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Shikimates and
Phenylpropanoids
RA Macahig
FM Dayrit
OH
CO2H
HO
HO
OH
OH
OH
OH
shikimic acid
luteolin
Introduction
Shikimic acid is the key intermediate of a large group
aromatic natural products. The isolation of shikimic acid was
first reported from aniseed (Illicium anisatum) and the fruit of
I. religiosum, whose Japanese name was shikimi-no-ki (shi
four; kimi seasons; no of; ki tree, literally tree of four
seasons). Shikimic acid has since been found in many plants,
bacteria, yeasts and moulds.
It is estimated that this group accounts for 35%
CO2H
of plant dry mass, and that one-fifth of the
carbon fixed by plants is channeled to shikimic
acid metabolites, such as the lignins.
Shikimic acid is the precursor of three important
aromatic amino acids: phenylalanine, tyrosine,
and tryptophan.
8.0 Shikimates & phenylpropanoids (Dayrit)
HO
OH
OH
shikimic acid
2
Overview
OP
D-glucose
H
pentose
phosphate
cycle
H+
_
O2C
phosphoenol pyruvate
glycolysis
OH
PO
H-O
O
H
_
O2C
HO
OH
OH
heptulose
D-erythrose-4-phosphate
H H
_
CO2
HO
_
CO2
OH
OH
OH
OH
5-dehydroquinate
3-dehydroshikimate
_
CO2
HO
OH
OH
shikimate
HOCH2
H
HO
O
H
OH
OH H H
H
OH
D-glucose
CH3
CO2H
O
CO2H
1
CO2H
EPSPS
HO
OH
OH
shikimic acid
CO2H
PO
benzoic acids,
tryptophan
OH
5-enolpyruvylshikimate
-3-phosphate
CO2H
CO2H
OH
chorismic acid
EPSPS: 5-enolpyruvyl
shikimate-3-phosphate
synthase
phenylalanine,
tyrosine
O
HO2C
lignans,
lignins
CO2H
phenyl propanoids
+ (3 x Ac-CoA)
phenols
OH
prephenic acid
flavonoids
styrenes
benzoic acids
In microorganisms:
[O]
polyketide
aromatic compound
quinone
In plants:
[O]
Overview of
biosynthesis of
quinones. Depending
on the organism,
quinones can arise via
the polyketide or
shikimate pathways.
polyketide
aromatic compound
quinone
quinone
shikimate
aromatic compound
+ terpene
[O]
quinone
(mixed metabolite)
quinone from shikimate:
OH
OH
OH
CO2H
OH
homogentisic acid
alkarinin
O
R
5. Polyketides (Dayrit)
CO2H
NH2
phenylalanine, R=H
tyrosine, R=OH
O
cinnamic acid
coumarin
CO2H
O
CO2H
phenylacetic acid
benzoic acid
O
flavonoid
O
O
lignan
_
CO2
CO2
H
H
HO
1. ATP
OH
OH
2. PEP
shikimate
PO
_
CO 2
_
CO2
OH
3
2
CO2
OH
HO
[3,3]sigmatropic
shift
_
CO2
O2C
_
CO2
CO2
3
3
1
_
CO2
OH
chorismate
5-EPSP
_
O2C
PO
OH
_
CO2
OH
p-hydroxybenzoic acid
prephenate
o-aminobenzoic acid,
p-aminobenzoic acid
phenylpropanoids
CO2
Prephenate is
the precursor of
phenylalanine
and tyrosine.
NH2
_
CO2
CO2H
O
a
-CO2,
-H2O
Pyridoxamine
transaminase
OH
phenylalanine
prephenate
b [O]
_
O2C
CO2
NH2
_
CO2
CO2H
O
-CO2
O
Pyridoxamine
transaminase
OH
OH
tyrosine
Routes to benzoquinones:
A. Polyketides
Hydroxybenzoic acids
B. Shikimatic acid
Benzoquinones
Homogentisic acid
C6 Metabolites
Benzoquinones
Chroismic acid
p-Hydroxybenzoic acid
Phenylalanine
Benzoquinones
CO 2H
R=
1. -CO 2
2. [O]
3. O-methylation
from terpenes
MeO
OH
OH
Me
Me
MeO
MeO
O
Ubiquinone
R
O
5-Demethoxy
ubiquinone
[O]
Quinones are
formed mainly from
the polyketide and
shikimate pathways,
although other
groups may produce
quinones from
extensive
modification. The
biosynthesis of
R
OH
ubiquinone, the
1. [O]
compound which
2. C-methylation
assists in biological
electron transfer, is
OH
Me
shown.
MeO
R
OH
10
C6-C1. Tannins are a group of benzoic acid plant metabolites which precipitate
proteins. The gallotanins, which are hydrolyzable tannins, contain glucose and
gallic acid joined by ester linkages.
HO
O
HO
OH
HO
HO
OH
OO
HO
OH
corilagin
OH
HO
HO
OH
OH
OH
O
O
OH
OH
O
HO
O
HO
OH
O
O
HO
OH
OH
O
O
HO
O
O
OH
O
HO
HO
OH
O
OH
Turkish tannin
HO
OH
11
Two benzoic acid derivatives of some interest are gallic acid and ellagic acid.
Gallic acid and ellagic acid are constituents of hydrolyzable tannins but these are
found in many other natural products. These compounds are natural antioxidant
in aqueous and micellar environments. Ellagic acid is also a naturally-occuring
phytochemical pesticide and antimicrobial.
OH
OH
HO
O
OH
HO
HO
HO
OH
OH
O
O
O
HO
HO
O
O
Gallic acid
OH
HO
OH
HO
OH
Ellagic acid
12
OH
HO
OH
OH
OH
R1
HO
HO
HO
HO
R2
OH
O
OH
OH
OH
CH2
O
HO
O
HO
HO
O
O
O
O
OH
OH
13
-o xidation
PAL
The benzoic
and cinnamic
acids are
biosynthesized
in a metabolic
grid. These
compounds
occur widely
in plants.
CO 2H
CO 2H
CO 2H
NH2
cinnamic acid
phenylalanine
[O]
[O]
CO 2H
NH2
HO
be nzoic acid
[O]
CO 2H
TAL
HO
p-hydroxybenzoic acid
[O]
[O]
HO
CO 2H
HO
[O]
CO 2H
-o xidation
NH2
HO
HO
HO
CO 2H
de carbo xylase,
-CO2
CO 2H
OH
gallic acid
[CH3]
CH3O
HO
HO
HO
caffe ic acid
3,4-dihydroxyphenylalanine (DOPA)
HO
chorismic
acid
HO
4-hydroxycinnamic acid
(p-coumaric acid)
tyrosine
CO 2H
-o xidation
[CH3]
CO 2H
CH3O
CO 2H
-o xidation
NH2
HO
HO
HO
ferulic acid
do pamine
vanillic acid
1. [O]
2. [Me]
1. [O]
2. [Me]
1. [O]
2. [Me]
OH
CH3O
HO
CH3O
CO 2H
CO 2H
-o xidation
NH(CH3)
HO
adrenaline
HO
HO
OCH3
sinapic acid
OCH3
syring ic ac id
14
15
Glucosinolates
OH
HO
HO
OSO3
S
HO
HO
(R group
variable)
16
CO2H
CO2H
PAL
or
TAL
Ar - C3: cinnamaldehydes
cinnamyl alcohols
aryl propanes
Ar - C3: coumarins
C4H
R
CO2H
R1
R3
phenylalanine, R=H
tyrosine, R=OH
R2
+ 3 x Ac
cinnamic acids
Ar - C2: styrenes
acetophenones
flavonoids
Ar: phenols
quinones
18
HO CO2H
Examples of
cinnamic acids
and the C6-C3
derivatives.
Cinnamic acids
themselves do
not usually
occur in the
free form but
are isolated as
glycosides.
Cinnamic acids
are precursors
of lignans and
lignins.
O
HO
OH
O
OH
OH
OH
HO
O O
OH
HO
HO
OC
OH
CH3O
OH
RO
coniferyl alcohol, R=H
coniferin, R=glucosyl
O
2-caffeoylarbutin
CH3O
eugenol
Eugenol and saffrole are well-known
constituents of flavor and spice plants.
They are precursors to lignins and lignans.
HO
O
saffrole
O
19
HO
CO2H
NH 2
HO
HO
tyrosine
OH
O
caffeic acid
CH2 CH CO2H
HO
OH
HO
HO
CO2H
CO2H
rosmarinic acid
Rosmarinic acid is a naturally-occurring
anti-histamine compound from
tsaang gubat (Ehretia macrophylla).
OH
NH 2
HO
phenylalanine
CO2H
CO2H
glucose
RO
O-glu
RO
OH
E
umbelliferone
herniarin
skimmin
aesculetin
scopoletin
R1
R2
H
H
CH3 H
glu
H
H
OH 8.0
CH3 OCH3
R2
6
7
3
2
1
8
Shikimates
O (Dayrit)
R1O & phenylpropanoids
O
RO
CO2H
O-glu
20
O-H
CO2H
H
OH
HO
O-H
CH2
dicoumarol
HO
OH
O
O
O
HH
21
oxidase only
OCH3
OH
OH
O
CH3O
HO
CH 3O
OH
OH
OCH3
OCH3
OH
H
(+/-)-Dehydrodiconiferyl alcohols
+
OCH3
OCH3
OH
OH
O
O
H
H
O
O
HO
HO
OCH3
(+/-)-Pinoresinols
OCH3
(+)-Pinoresinol
+ Other dimers
8.0 Shikimates
&etphenylpropanoids
(Dayrit)
(N.G. Lewis,
al. in Science, 275, 362 (1997);
22
CH2OH
CH 2OH
.
[O]
CH 3O
CH3O
O.
OH
CH3O
CH2OH
CH2OH
CH3O
O
OCH3
O
H+
O
HO
OCH3
HO
O
[H]
OCH 3
CH 3O
CH 2OH
HO
CH 2OH
OH
OCH 3
O
H+
OCH3
OCH 3
HO
(+)pinoresinol
O
H+
CH 3O
CH2OH
HO
CH2OH
OCH3
OH
(+)isolariciresinol
23
.
CH3O
.
CH3O
O
O
OCH3
CH3O
O
OCH3
H+
O
OCH3
HO
licarin A
(from eugenol)
24
Lignins
The majority of carbon in the
phenylpropanoid group is channeled
toward the synthesis of lignin, a
complex three-dimensional polymer
that is a principal structural
component of plant cell walls.
(/www.scielo.cl)
Lignin has far-reaching impacts on agriculture, industry and
the environment, making phenylpropanoid metabolism a
globally important part of plant biochemistry.
Lignin is the second most abundant polymer on earth, next to
cellulose. Lignin is a major carbon sink in the biosphere,
accounting for about 30% of the more than 1.4 1012 kg of
carbon sequestered into terrestrial plant material each year.
8.0 Shikimates & phenylpropanoids (Dayrit)
25
OH
CH3O
O
HO
OCH3
O
Ph. + Ph.
O
OCH3
alkyl. + alkyl.
O. + alkyl.
HO
O
O
OCH3
OH
Ph. + alkyl.
HO
OCH3
OH
HO
Lignins are
extensively
crosslinked
phenylpropanoid
polymers. Similar to
the lignans, they are
formed by coupling
between different
radicals arising from
phenylpropanoids.
Lignins contribute to
the strength and
robustness of plants
and as a protective
barrier against
biochemical
degradation by
microorganisms.
CH3O
O
26
8.0 Shikimates
& phenylpropanoids (Dayrit)
27
Humphreys and Chapelle, Current Opinion in Plant Biology 2002, 5:224229.
Flavonoids
The flavonoids are a very widely occurring group of natural
products. There are over 5,000 flavonoids identified in plants.
Most flavonoids, except for the catechins, are naturally found
in their glycosylated form. Flavonoids are universally
distributed among vascular plants and are found throughout
the plant- roots, bark, fruit, seeds, leaves and flowerswhere
they play important functions.
Flavonoids are physiologically important because of their
many antioxidant properties.
Flavonoids have a characteristic C6-C3C6 structure, with three rings: an aromatic
A and B ring, and a -pyrone C ring.
8.0 Shikimates & phenylpropanoids (Dayrit)
28
OH
Flavonoids
HO
OH
OH
luteolin
29
OH
Flavonoids
HO
OH
OH
luteolin
30
R
1. PAL / TAL
2. CoA ligase
CoA-S
-O C
2
NH2
CO2-
phenylalanine, R=H
tyrosine, R=OH
1.
CoA -S
2. -CO2, -CoASH
R
1. NADPH
2. -CO2, -CoASH
O
benzal acetone
CoA-S
R
O
-CoASH
CO2-
1.
OH
R
CoA-S
*
CoA -S
2. -CO2, -CoASH
dihydropyrone
R
O
Overview of the
condensation of
phenylpropanoid
+ (3 x acetyl
CoA).
O O
-CoASH
O
*
styryl pyrone
CO2-
1.
CoA -S
2. -CO2, -CoASH
R
R
OH
HO
to the
flavonoids
-CoASH
*
O
O
chalcone
S-CoA
*
O
*
OH
chalcone
synthase
31
R
1. PAL / TAL
2. CoA ligase
CoA-S
-O C
2
NH2
CO2-
phenylalanine, R=H
tyrosine, R=OH
1.
CoA -S
2. -CO2, -CoASH
R
1. NADPH
2. -CO2, -CoASH
O
benzal acetone
CoA-S
R
*
O
CO2-
1.
CoA-S
-CoASH
O O
*
O
(C6-C3) + C6.
R
*
O
CoA -S
2. -CO2, -CoASH
dihydropyrone
R
O
(C6-C3) + C4;
OH
(C6-C3) + C2;
-CoASH
styryl pyrone
CO2-
32
2. -CO2, -CoASH
O
benzal acetone
CoA-S
R
O
-CoASH
CO2-
1.
OH
R
CoA-S
*
2. -CO2, -CoASH
dihydropyrone
R
O
CoA -S
O O
-CoASH
Flavonoids arise
from
condensation of
phenylpropanoid
+ triketide. The
first member
formed is
chalcone.
O
*
styryl pyrone
CO2-
1.
CoA -S
2. -CO2, -CoASH
R
R
OH
HO
to the
flavonoids
-CoASH
*
O
O
chalcone
S-CoA
*
O
*
OH
chalcone
synthase
(polyketide
cyclization)
33
5'
Oxidation
Level
8
7
4'
6'
1
3'
1'
2'
6
5
10
3
4
flavone
OH
O
dihydrochalcone
flavan-3-ol (catechin)
O
O
The principal
structural feature
of the flavonoids
is C6-C3-C6. The
flavonoids can be
subdivided by
oxidation level
and oxidation
position.
O
O
chalcone
O
flavanone
isoflavanone
+
O
OH
OH
flavan-3,4-diol
(leucoanthocyanidinin)
flavylium
34
Oxidation
level
O
O
O
O
isoflavone
O
flavone
aurone
+
O
OH
O
OH
anthocyanidin
flavanonol
(dihydroflavonol)
The principal
structural
feature of the
flavonoids is
C6-C3-C6.
Flavonoids can
be subdivided
by oxidation
level at the C
ring.
O
5
OH
O
flavonol
35
Luteolin
Kaempferol
Quercetin
2. Flavonols:
3. Flavanones:
Naringenin
8.0 Shikimates & phenylpropanoids (Dayrit)
36
5. Flavan-3-ols:
Epicatechin
Epigallocatechin
6. Anthocyanidins:
Cyanidin
8.0 Shikimates & phenylpropanoids (Dayrit)
37
OH
OH
OH
HO
OH
HO
naringenin
(flavanone)
OH
chalcone
4'
flavones
flavonols
7
5
OH
O apigenin
(flavone)
[O]
HO
OH
7
5
OH
OH
luteolin
4'
3'
HO
OCH3
7
5
OH
OH
OH
quercetinin
OH
4'
3'
HO
5
OH
[Me]
OH
anthocyanin
4'
3'
2
[Me]
OH
kaempferol
(flavonol)
HO
OH
OH
[O]
OH
4'
3'
OH
HO
HO
OCH3
OH
8.0 Shikimates
& phenylpropanoids (Dayrit)
chrysoeriol
OH
4'
4'
4'
HO
OH
OH
OH
Biosynthetic
studies in tissue
culture of parsley:
Chalcone is the
entry to the
flavonoids.
Flavonoids can be
further grouped
according to the
specific position
of oxidation at the
three rings. Note
that further
oxidation and
methylation occur
after flavonoid
ring formation.
isohamnetin
38
Occurrence of flavonoids
in various plants (Wink,
Biochemistry of Secondary
Metabolites 2010). There
are
some taxonomic patterns
but many questions remain.
OH
HO
OH
Glc-O
OH
OH
OH
UDP-glucose
UDP
OH
UDP-glucose
UDP
OH
OH
Glc -O
HO
OH
OH
O-Glc
O
O
2x CoASH
O
O
O
CO2
OH
2x malonyl CoA
OH
Numerous
flavonoids are
formed by further
modifications of
the basic
flavonoid rings.
For example,
glycosylation and
acetylation are
commonly
observed
modifications.
CO2
O
O
O
HO
OH
HO
bismalonyl isorhamnetin-3,7-bisglucoside
40
Isoflavonoids
Isoflavonoids are formed from flavonoids by 1,2-migration
of the B-ring from the 2- to the 3-position. Studies using
labeled phenylalanine are consistent with the proposed origin
of the isoflavonoids.
A. Labeling studies on isoflavonoids
OH
HO
*
# ^
HO2C
HO
NH2
*
# ^
*
# ^
O
OCH3
formononetin
41
HO
Isoflavonoids
OCH3
42
HO
Isoflavonoids
OCH3
43
Flavonoids
are converted
pterocarpan
enzymatically
to the
isomeric
isoflavonoids.
dehydropterocarpan
The
isoflavonoids
are found
mostly in the
coumestan
Leguminosae
family.
isoflavanone
O
O
isoflavone
O
3-arylcoumarin
O
rotenoid
dehydrorotenoid
O
O
44
B. Biosynthesis of rotenone
OPP
[O]
HO
1. [O]
2. ATP
H3C
isoflavone
HO
H2C
OCH3
OCH3
[H-]
OCH3
OCH3
NADPH
HO
HO
H
O
OPP
OCH3
H
OCH3
OCH3
OCH3
Rotenone is one
of the best
known of the
isoflavonoids.
Rotenone is
found in
mangrove
plants and is a
fish poison, but
is considered
safe for
mammals.
H
s
s
H
O
rotenone
OCH3
OCH3
45
R:
R5'
OH
+
O
HO
R3'
3'
5'
Name
Color
pelargonidin
red
OH
cyanidin
blue
OH
OH
delphinidin
deep purple
Glc
Glc
OH
cyanin
colorless
O-R3
O-R5
46
R5'
R5'
+
O
HO
OH
R3'
HO
+
OH
R3'
H
O R3
-
O-R3
O-R5
O-R5
non-planar conformation is colorless;
this form predominates if R3 is large
47
The yellow day lily has a near uniform hue in the visible
spectral range. However, in the UV range which is invisible
to most animals but visible to bees, the lily has a two-toned
pattern.
The UV pattern of many flowers is due to anthocyanins.
8.0 Shikimates & phenylpropanoids (Dayrit)
48
49
50
A.
OH
+
O
HO
Catechins are
OH
HO
OH
OH
OH
catechin
anthocyanidin
OH
B.
HO
OH
OH
HO
OH
OH
OH
OH
OH
(-)-epicatechin
(-)-catechin
OH
HO
OH
OH
HO
OH
OH
OH
OH
OH
(+)-epicatechin
(+)-catechin
flavan-3-ols which
are believed to be
biosynthetically
related to the
anthocyanidins and
may be produced
during the
extraction of
anthocyanidins.
The best known
catechins are ()catechin and ()epicatechin which
are major
constituents of tea.
51
R
HO
OH
OH
OH
OH
OH
HO
OH
n x
HO
O
OH
OH
OH
OH
OH
OH
(-)-epicatechin
HO
O
OH
OH
OH
52
53
MK biosynthetic pathways. (B) Alternative pathway. Green and blue bold lines indicate
two carbon units derived from C-5 and C-6 of glucose via different metabolic pathways.
Based on the annotation of the open reading frames of S. coelicolor A3(2), we presumed
that SCO4491 (prenylation), SCO4556 (methylation), SCO4490 (decarboxylation), and
SCO4492 (decarboxylation) would be involved in the late step of the MK biosynthetic
pathway.
56
57
Metabolic channeling of
biosynthesis: flavonoids
isoflavonoids phytoalexins
Biosynthesis of O-methylated
isoflavonoids in licorice, and
conversion to phytoalexins. (Jorgensen et
al., Curr Opinion in Plant Biol 2005, 8:280291)
58
Summary
Many of the aromatic compounds found in higher plants belong
to the shikimates. The shikimates have for a diverse range of
properties and functions in plants, ranging from structural
(lignins), flower and leaf coloration (anthocyanins and
flavonoids), scent (some aryl propanes), and plant defense
(flavonoids and isoflavonoids).
The shikimates can be grouped by C6-Cn classification:
C6: phenols and quinones
C6-C1: benzoic acids; benzaldehydes; benzyl alcohols
C6-C2: phenylethyl amines; styrenes; acetophenones
C6-C3: phenyl propanoids: cinnamaldehydes; aryl propanes;
lignins; lignans
C6-C3-C6: flavonoids; isoflavonoids; catechins
8.0 Shikimates & phenylpropanoids (Dayrit)
59
glucose
shikimic acid
Summary of
the shikimates
chorismic acid
prephenic acid
phenylalanine,
tyrosine
C6-C2: phenylethylamines
C6-C3: cinnamaldehydes
indole,
tryptophan
+3 x (C2)
C6: phenols,
quinones
C6-C3-C6: flavonoids
isoflavonoids
catechins
condensed
tannins