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8.

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

Shikimic acid is produced directly


from two D-glucose metabolites:
phosphoenol pyruvate (C3) and Derythrose-4-phosphate (C4) condense
to form the seven carbon heptulose.
Heptulose cyclizes to form 5dehydroquinate which loses water to
form shikimic acid.

H H

_
CO2

HO

_
CO2

OH

OH
OH

OH

5-dehydroquinate

3-dehydroshikimate

_
CO2

HO

OH
OH
shikimate

8.0 Shikimates & phenylpropanoids (Dayrit)

HOCH2
H

HO

O
H

Shikimic acid is converted to 5-enolpyruvylshikimate-3-phosphate,


which loses phosphate to yield chorismic acid and prephenic acid.
Chorismic acid is the branch point to a group of benzoic acid
metabolites and tryptophan. Prephenic acid leads to the phenyl
propanoids (C6-C3) and the flavonoids (C6-C3-C6), and the amino
acids phenylalanine and tyrosine.

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

quinones from shikimate + terpene:


OH

OH

OH

CO2H

OH
homogentisic acid

alkarinin
O
R

5. Polyketides (Dayrit)

ubiquinones: R = H, CH3 ; n = 4-13

Shikimates comprise a large group of aromatic natural products.


CO2H

CO2H

NH2
phenylalanine, R=H
tyrosine, R=OH

O
cinnamic acid

coumarin

CO2H
O

CO2H

phenylacetic acid

benzoic acid
O
flavonoid
O

O
lignan

8.0 Shikimates & phenylpropanoids (Dayrit)

Shikimic acid picks up three carbons from of phosphoenol pyruvate to


form chorismic acid and prephenic acid. Chorismic acid is converted to
prephenic acid via a concerted [3,3]-sigmatropic shift.
_
CO2

_
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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

C6, C6-C1 and C6-C2


The shikimate metabolites can be grouped according to the
number of carbons atoms in the side chain.
C6: phenols and quinones
C6-C1: benzoic acid derivatives, including tannins
C6-C2: phenyl ethyl compounds

8.0 Shikimates & phenylpropanoids (Dayrit)

Routes to benzoquinones:
A. Polyketides

Hydroxybenzoic acids

B. Shikimatic acid

Benzoquinones

Homogentisic acid

C6 Metabolites

Benzoquinones

Chroismic acid
p-Hydroxybenzoic acid

Phenylalanine

Benzoquinones

From p-Hydroxybenzoic acid:


CO 2H

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

12

Ellagitannins are hydrolyzable


tannins which are formed from
the condensation of a sugar
core and ellagic acid units.
Ellagitannis from banaba,
Lagerstroemia speciosa, were
shown to increase uptake of
adipocytes in rats, and could be
responsible for lowering the
blood glucose level. (Hayashi, et
al., Ellagitannins from Lagerstroemia
speciosa as Activators of Glucose
Transport in Fat Cells, Planta Med.,
2002, 68, 173-5.)

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

Lagerstroemin: R1=OH; R2=H


Flosin B: R1=H; R2=OH

8.0 Shikimates & phenylpropanoids (Dayrit)

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

pro tocatec huic acid

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

Cyanogenic glycosides: C6-C2


A cyanogenic glycoside has an
aglycone with a cyanide group and
an attached sugar. Cyanogenic
glycosides release the poisonous
hydrogen cyanide enzymatically.
Cyanogenic glycosides are found in
cassava, and the fruits and wilting
leaves of the rose family (including
cherries, apples, plums, almonds,
peaches, apricots, and raspberries).
Sorghum (Sorghum bicolor)
expresses cyanogenic glycosides in
its roots and thus is resistant to pests
such as rootworms.
The dotted arrow is a metabolic link
to aromatic glucosinolates.

(Jorgensen et al., Curr Opinion in Plant Biol 2005, 8:280291)

8.0 Shikimates & phenylpropanoids (Dayrit)

15

Glucosinolates

The glucosinolates are found in

OH

HO

HO

OSO3
S

HO

HO

(R group
variable)

Brassicales and contain sulfur,


nitrogen and a group derived from
glucose.
About 120 glucosinolates are known
in plants, where the R group is alkyl,
derived from methionine, alanine,
leucine, or valin; aromatic, derived
from phenylalanine and tyrosine; or
indolic derived from tryptophan.

Glucosinolates act as natural pesticides and defense against

herbivores. These substances are also responsible for the bitter


or sharp taste of many common foods such as mustard, radish,
horseradish, cress, cabbage, Brussels sprouts, cauliflower,
broccoli, and turnip.
8.0 Shikimates & phenylpropanoids (Dayrit)

16

Phenyl propanoids: C6-C3


The phenylpropanoid metabolism is unique to plants.
Many intermediates and end products of the phenylpropanoid
pathway play important roles in plants as phytoalexins,
antioxidants, antiherbivory compounds, UV protectants,
pigments, and aroma compounds. Phenylpropanoids polymerize
to form lignins, which are essential components of the cell wall
stability.
Phenylpropanoid biosynthesis is one of the best-studied
pathways in plants. The enzymes of the phenylpropanoid pathway
are organized in multi-enzyme complexes and there is evidence
for the coordinated expression of genes and enzymes. Genes
encoding enzymes of this pathway are developmentally and
tissue-specifically regulated and may be induced by
environmental stresses such as nutrient deficiency, exposure to
cold, UV light, and8.0pathogen
attack.
Shikimates & phenylpropanoids
(Dayrit)
17

Phenylalanine and tyrosine are deaminated by phenylalanine ammonia lyase


(PAL) or tyrosine ammonia lyase (TAL) to cinnamic acids. The cinnamic acids
(C6-C3) are the precursors to the phenyl propanoids, coumarins, styrenes,
benzoic acids, phenols, and flavonoids. A multi-enzyme complex enables
coordinated action of PAL and cinnamate-4-hydroxylase (C4H) which control
the flux of intermediates in phenylpropanoid biosynthesis.
NH2

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

Ar - C1: benzoic acids


benzyl aldehydes
benzyl alcohols

8.0 Shikimates & phenylpropanoids (Dayrit)

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

3-caffeoylquinic acid (chlorogenic acid).


First isolated in 1846 from coffee, it has since
been found to be a common plant compound.
It functions as an allelopathic substance in
sunflower.

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

8.0 Shikimates & phenylpropanoids (Dayrit)

19

Examples of cinnamic acids and the C6-C3 derivatives. Rosmarinic acid is


a dimer of two different C6-C3 units.
CO2H

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

Coumarins are aromatic lactone C6-C3 derivatives. They are widely


distributed in plants, particularly the Umbelliferae and Rutacea families.
CO2H
[O]
RO

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

Examples of coumarin compounds.


O

O-H

CO2H
H

OH

HO

O-H

Some microorganisms (such as Aspergillus fumigatus)


transform coumarin from plants (such as grass)
to produce dicoumarol. Dicoumarol is a powerful
blood anticoagulant and can cause fatal hemmorages
in cattle that eat the hay.

CH2

dicoumarol

HO

OH
O

Furobinordenatin is a coumarin from Citrus plants.

O
O

HH

8.0 Shikimates & phenylpropanoids (Dayrit)

21

Lignans are phenylpropanoid dimers. They can be rationalized by formation by


coupling of resonance-stabilized by radicals with the assistance of dirigent proteins.
OH

oxidase with guiding (dirigent) protein

oxidase only
OCH3
OH

OH

O
CH3O

HO

CH 3O

OH

OH

OCH3

OCH3

OH
H

Proposed coupling intermediate

(+/-)-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);

Plant Physiol., 123, 453 (2000); Chem. Biol., 6, 143 (1999))

22

The diversity of lignan structures can be rationalized by bond formation via


different radical sites.
CH 2OH

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

8.0 Shikimates & phenylpropanoids (Dayrit)

OCH3
OH
(+)isolariciresinol

23

The diversity of lignan structures can be rationalized by bond formation via


different radical sites.
H

.
CH3O

.
CH3O

O
O

OCH3

CH3O

O
OCH3

H+

O
OCH3

HO
licarin A

(from eugenol)

8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

26

Intermediates and enzymes of the lignin pathway.

4CL, 4-(hydroxy)cinnamoyl CoA ligase


C3H, p-coumarate 3-hydroxylase
C4H, cinnamate 4-hydroxylase
CAD, cinnamyl alcohol dehydrogenase
CCoAOMT, caffeoyl CoA O-methyltransferase
CCR, cinnamoyl CoA reductase
COMT, caffeic acid/5-hydroxyferulic acid O-methyltransferase
CQT, hydroxycinnamoyl CoA:quinate hydroxycinnamoyltransferase
CST, hydroxycinnamoyl CoA:shikimate hydroxycinnamoyltransferase
F5H, ferulate 5-hydroxylase
PAL, phenylalanine ammonia-lyase
pCCoA3H, p-coumaryl CoA 3-hydroxylase
SAD, sinapyl alcohol dehydrogenase.

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

Flavonoids have attracted wide interest due to the following:


a. Distribution of flavonoids. The flavonoids are one of the
most numerous and widespread groups of secondary
metabolites. They are present in most parts of the plant, often
in various methylated and glycosylated forms.
b. Functions of flavonoids in plants. The colored pigments in
flowers of angiosperms (blues, purples, reds, and yellows)
yellows
are produced by the flavonoid group known as the
anthocyanidins. In leaves, the chlorophylls mask these
colors. The highly oxygenated flavonoids are UV-active
and are thought to protect the plant against UV radiation.
Various flavonoids are believed to play specific roles in the
physiology of plants. A number of flavonoids, for example,
are known to be antifungal, antibacterial, and allelopathic.
8.0 Shikimates & phenylpropanoids (Dayrit)

29

OH

Flavonoids

HO

OH

OH

luteolin

c. Biological activity of flavonoids. A number of flavonoids


have been shown to protect cells against oxidative attack.
This is thought to explain its anti-inflammatory and cancerprotective properties. In addition, flavonoids have been
shown to strengthen blood capillaries, and have found use
in protection against weak arteries, strokes and
hemorrhoids.
Flavonoids are formed from the condensation of a
phenylpropanoid + triketide. The flavonoids can be
subdivided according into oxidation level and position at
three rings.
the
Extensive
biosynthetic studies have been carried out using
tissue culture of parsley. Chalcone is the entry point to the
flavonoids. Further oxidation and methylation occur after
flavonoid ring formation.
8.0 Shikimates & phenylpropanoids (Dayrit)

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

The first member of


the flavonoid group is
chalcone.

*
O

(C6-C3) + C6.
R

*
O

CoA -S

2. -CO2, -CoASH

dihydropyrone

R
O

(C6-C3) + C4;

OH

(C6-C3) + C2;

-CoASH

Flavonoids arise from


condensation of a
phenylpropanoid, (C6C3), with three acetyl
CoA units (3 x C2).
The addition of acetyl
CoA probably occurs
stepwise yielding
intermediates with the
following structures:

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

35

Six flavonoid subgroups


1. Flavones:
Apigenin

Luteolin

Kaempferol

Quercetin

2. Flavonols:

3. Flavanones:
Naringenin
8.0 Shikimates & phenylpropanoids (Dayrit)

36

Six flavonoid subgroups


4. Isoflavones:

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

OCH3

formononetin

41

HO

Isoflavonoids

OCH3

The isoflavonoids, which are produced in legumes, are


essential for plant-microbe interactions, such as chemoattractants and signal molecules for symbiotic Rhizobium
bacteria. The nodD gene-encoded proteins of Rhizobia
have been shown to physically bind to flavonoids and
isoflavonoids, and this ligand association initiates
transcription of the nod operon leading to root nodule
formation.
Different isoflavonoids are also either precursors to, or are
themselves the major phytoalexins in legumes, which play
key roles in non-specific plant defense against bacterial
and fungal pathogens. The activation of isoflavonoid
synthesis during the disease resistance response is
important for providing these many defense compounds.
8.0 Shikimates & phenylpropanoids (Dayrit)

42

HO

Isoflavonoids

OCH3

Isoflavonoid synthesis is a branch of the general


phenylpropanoid pathway that exists in all higher plants.
An enzyme found almost exclusively in the legumes,
isoflavone synthase (IFS), converts the phenylpropanoid
pathway intermediates tetrahydroxy-chalcone (naringenin)
and trihydroxy-chalcone (isoliquiritigenin) into the
isoflavones genistein and daidzein, respectively.
Isoflavones are produced at significant levels only in
tissues where the phenylpropanoid pathway activity is
elevated, such as in floral tissues, in UV-treated tissues,
and in tissues where expression of a heterologous
transcription factor was used to activate genes of the
phenylpropanoid pathway.
8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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.

another closely related compound: milletone

H
s
s

H
O

rotenone

OCH3
OCH3

8.0 Shikimates & phenylpropanoids (Dayrit)O

45

Anthocyanins are the sugar- derivatives of the anthocyanidins


(the aglycone). Anthocyanins are water-soluble pigments in
flowers, leaves and fruits. They also impart many of the colors of
fruit juices and wines.
A. List of the best-known anthocyanins and anthocyanidins;
A. Substituents and Color

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

8.0 Shikimates & phenylpropanoids (Dayrit)

46

B. Anthocyanin color is produced in compounds which can


achieve a planar conformation which gives maximum
delocalization.
B. Structure and Color

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

extended resonance form is colored;


this conformation is possible if R3 is small

8.0 Shikimates & phenylpropanoids (Dayrit)

47

Anthocyanins and flower color


(The Big Bloom: Birth of
Flowering Plants, National
Geographic, July 2002)

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

Leaf color is due to several


Anthocyanins and autumn leaves compounds:

Why do leaves turn yellow, orange, Chlorophyll, which is green, gives


leaves its predominant color. It is
red, purple and magenta during
responsible for photosynthesis. As the
autumn?

temperature drops and the days


shorten, it is destroyed enzymatically,
revealing the presence of the other leaf
pigments.
Carotenoids are responsible for the
yellow, orange and gold hues.
Different species of trees contain
various types of carotenoids.
Anthocyanins give rise to the reds,
purple and magenta hues. In contrast
to the carotenoids, which are always
present, anthocyanins are actively
formed in the leaves during autumn.

8.0 Shikimates & phenylpropanoids (Dayrit)

49

Anthocyanins and autumn leaves


Why do leaves turn yellow, orange, What is the special role of
anthocyanins?
red, purple and magenta during
autumn?
It is hypothesized that
anthocyanins protect the leaf from
free radicals which are produced
by the destruction of the
photosynthetic system.
They provide some
environmental protection from
UV radiation and nutrient
deficiency.
The net effect is that
anthocyanins slow down leaf
death and abscission.
8.0 Shikimates & phenylpropanoids (Dayrit)

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

8.0 Shikimates & phenylpropanoids (Dayrit)

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

Condensed tannins are polymeric compounds made up of


catechin monomers.
OH

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

8.0 Shikimates & phenylpropanoids (Dayrit)

52

8.0 Shikimates & phenylpropanoids (Dayrit)

53

MK biosynthetic pathways. (A) Classical pathway. Chorismate is converted into


isochorismate by MenF (isochorismate synthase) and then into 2-succinyl-6-hydroxy-2,4cyclohexadiene-1-carboxylate by MenD. This compound is dehydrated by MenC to osuccinylbenzoate, followed by the attachment of coenzyme A to yield o-succinylbenzoylCoA by MenE. o-Succinylbenzoyl-CoA is then converted into 1,4-dihydroxy-2-naphthoate
by MenB. In the last two steps of the pathway, MK is converted by MenA and MenG,
which catalyze prenylation and methylation, respectively. Red and black bold lines show
carbons originated from erythrose-4-phosphate and phosphoenolpyruvate, respectively.

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.

Metabolons of Phenylpropanoid Genes


A metabolon is a temporary functional
complex of several sequential enzymes
held together by noncovalent
interactions in a given metabolic
pathway.
Organization of the branched pathways
of phenylpropanoid metabolism within
separate individual metabolons.

Jorgensen et al., Curr Opinion in Plant Biol 2005, 8:280291


4CL, 4-(hydroxy)cinnamoyl CoA ligase
C3H, p-coumarate 3-hydroxylase
C4H, cinnamate 4-hydroxylase
CAD, cinnamyl alcohol dehydrogenase
CCoAOMT, caffeoyl CoA O-methyltransferase
CCR, cinnamoyl CoA reductase
8.0

Enzymes that participate in multiple


branches are shown in red and blue, whereas
enzymes that are thought to be unique to
specific pathways are shown in other colors.
Enzymes that are known to be present as
multiple isoforms are marked with an asterix.

COMT, caffeic acid/5-hydroxyferulic acid O-methyltransferase


CQT, hydroxycinnamoyl CoA:quinate hydroxycinnamoyltransferase
CST, hydroxycinnamoyl CoA:shikimate hydroxycinnamoyltransferase
F5H, ferulate 5-hydroxylase
PAL, phenylalanine ammonia-lyase
Shikimates
pCCoA3H&
, p-coumaryl
phenylpropanoids
CoA 3-hydroxylase
(Dayrit)
SAD, sinapyl alcohol dehydrogenase.

56

Metabolons and the Biosynthesis of Natural Products


Metabolon formation and metabolic channeling in plant secondary
metabolism enable plants to effectively synthesize specific natural
products and to avoid metabolic interference.
Channeling can involve different cell types, take advantage of
compartmentalization within the same cell or proceed directly within a
metabolon. New experimental approaches document the importance of
channeling in the synthesis of isoprenoids, alkaloids,
phenylpropanoids, flavonoids and cyanogenic glucosides.
Metabolon formation and metabolic channeling in natural-product
synthesis facilitate attempts to genetically engineer new pathways into
plants to improve their content of valuable natural products. They also
offer the opportunity to introduce new traits by genetic engineering to
produce plant cultivars that adhere to the principle of substantial
equivalence. (Jorgensen et al., Curr Opinion in Plant Biol 2005, 8:280291)
8.0 Shikimates & phenylpropanoids (Dayrit)

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)

IFS: 2-hydroxyisoflavanone synthase


HI4OMT: 2,7,4-trihydroxyisoflavone 4-Omethyltransferase
D7OMT: daidzein 7-O-methyltransferase
HID: 2-hydroxyisoflavanone dehydratase
8.0 Shikimates & phenylpropanoids (Dayrit)

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

C6-C1: p-hydroxybenzoic acid

chorismic acid

C6-C1: o-hydroxybenzoic acid


(salicylic acid)
C6-C1: p-aminobenzoic acid

prephenic acid

phenylalanine,
tyrosine

C6-C2: phenylethylamines

C6-C3: cinnamic acids

C6-C3: cinnamaldehydes

C6-C3: cinnamic alcohols


C6-C3: coumarins
C6-C3: arylpropanes

C6-C1: o-aminobenzoic acid


(anthranilic acid)

indole,
tryptophan
+3 x (C2)

C6: phenols,
quinones

C6-C1: benzoic acids,


C6-C2: styrenes,
benzaldehydes,
benzyl alcohols
acetophenones

C6-C3-C6: flavonoids

isoflavonoids

catechins

condensed
tannins

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