KOE 322: Natural Product

Chemistry
Phenolic compounds
Prepared by Dr. Thiruventhan Karunakaran
Centre for Drug Research USM
Ext 3287
thiruventhan@usm.my
 Phenolic compounds (Plant source- phytochemistry)

• Phenolic compounds compromise the largest group of phytochemicals and are distributed widely in the plant
kingdom.

• Hydrocarbons in which one or more hydrogen atoms are replaced by hydroxyl group (-OH) are named as
alcohols and phenols.
• In other words, the hydroxyl derivatives of hydrocarbons are named as alcohols and phenols. Out of alcohols
and phenols, alcohols are hydroxyl derivatives of aliphatic hydrocarbons, whereas phenols are hydroxyl
derivatives of aromatic hydrocarbons.
• In organic chemistry, phenols are a class of chemical compounds consisting of a hydroxyl group (-OH) bonded
directly to an aromatic hydrocarbon group.
• Phenolic compounds are widely distributed in plant tissues, particularly contributing colour, flavour, and
astringency to fruits.
• The concentration of phenolic compounds may vary from 0.5 to 5.0 g per 100 g dry weight of plant tissues.
• Phenolic compounds are often considered secondary metabolites of plant metabolism that contribute little to the
physiological or ecological functions of the plant.
• Most phenolic compounds are believed to be by-products of the metabolism of the aromatic amino acid
phenylalanine.
• A major group of water-soluble phenolic compounds, the anthocyanins, contribute colours to fruits.
 Phenolic compounds (Plant source- phytochemistry)

• Flavonoids, phenolic acids and polyphenols are the three main subgroups in the phenolic group.
• The simplest class of the natural phenolic compounds is the phenol (C 6H5OH).
• Phenolic compounds play a vital role in plants’ defense mechanism.
• Moreover, phenolic compounds exhibit many properties especially medicinal properties to humans as well
as antioxidant properties that are pivotal and act as protecting agents against free radical mediated
disease processes (Dai et al., 2010; Balasudram et al.,2006; Walton et al., 2003).
• Benzopyrone groups such as coumarin, flavonoids and xanthones are the principal members in the
phenolic type compounds.
• This benzopyrone group of compounds possess many pharmacological and medicinal properties.
• The precursor for the formation of many phenolic compounds is shikimic acid.
• Most phenolic compounds were synthesized in plant metabolism through the shikimic acid pathway.
• Draw the respective structure of the compounds.
 The Most Important Classes of Phenolic Compounds in Plants

number of C-atoms basic skeleton class

6 C6 simple phenols, benzoquinones

7 C6 - C1 phenolic acids

8 C6 - C2 acetophenone, phenylacetic acid

hydroxycinnamic acid, polypropene,

9 C6 - C3
coumarin, isocoumarin

10 C6 - C4 naphtoquinone

13 C6 - C1 - C6 xanthone

14 C6 - C2 - C6 stilbene, anthrachinone

15 C6 - C3 - C6 flavonoids, isoflavonoids

18 (C6 - C3)2 lignans, neolignans

30 (C6 - C3 - C6)2 biflavonoids

(C6 - C3)n lignins

n (C6)n catecholmelanine
(C6 - C3 - C6)n (condensed tannins)
Phenyl-Propanoid Natural Products (C6-C3)

• The biosynthesis of the aromatic amino acids occurs through the shikimic acid pathway, which is
found in plants and microorganisms (but not in animals).
• Humans require these amino acids in our diet, since we are unable to produce them.

• The aromatic amino acids also serve as starting materials for the biosynthesis of many interesting
natural products.
Here we will focus on the so-called phenyl-propanoide (C6-C3) natural products, e.g.:
Enzyme : Biocatalyst
• Enzymes: are group of molecules that serve as a catalyst with a high degree of specificity for a
certain substrate or class of substrates. It can only act on one substrate or on a family of structurally
similar substrates.
• Major Enzymes :
1. Synthase: Joints two molecules together w/o hydrolyzing a pyrophosphate bond.
2. Dehydratase: Removes water to create a double bond
3. Dehydrogenase: Removes hydrogen atom from its substrate
4. Kinase: Transfer a phosphate group from a high-energy phosphate compound such as
ATP to its substrate.
• More…….
 Shikimic Acid
• Where this came from?
• Found the genus Lllicium
Shikimic Acid / Shikimate Pathway

• Shikimic acid is a precursor for:

1. Aromatic amino acids phenylalanine and tyrosine.
2. Indole and indole derivatives and eg: tryptophan – Alkaloids.
3. Phenylpropanoids, flavonoids, tannins, and lignins.
• A very large number of compounds exhibit a characteristic C6-aromatic-C3-side chain structure. E.g.
aromatic amino acids, cinnamic acids, coumarins, flavonoids, lignin constituents, etc.
• These come from a common origin.
• It was found that erythrose-4-phosphate starts the biosynthetic pathway leading to shikimic acid.
• The biosynthesis of these compounds was elucidated by mutant studies of E.coli by Davis and
Sprinson.
• Shikimic acid was isolated as early as 1885 by Eykman from the Japanese plant, Illicium
anisatum long before we were aware of its biosynthetic significance.
• The biosynthesis pathway begins with D-erythrosephosphate and phosphoenolpyruvate (PEP)
combining via an aldol condensation.
• 3-deoxy-D-arabinoheptulosonate 7-phosphate (DAHP) synthase is the first enzyme in a series of
metabolic reactions known as the shikimate pathway. Since it is the first enzyme in the shikimate
pathway, it controls the amount of carbon entering the pathway.
• Both these compounds were initially derived from D-glucose.
• The aldol condensation is aided by an enzyme which adds on to the phosphoenolpyruvate molecule to
form 3-deoxy-D-arabinoheptulosonate-7-phosphate (DAHP)
• Ring closure of the heptulose derivative (3,7-dideoxy-D-arabino-2,6-diulosonic acid) gives 3-
dehydroquinic acid.
• 3-dehydroquinate synthase is the second enzyme of the shikimate pathway. It catalyzes the elimination of
phosphate from DAHP to generate 3-dehydroquinic acid (DHQ).
• Removal of 1 H2O molecule from 3-dehydroquinic acid yields 3-dehydroshikimic acid; this acid is reduced
to shikimic acid.
• At the pH of living organism, this acid exists in its anionic form, the shikimate ion.
Shikimic Acid Formation-
Shikimate Pathway:
Shikimic Acid Formation -
Shikimate Pathway:
Shikimic Acid Formation-
Shikimate Pathway: H phosphoenol pyruvate
COOH
O O C
H O COOH
HO P O C CH2
CH2 B: H
O HO OH
H
O
H H+ H
HO O HO P O CH2 HO
H
O
PO CH2 HO H

erythrose-4-phosphate

H+ H+ COOH
HO
COOH COOH COOH O C
CH2
NADPH H
H2C H
OH
HO OH O OH O OH
OH HO H
OH OH H O

shikimic acid
3-deoxy-D-arabinoheptulosonate 7-phosphate (DAHP) synthase is the first enzyme in a
series of metabolic reactions known as the shikimate pathway. Since it is the first enzyme in
the shikimate pathway, it controls the amount of carbon entering the pathway.
3-dehydroquinate synthase is the second enzyme of the shikimate pathway. It catalyzes
the elimination of phosphate from DAHP to generate 3-dehydroquinic acid (DHQ).
• Phosphorylation of the shikimate anion with adenosine triphosphate gives shikimate-3-phosphate
which then reacts with another molecule of phosphoenolpyruvate (PEP) to yield 5-
enolpyruvylshikimate-3-phosphate.
 O O- O O-
O O-
-H+
enzyme
surface H+
P O OH P O O- O CH2
OH OH P
shikimate-3-phosphate PEP

O O- O O-
O O- O O-

enzyme

O H
P O O CH2 P O
O H
OH OH H Enz
P
5-enolpyruvylshikimate-3-phosphate
Shikimate-5-Dehydrogenase (SDH) The fourth step of the shikimate pathway is the
reduction of DHS to shikimate. the reaction is catalyzed by an NADP-dependent shikimate
dehydrogenase (SHD)
Shikimate Kinase (SK)- In the fifth step of the shikimate pathway, shikimate
kinase catalyzes the specific phosphorylation of the 3-hydroxyl group of D-
shikimate to yield shikimate 3-phosphate using ATP as a co-subtrate
• 5-enolpyruvylshikimate-3-phosphate converts to chorismate by elimination (1,4 with respect to
Hydrogen and Phosphate).
• Enzyme assistance is once more involved in this conversion.
• Chorismate Synthase (CS). The seventh and final step in the main trunk of the shikimate pathway is
the trans-1,4 elimination of phosphate form EPSP to yield chorismate.
• In this reaction, the second of the three double bonds of the benzene ring is introduced. The reaction is
catalyzed by chorismate synthase and requires reduced flavin for activity even though the overall
reaction is redox neutral.
• Most phenolic compounds belong to the flavonoids.

• Lignin,the primary substance of wood, is the most common

member of this group.

• The starting product of the biosynthesis of most phenolic compounds is

shikimate.

• Among the phenylpropanol derivatives of lower molecular weight are a number

of scents like the coumarins, cinnamic acid, sinapinic acid, the coniferyl alcohols
and others.

• These substances and their derivatives are at the same time intermediates
of the biosynthesis of lignin.
Precursors for plant phenolic compounds
The phenylpropanoids: products of the shikimic acid pathway
 The phenylpropanoids: products of the shikimic acid pathway (phe and tyr)

The shikimate
pathway
Cinnamic acid and Benzoic acid
• The widely distributed cinnamic acids arise from phenylalanine by enzymatic elimination of
ammonia followed by aromatic hydroxylation and methylation.

• Shortening of the side chain of cinnamic acids by β- oxidation is one of the leading routes to benzoic
acids.

• Hydroxylation is more effective at the C6C3 level than at C6C1 level in plants.

• Hence, to reach at vanillic acid, the route goes from coumaric acid, caffeic acid, ferulic acid, p-OH-
benzoic acid and protocatechic acid.
Phenolics can be classified into 2 groups:

1. The FLAVONOIDS

2. The NON-FLAVONOIDS
Flavanoids