable enzymic
ally syn, and so the mechanism is perhaps more com- acidE2:is isochorismate lost first bysynthase
hydrolysis,
(EntC) then dehydrogenation ties which are available (Figure 4.13). InPessence, only 1-(2-carboxyanilino)-
indole-3-glycerol dehydrogenase activity. In some organisms, broad activ-
CO2H CO2H
plex than at first glance. In plants, 4-hydroxybenzoic of the E3: salicylate
3-hydroxy synthase
to a 3-keto allows enolization and E4 reverse
three reactions
aldol
are involved, decarboxylative aromatiza- 1-deoxyribulose
ity enzymes5-P are known to be capable of accepting both
E4: 4-amino-4-deoxychorismate synthase (PabA, PabB) reaction
NH
acid is formed by a branch much further on in 2the formation PabAof =the aromatic
glutamine ring. 2,3-Dihydroxybenzoic
amidotransferase tion, transamination, and, in the CO2case
H of tyrosine biosyn- prephenic acid and arogenic acid as substrates. In mi-
HO
pathway via side-chain degradation of cinnamic acids acidE5:is 4-amino-4-deoxychorismate
a component of the lyase powerful
(PabC)iron chelator thesis, an oxidation, but the order in which these reactions CO2H croorganisms and plants, l-phenylalanine and l-tyrosine
E4 + E5: p-aminobenzoate NH2
(see page 157). The three phenolic acids so far en- (siderophore) enterobactinsynthase (Figure 4.6) found in occur differentiates the routes. tend to be synthesized separately, as in Figure 4.13, but
anthranilic E6: 2-amino-2-deoxyisochorismate synthase L-Ser
countered,NH4-hydroxybenzoic,
2 protocatechuic, andacidgal- Escherichia coli and many other Gram-negative bacteria.
E7: 2-amino-2-deoxyisochorismate lyase Decarboxylative aromatization NH
E4 of prephenic acid yields 2 in animals, which lack the shikimate pathway, direct
lic p-aminobenzoic
acids, demonstrate
acid some of the hydroxylation pat- SuchE6 compounds playsynthase
+ E7: anthranilate an important role in bacterial
(TrpE, TrpG) phenylpyruvic acid, and PLP-dependent transamination hydroxylation of l-phenylalanine
E1: anthranilate to l-tyrosine,
phosphoribosyltransferase (TrpD) and of
(PABA)
terns characteristic of shikimic acid-derived metabolites, growth TrpG by making available
= glutamine sufficient concentrations of
amidotransferase PLP
then leads toN H l-phenylalanine. In the presence
N of an
H
E2: phosphoribosylanthranilate
l-tyrosine to l-dihydroxyphenylalanine isomerase(l-DOPA)
(TrpC) may
i.e. a single hydroxy para to the side-chain L-Trpfunction, E8: isochorismatase
essential (EntB)comprises three molecules of
iron. Enterobactin E3: indole-3-glycerol phosphate synthase (TrpC)
E9: 2,3-dihydro-2,3-dihydroxybenzoate indole L-Trp E4: tryptophan synthase (TrpA, TrpB)
dihydroxy groups arranged ortho to each other, typi- 2,3-dihydroxybenzoic dehydrogenase (EntA)acid and three of the amino acid (enzyme-bound)
THE SHIKIMIC ACID PATHWAY CO2H
E2 elimination
of ammonia CO2H HO COSCoA
HO CO2H
HO
HO
O CO2H
NH2 E1 HO
HO HO OH OH
O OH
caffeoyl-CoA OH
chlorogenic acid
quinic acid (5-O-caffeoylquinic acid)
E1
E1: quinate O-hydroxycinnamoyltransferase
L-Phe cinnamic acid
O2 COSCoA OH OH
E3 hydroxylation sequence of hydroxylation
OH and methylation reactions
O
NADPH L-Tyr
CO2H 150 Medicinal Natural Products: A Biosynthetic
E3 Approach.
CO H3rd Edition E2CO H
CO2H HO CO2H HO2C CO2H HO2C 2 2
4-coumaroyl-CoA 4-hydroxyphenyl- 4-hydroxyphenyl-
NH2 (see page 140). Methylationlacticis catalysed
acid by a broad- ester facilitates
pyruvic acid the first reduction step by introducin
O2 specificity methyltransferase
E4 enzyme.O better leaving group (CoAS− ) for the NADPH-depen
2
NADPH Substitution
SAM patterns are not necessarily
NADPH elaborated reaction.
SAM The second reduction step, aldehyde to alco
OH OH
completelyOat theCOcinnamic
2H acid stage, and coenzyme A utilizes aOfurther COmolecule
2H of NADPH and is revers
O2
E4 esters andE5aldehydes may also 3′be substrates
E6 for aromatic TheE5enzymes involved generally exhibit rather broad
E2 NADPH HO
HO
3 hydroxylationO and methylation. Reduction
MeO MeO of cinnamic strate
OH MeO O
specificity and transform
OMe OH
substrates with the di
OH E5 E6(Figure 4.17) ent substitution patterns.
acids via coenzyme A esters and aldehydes
OH HO OH leads to the correspondingOH alcohols, precursors ofHOOH
lignans Some OH common natural cinnamic a
of the more
4-coumaroyl-4-hydroxyphenyl- rosmarinic acid
L-Tyr 4-coumaric acid and
caffeic acid lignin (see
lactic page 151). Formation
ferulic acid
acid of the coenzyme A are
5-hydroxyferulic 4-coumaric, caffeic,
sinapic acid ferulic, and sinapic a
(p-coumaric acid) acid
E2: tyrosine aminotransferase E4: hydroxycinnamoyl-CoA:hydroxyphenyllactate
E3: hydroxyphenylpyruvate reductase hydroxycinnamoyl
HO CO2H transferase (rosmarinic acid synthase)
HO
E5, E6: hydroxycinnamoyl-hydroxyphenyllactate 3- and 3′-hydroxylases
HO COSCoA
HO
O CO2H
CH2OH CH2OH E1 HO CH2OH
HO HO OH OH
sinapic acid O OH
caffeoyl-CoA OH
E7 UDPGlc OMe chlorogenic acid
OMe
OH quinic acid OH (5-O-caffeoylquinic acid)
OH Me 3N OH
O E1: quinate O-hydroxycinnamoyltransferasecholine
HO
HO O O
OMe Me3N OMe
OH MeO MeO OMe
O COSCoA E8 OH O OH
OH O
OH 1-O-sinapoylglucose OH glucose OH
sinapine L-Tyr
HO HO2C E3 E2
4-hydroxycinnamyl alcohol coniferyl alcohol
HO2C
sinapyl alcohol
(p-coumaryl alcohol)E7:
sinapate 1-glucosyltransferase
4-coumaroyl-CoA 4-hydroxyphenyl- 4-hydroxyphenyl-
lactic
E8: sinapoylglucose:choline O-sinapoyltransferase (sinapine acid
synthase) pyruvic acid
E4
Figure 4.18 OH OH
E1: phenylalanine ammonia lyase (PAL) O CO2H
O2
O CO2H
E2: tyrosine ammonia lyase (TAL) POLYMERS 3′ HO
NADPH
E3: cinnamate 4-hydroxylase ×2 3 O ×n O OH
E4: p-coumarate 3-hydroxylase E5 E6
HO HO
E5: caffeic acid O-methyltransferase 4-coumaroyl-4-hydroxyphenyl- rosmarinic acid
LIGNANS lactic acid LIGNIN
 H H
HO HO

THE SHIKIMIC ACID PATHWAY

152 Medicinal Natural Products: A Biosynthetic Approach. 3rd Edition
OH
these two steps probably involve
OH ring opening to the quinonemethide OH
phenolic oxidative followed by reduction
OH OH OH OH OH coupling O O
one-electron OMe OMe
oxidation H H NADPH H H
MeO MeO
E1 E2 HO
– H+ O
MeO
–e OH HO HO (+)-lariciresinol
MeO MeO MeO MeO MeO (+)-pinoresinol
coniferyl alcohol
OH O O O O
E2 NADPH
coniferyl alcohol A B C D oxidation to CHO, then
resonance forms of radical hemiacetal (lactol) formation OH
MeO MeO
OH OH
A + D B + D D + D O OMe
radical radical radical NAD+ OH
HO HO H H
pairing pairing pairing
OH ≡ MeO
OH OH E3 HO
O H
HO
MeO MeO
OMe (–)-secoisolariciresinol
OH OH
HO oxidation
H OH to lactone (–)-secoisolariciresinol
E3 NAD+
HO OMe OMe
O O MeO O O
enolization HO O O O
H2O O O O nucleophilic
OH nucleophilic attack HO
attack onto
OMe MeO of hydroxyls onto O O O
O H quinonemethide
quinonemethides modification
O system
OMe nucleophilic attack H of aromatic
of water onto MeO substitution MeO OMe MeO OMe
quinonemethide HO patterns
OH OH OMe OMe
OH OMe matairesinol yatein
O
OH OMe
OH hydroxylation hydroxylation OH
H MeO
O O O2 O O2 O
O
OMe OH O NADPH O NADPH O
nucleophilic attack HO O O O
HO OMe pinoresinol E4 E5
of hydroxyl onto O O O
O quinonemethide (resinol linkage)

HO
HO MeO OMe MeO OMe MeO OMe
OMe OMe OMe OMe
OH
O podophyllotoxin deoxypodophyllotoxin β-peltatin
OMe
guaiacylglycerol E1: pinoresinol synthase E4: deoxypodophyllotoxin 7-hydroxylase
β-coniferyl ether HO dehydrodiconiferyl alcohol E2: pinoresinol-lariciresinol reductase E5: deoxypodophyllotoxin 6-hydroxylase
(β-arylether linkage) OMe (phenylcoumaran linkage) E3: secoisolariciresinol dehydrogenase

Figure 4.19 Figure 4.20

 in the oil from the bark of cinnamon (Cinnamomum zey- allylphenol found in flavouring materials. Myristicin also 6 3

lanicum; Lauraceae), widely used be as areduced

spice and Gaultheria procumbens (Ericaceae), used for many years The two 2-hydroxycinnamic acids then suffer a change in
by flavour-
addition ofhas hydride
a history(from of NADPH)
being employedgen- as a mild hallucinogen
THE SHIKIMIC ACID PATHWAY
ing. Fresh bark is known to containerating
nce from an appropriate cinnamic acid
high levels of the ester
the different side-chains according
via ingestion of toground
the position
nutmeg. It islation
for pain relief. Ithydroxylation
probably COmetab-
2H
of salicylic
is derived by SAM-dependent
acid.
O2 The salicyl
CO2H
methylation
methy- configuration in the side-chain, from the trans (E) to the
alcohol derivative less
CO2Me CO H
stable cis (Z) form. Whilst trans–cis 2isomerization
cinnamyl acetate, and cinnamaldehyde is released
of attack; the genes fromand olized
enzymes in theinvolved
body via in the two
an amination reaction
salicin,157 to giveinanmany
found NADPH species of willow OH (Salix SAMspecies; would OH OCOCH3
g cinnamyl alcohol is utilized for the The Shikimate Pathway: Aromatic Amino Acids and Phenylpropanoids be unfavourable in the case of a single isolated
this by fermentation processes which processes haveof
are part been
com- characterized.
amfetamine-like derivative (see page 404). Anethole
Salicaceae), is notis derived from salicylic acid, but prob- double bond, the fully conjugated system in the cinnamic
ous phenylpropene derivatives. Thus E1 E2
mercial preparation of the bark, presumably Myristicin (Figure 4.24)
by enzymic the main from component
nutmeg (Myristica
in oils from aniseed ably via(Pimpinella
glucosylation of salicylaldehyde and then reduc- acids allows this process to occur quite readily, and UV
Figure 4.24) is the principal component fragrans; Myristicaceae) anisum; is a further attack of
example of an star anise
Umbelliferae/Apiaceae), (Illicium ver- (Figure salicylic
hydrolysis and participation of the reversible alcoholleaving
loss of acetate de- hydride at tion of benzoic acid
the carbonyl acid
4.28). Salicin methyl salicylate
is responsible aspirin equilib-
irradiation, e.g. daylight, is sufficient to produce
bark of cinnamon (Cinnamomum zey- allylphenol group; found
formation of
in flavouring H
materials.
um; Illiciaceae) Myristicin
and fennel also (Foeniculumforvulgare; Umbel- (acetylsalicylic acid)
hydrogenase. Cinnamon leaf, on the other hand, contains g-position the analgesic and antipyretic effects of willow barks, rium mixtures which can be separated. The absorption of
), widely used as a spice and flavour- has a quinonemethide history side-chain
large amounts of eugenol (Figure 4.24) and muchofsmaller
being employed as a mild hallucinogen
liferae/Apiaceae). The presence of propenyl widelycomponents
used for centuries. It provided the template for energy promotes an electron from the π-orbital to a higher
cleavage
nown to contain high levels ofOthe ester via ingestion
is also theofprinci-
ground nutmeg. It is probably
materials metab-
Me O Me in flavouring such as cinnamon, synthesisstar anise,
of acetylsalicylic acid (aspirin; Figure 4.28), a energy state, i.e. the π∗ -orbital, thus temporarily destroy-
amounts of cinnamaldehyde. Eugenol
and cinnamaldehyde is released from olized in the body via an amination reaction to give
nutmeg, and sassafras (Sassafras albidum; an CO
Lauraceae)
more effective H CO2H
and widely used pain-killer.
2 ing the double-bond character and allowing rotation. Loss
pal OH
constituent in oil from
esterification O cloves (Syzygium O aromaticum;
n processes which are part of com- amfetamine-like derivativehas (see page 404).
reduced Anethole isuse somewhat, since these
their commercial side-chain of the absorbed energyCHOthen resultsreduction
in re-formation of the
Myrtaceae), used for many years as a dental anaesthetic OMe OMe
hydroxylation CHO glucosylation CH2OH
of the bark, presumably by enzymic the main component in oils from aniseed
constituents have been(Pimpinella
shown to be weak carcinogens cleavage double bond, but in the cis-configuration. In conjugated
as well as for flavouring. OH OH UDPGlc OGlc OGlc
icipation of the reversible alcohol de- anisum; Umbelliferae/Apiaceae),
NADPH OHin laboratory star anisetests(Illicium
OH ver- In the case of safrole
on animals. systems, the π–π∗ energy difference is considerably
Allylphenols
CH3COSCoA (e.g. eugenol) or propenylphenols (e.g. isoeugenol Coumarins
mon leaf, on the other hand, contains um; Illiciaceae) E1 and fennel(Figure
(Foeniculum mainUmbel-
vulgare;
4.26),(propenylphenol)
the component of sassafras oil, this less than with a non-conjugated double bond. Chemical
isoeugenol) both originate from
genol (Figure 4.24) and much smaller
≡ appropriate cinnamyl
liferae/Apiaceae). The al- presence
toxicity of propenyl
has been components
shown to arise from
The hydroxylation
hydroxylation of cinnamic acids ortho to the lactonization can occur on treatment with acid. Both the
cohols
OMeby way of acetateOMe esters (Figure 4.25),
OMe though NADPHthe such cinnamic acid 2-coumaric acid of salicylicsalicylaldehyde
aldehyde. Eugenol is also the princi- in flavouring materials in theas cinnamon,
side-chain
attack of hydride star anise,
followed by sulfation,
side-chain giving
as an
seen in the biosynthesis acid trans–cis isomerization and the lactonization salicin are enzyme
OH two groups of compounds OH differ with OHrespect
nutmeg, andto the
E2 posi-
sassafras (Sassafras at albidum; Lauraceae)
agent which binds to cellular macromolecules.
a-position (Figure 4.28) is
Furthera crucial step in the formation of a group mediated in nature, and light is not necessary for coumarin
il from cloves (Syzygium aromaticum; E1: benzoic acid 2-hydroxylase
tion
coniferyl of the side-chain double
coniferyl bond. The acetate function
has reduced their commercial data useH on somewhat,
volatile oils since these
containing of cinnamic
aromatic constituents
E2: acidacid
salicylic lactone
carboxylderivatives, the coumarins. biosynthesis. However, the enzyme activities are poorly
methyltransferase
or manyalcohol
years as a dental anaesthetic SAM
provides a leavingacetate group, loss of constituents
which is facilitated
have been by shown
isolated to from
be weak thesecarcinogens
and other plant materials Whilst theare direct
given hydroxylation of the aromatic ring of the characterized. Cinnamic acid and 4-coumaric acid give
uring.
the presence of the para hydroxylingroup laboratory tests on animals.
on the aromatic in TableIn4.1. theVolatile
case ofoils safrole
in which
E3 the cinnamic
Figure
main OMe 4.28 is common, hydroxylation generally in- rise to the coumarins coumarin and umbelliferone
acids
components
g. eugenol) E1: or isoeugenol
propenylphenols (e.g. OMe OMe volves initially the 4-position para to the side-chain, and respectively (Figure 4.29). Other coumarins with
synthase 1 (Figure 4.26), the main component of
ring, leading to a quinonemethide intermediate. This can are terpenoid in nature are listed in Table sassafras oil, this OMe 5.1, page 158.
iginate fromE2:appropriate cinnamyl
eugenol synthase 1 al- toxicity has been shown OH to arise from hydroxylation OH
cetate estersE3: eugenol O-methyltransferase
(Figure 4.25), though the in the side-chain followed by sulfation, eugenol
giving an
(allylphenol) CO2H CO2H
pounds differ with respect to the posi- agent which binds to cellular macromolecules. Further
ainFigure
double4.25 bond. The OCOCH3
CHOacetate function data on volatile oils containing aromatic constituents E2
CO2H
group, loss of which is facilitated by isolated from these and other plant materials are given OH OH O O
cinnamic acid 2-coumaric acid coumarin
para hydroxyl group on the aromatic in Table 4.1. Volatile oils in which the main components trans–cis lactone
giveare vanillic acid (4-hydroxy-3-methoxybenzoic acid)O2 isomerization formation
uinonemethide intermediate. This can are terpenoid in nature listed in Table 5.1, page 158. NADPH
E1
(Figure 4.27). Alternatively, cinnamic acid itself may be
HO HO3SO CO2H CO2H
converted into benzoic acid. A sequence analogous to that
MeO MeO MeO
O (see OMe
involved in the β-oxidation of fatty acids page 18) CO2H
OCOCH
cinnamaldehyde cinnamyl OMe OMe OHthe double bond O in the coenzyme E2 HO
3 is possible, so that HO OMe A OH HO OH HO O O
acetate
anethole ester would be eugenol
estragole hydrated, the myristicin
hydroxyl group oxidized elemicin to
4-coumaric acid 2,4-dihydroxy- umbelliferone
O O O (methylchavicol)
a ketone, and the β-ketoester would then lose acetyl-CoA cinnamic acid
O O O by a reverse Claisen reaction, giving the coenzyme A es-
Figure 4.24
safrole safrole 1′-sulfate ter of 4-hydroxybenzoic acid. Whilst this sequence has MeO MeO HO
been generally accepted, there is also evidence to support
Figure 4.26 MeO MeOanother side-chain O MeO cleavage mechanismOMe which is differ- GlcO O O HO O O HO O O
cinnamyl OMe OMe OH O OMe
ent from the fatty acid β-oxidation pathway (Figure 4.27).
acetate scopolin scopoletin aesculetin
anethole estragole eugenol Coenzyme
myristicinA esters areelemicinnot involved, and though E1: E2:
cinnamate 4-hydroxylase
a similar
cinnamate/coumarate 2-hydroxylase
(methylchavicol) hydration of the double bond occurs, chain shortening fea-
THE MIXED SHIKIMIC ACID-ACETATE PATHWAY
The Shikimate Pathway: Aromatic Amino Acids and Phenylpropanoids 169 The Shikimate Pathway: Aromatic Amino Acids and Phenylpropanoids 171

chain extension; acetate pathway OH OH OH

with a cinnamoyl-CoA starter group
OH 3 × malonyl-CoA OH HO O HO O HO O
R O2 R O2 R
2-oxoglutarate 2-oxoglutarate
CoAS CoAS
E1 OH E4 OH
O O O O E3
E2 O OH O OH O OH O
4-hydroxycinnamoyl-CoA
R = H, naringenin R = H, dihydrokaempferol R = H, kaempferol
different folding R = OH, eriodictyol R = OH, dihydroquercetin (taxifolin) R = OH, quercetin
modes (flavanones) (dihydroflavonols) (flavonols)
OH OH OH O2 O2
2-oxo- E1 E2 NADPH E5 NADPH
O SCoA O SCoA glutarate
O
OH OH OH
NADPH
O O HO
O
SCoA HO O HO O O2 HO O
E3 R R R
O O OH O 2-oxoglutarate
O O
OH OH
E1 Claisen E1 Claisen E6
E2 aldol + OH O OH OH OH OH
ester hydrolysis OH OH
OH R = H, apigenin R = H, leucopelargonidin
O O O O R = OH, luteolin R = OH, leucocyanidin
OH NADPH (flavones) NADPH (flavandiols; leucoanthocyanidins) – 2 H2O
O
E3 E8
H H OH OH OH
CO2H O O OH O
O dehydration HO O HO O HO O
decarboxylation enolization enolization R R NADPH R
dehydration (E1) (E1)
OH OH
enolization OH OH E7 OH
CO2 E2
OH HO OH HO OH OH OH OH

HO R = H, (+)-afzelechin R = H, (−)-epiafzelechin R = H, pelargonidin

R = OH, (+)-catechin R = OH, (−)-epicatechin R = OH, cyanidin
(2,3-trans-flavan-3-ols; catechins) (2,3-cis-flavan-3-ols; catechins) (anthocyanidins)
OH O O
naringenin-chalcone isoliquiritigenin
OH (chalcone) Michael-type nucleophilic (chalcone) E1: flavone synthase I E5: dihydroflavanol 4-reductase
resveratrol attack of OH onto E2: flavone synthase II E6: anthocyanidin synthase (leucoanthocyanidin dioxygenase)
(stilbene) a,b-unsaturated ketone E3: flavanone 3-hydroxylase E7: anthocyanidin reductase
E4 E4: flavonol synthase E8: leucoanthocyanidin reductase
E4
OH OH
E1: chalcone synthase Figure 4.43
(naringenin-chalcone synthase) HO O HO O
E2: stilbene synthase (resveratrol synthase)
E3: chalcone reductase is growing belief that some flavonoids are particularly colours: yellows from chalcones and flavonols, and reds,
E4: chalcone isomerase
beneficial, acting as antioxidants and giving protection blues, and violets from anthocyanidins. Even the colour-
OH O O
naringenin liquiritigenin against cardiovascular disease, certain forms of cancer, less materials, e.g. flavones, absorb strongly in the UV and
(flavanone) (flavanone) and, it is claimed, age-related degeneration of cell compo- are detectable by insects, probably aiding flower pollina-
 OH
OH OH
THE MIXED SHIKIMIC ACID-ACETATE PATHWAY
OH HO
172 Medicinal Natural Products: A Biosynthetic Approach. 3rd Edition
O
OH
O
OH
one-electron
oxidation to radical O
one-electron
oxidation
OH HO OH HO O HO O
OH – H+ OH
OHOH OH OH – H+
HO O
OH OH OH –e OH
HOO O –e
OH HO HO O OMe OMe
OH OH OH O OH O
O OH HO
O OH O OH
OH taxifolin
OH OH OH OH OH
O OH coniferyl alcohol
OH OH radical
HO O O OH O coupling
OH OH OH
HO O OH
HO O OH HO O
O OH OH HO O OMe OH
OH O
OH OHOH OH OH O HO OH O
O OH OH
epigallocatechin gallate OH theaflavin OH OH
OH OH HO O OMe
HO O OH O OH
OH OH
epicatechin trimer silybin A
OH HO OH OH
OH OH O H
epigallocatechin gallate theaflavin O
Figure 4.44 OH OH O
nucleophilic attack of OH
epicatechin trimer HO O OMe onto quinonemethide (alternative
O stereochemistries possible)
OH
Figure 4.44 HO OH OH
HO OH O
O silybin B The Shikimate Pathway: Aromatic Amino Acids and Phenylpropanoids 175
OH
HO
L-Rha (diastereoisomeric pair = silybin)
O OMe OH
HO oxidation 1,2-aryl
O OH OH
HO O Figure 4.47 to radical migration
HO O O HO O O2 OH
L-Rha OMe OH OH OH HO O HO O HO O
O NADPH
HO O OH a remarkable change to their taste, and the products are radical generated from coniferyl alcohol (Figure 4.47).
Fe Enz
D-Glc O O HO O O−Glc−Rha H E1
HO OH OH now intensely sweet, being some 300–1000 times as This would lead to an adduct which could cyclize by
H O
rutinose OH OH O OH O sweet as sucrose. Neohesperidin-dihydrochalcone is usedR attack
R O
Fe Enz O of the phenol nucleophile ontoR the O quinone me-
= rhamnosyl(α1→6)glucose
D-Glc hesperetin O−Glc−Rha
quercetin rutinose as a non-sugar sweetening agent. thide system provided by coniferyl alcohol. The product OH
R = H, liquiritigenin
rutinose OH O OH O R = OH, naringenin would be silybin, found in Silybum marianum (Composi-
= rhamnosyl(α1→6)glucosehesperidinhesperetin quercetin rutin
rutinose Flavonolignans (flavonoids) tae/Asteraceae) as a mixture HO of two trans O diastereoiso-
HO 7 O OH
An interesting combination of flavonoid and lignan-like mers, silybins A and –B,
H 2reflecting
O a lack of stereospeci-
hesperidin
OH OMe rutin
structures is found in a group of compounds called ficity for the original radical coupling. In addition, the
O E3
HO
OH OMe flavonolignans. They arise by oxidative coupling pro- regioisomer isosilybin (Figure 4.48), again a mixture of
D-Glc HO O O OH R O
trans
4′
diastereoisomers, is also found inR Silybum.
O In keep-
O OH cessesE1:
between a flavonoid and a
2-hydroxyisoflavanone synthase
phenylpropanoid, the OH OH
HO OO latter E2:
usually ing with
coniferyl alcohol. Thus, the dihydroflavonol R = H, daidzeinthe postulated biosynthetic origin, mixtures of
D-Glc HO O OH 2,7,4′-trihydroxyisoflavanone
OH Rha−Glc−O O taxifolin (dihydroquercetin)
4′-O-methyltransferasethrough one-electron oxida-R = silybin
OH, genistein
and isosilybin may be obtained byE2 incubating
SAM tax-
OO E3, E4:
tion may 2-hydroxyisoflavanone
provide combine with the (isoflavonoids)
dehydratase
a radical, which may ifolin and coniferyl alcohol with a peroxidase preparation
HO OH O Rha−Glc−O O
O HO O HO O OH
HO
L-Rha HO OH O – H 2O
OH
L-Rha HO OH O
neohesperidose OH E4
= rhamnosyl(α1→2)glucose OH O naringenin
neohesperidose hesperetin neohesperidose
R O R O
= rhamnosyl(α1→2)glucose hesperetin neohesperidose naringenin OMe OMe
neohesperidin naringin R = H, formononetin
neohesperidin naringin R = OH, biochanin A
(isoflavonoids)
Figure 4.45
 O O
ubiquinone-n plastoquinone-n
(coenzyme Qn) R1 = R2 = Me, α-tocopherol
THE MIXED SHIKIMIC ACID-TERPENES
The Shikimate Pathway: Aromatic Amino AcidsPATHWAY
and Phenylpropanoids 179 O O R1 = H, R2 = Me, β-tocopherol
n = 1–13 R1 = Me, R2 = H, γ-tocopherol
R1 = R2 = H, δ-tocopherol
O O R2
(vitamin E)
MeO n = 1–12 n = 3–10 HO H H
3 n
O O
H phylloquinone menaquinone-n
MeO H H R1 O
n n (vitamin K1; phytomenadione) (vitamin K2)
O O 3
ubiquinone-n plastoquinone-n
Figure 4.53
(coenzyme Qn) R1 = R2 = Me, α-tocopherol
O O R1 = H, R2 = Me, β-tocopherol
n = 1–13 R1 = Me, R2 = H, γ-tocopherol CO2H
R1 = R2 = H, δ-tocopherol
(vitamin E)
H H H DMAPP + IPP
3 n OPP E2
O O plants/ n
phylloquinone menaquinone-n animals CO2H CO2H
(vitamin K1; phytomenadione) (vitamin K2)
E3
OH
Figure 4.53 4-coumaric acid C-alkylation with a
bacteria polyisoprenyl PP nH
CO2H OH OH
E1 4-hydroxybenzoic
CO2H
CH3COCO2H acid in eukaryotes: in prokaryotes:
1. hydroxylation O2 1. decarboxylation E5
DMAPP + IPP 2. O-methylation E4 SAM 2. hydroxylation E6
H O CO2H
OPP E2 3. decarboxylation 3. O-methylation E7
plants/ n
animals CO2H CO2H OH
chorismic acid
E3
OH MeO
4-coumaric acid C-alkylation with a nH
bacteria polyisoprenyl PP nH OH
CO2H OH OH 1. C-methylation
E1 4-hydroxybenzoic 2. hydroxylation E8 O2 oxidation
CH3COCO2H acid in eukaryotes: in prokaryotes: O 3. O-methylation O to quinone
1. hydroxylation O2 1. decarboxylation E5
2. O-methylation E4 SAM 2. hydroxylation E6 MeO SAM O2 SAM
O CO2H 3. decarboxylation 3. O-methylation E7
OH MeO E4 E10 E9 MeO
chorismic acid nH E7 n
H
O O
ubiquinone-n
MeO
nH
OH E1: chorismate lyase (UbiC) E6: UbiB Coq: Saccharomyces cerevisiae (n = 6)
1. C-methylation E2: polyprenyl diphosphate synthase (Coq1, IspB) E7: UbiG Ubi, Isp: Escherichia coli (n = 8)
2. hydroxylation E8 O2 oxidation E3: 4-hydroxybenzoate polyprenyltransferase (Coq2, UbiA) E8: Coq6, UbiH
O 3. O-methylation O to quinone E4: Coq3 E9: Coq5, UbiE
E5: UbiD E10: Coq7, UbiF
MeO SAM O2 SAM