Cofactor and Coenzyme

§ Cofactor is organic molecule or metal ion which the

enzymes require in order to catalyse a reaction

§ Cofactors can be categorized into two groups- organic

cofactors and inorganic cofactors.

§ Coenzymes are organic cofactors which are again

divided into two groups- co-substrates and prosthetic
groups.
Introduction § Coenzyme which bound loosely to an enzyme are
termed as co-substrate and coenzyme which bound
tightly to an enzyme are termed as prosthetic groups.

§ Essential ions are inorganic cofactors which is again

classified as- activator ions (loosely bound) and
metalloenzymes (tightly bound).
 reagents. Most coenzymes are linked to enzymes by noncovalent forces. Coen
separated from the enzyme by dialysis. Essential ions are inorganic cofactors
classified as- activator ions (loosely bound) and metalloenzymes (tightly bound
§ An enzyme without cofactor is called apoenzyme and the enzyme-cofactor complex is called
An enzyme
holoenzyme. Apoenzyme without cofactor
is enzymatically is called apoenzyme and the enzyme-cofactor com
inactive protein.
holoenzyme. Apoenzyme is enzymatically inactive protein.

Organic Cofactors= Coenzymes

Inorganic Cofactors= Essential ions

Apoenzyme= Enzyme - Cofactor

Holoenzyme= Enzyme + Cofactor

A coenzyme creates an active site for catalytic activity:

Apoenzyme= Enzyme - Cofactor

Holoenzyme= Enzyme + Cofactor

A coenzyme creates an active site for catalytic activity:

 Types of cofactors

*ATP
Ca++ Fe-S center * FMN/FAD
K+ * SAM
zinc
UDP-sugar * TPP
Mg++ copper
* NAD+/NADP+ * PLP
Mn++ cobalt
* tetrahydrofolate * Biotin
See:
* CoA * adenosyl/methyl-
*pg. 126/132 for info on metal ions cobalamin
in catalysis * ubiquinone
*Table 15.2 as summary table for protein coenzymes
* Lipoic acid/
coenzymes lipoamide
§ There are two types of coenzymes: Metabolite Coenzyme and Vitamin derived Coenzyme

§ Metabolite coenzymes- These are generally derived from common metabolites usually from
nucleotides. The most commonly used metabolite coenzyme is adenosine triphosphate (ATP).

§ Vitamin derived coenzyme- These are synthesized by microorganisms and plants, generally
obtained from nutrients. Most vitamins must be enzymatically transformed to the coenzyme.
Some vitamins directly act as coenzymes but some vitamins help the body to produce
coenzymes.
 Metabolite coenzymes- These are generally derived from common metabolites usually from
nucleotides. The most commonly used metabolite coenzyme is adenosine triphosphate (ATP).

Vitamin derived coenzyme- These are synthesized by microorganisms and plants, generally
obtained from nutrients. Most vitamins must be enzymatically transformed to the coenzyme.
§ Some Some vitamins directly act as coenzymes but some vitamins help the body to produce
vitamins with their coenzymes are listed below:
coenzymes.

Some vitamins with their coenzymes are listed below:

Vitamins Coenzymes

Niacin NAD(P)+NAD(P)H

Thiamin (B1) Thiamin-pyrophosphate

Riboflavin (B2) FMN & FAD

Pantothenate (B5) Coenzyme A

Pyridoxal (B6) Pyridoxal phosphate

Cobalamin (B12) Adenosyl and methylcobalamin

Ascorbic acid(C) Not a coenzyme

Vitamin A Retinal

Vitamin K Vitamin K

Biotin/Vitamin H Biotin

Folate Tetrahydrofolate

Here we have discussed about some coenzymes in the following:

NAD+ & NADP+:

Both functions as an acceptor of hydrogen atoms and electrons in presence of

 *
2 electrons
1 or 2 electrons
1 or 2 electrons

2 carbon groups
containing carbonyl 
ATP‐dependent 
carboxylation

yl-

*ATP may also donate pyrophosphoryl and adenosyl groups

 Biotin/Vitamin H Biotin

Folate Tetrahydrofolate

NAD+ & NADP+Here we have discussed about some coenzymes in the following:
NAD+ & NADP+:
§ Both functions as an acceptor of hydrogen atoms and electrons in presence of
Both functions as an acceptor of hydrogen atoms and electrons in presence of
dehydrogenases and converted
dehydrogenases and converted into
into thethe reduced
reduced form. form.
O
O H H

NH2
NH2
+H-

N+ N
-H-
R R

NAD+ NADH
Oxidised Reduced

The oxidized form of coenzymes accepts pairs of electrons whereas the reduced form of
§ The oxidized coenzymes
form ofdonate pairs of electrons. They act as cosubstrates for dehydrogenase and also
coenzymes accepts pairs of electrons whereas the reduced form of
functions as hydride ion transfer. Niacin is required for the synthesis of these two coenzyme
coenzymes donate
moleculespairs
NAD+ &ofNADP+.
electrons.
The structures of NAD+ and NADP+ are given below:
§ They act as cosubstrates for dehydrogenase and also functions as hydride ion transfer.
NH2
O
§ Niacin is required for the synthesis of these two coenzyme molecules NAD+ & NADP+.
N
N
Adenine

O
N
CH2 OH
O
Phosphate D-ribose
HO P O

H H
N
OH OR
O
CONH2

Nicotinamide
Niacin
molecules NAD+ & NADP+. N+
-H-
N

R R

The structures of NAD+ and NADP+ are given below: NAD+

Oxidised
NADH
Reduced

NH2 The oxidized form of coenzymes accepts pairs of electrons whereas the reduced form of
O
coenzymes donate pairs of electrons. They act as cosubstrates for dehydrogenase and also
N
functions as hydride ion transfer. Niacin is required for the synthesis of these two coenzyme
N molecules NAD+ Adenine
& NADP+.
N
The structures of NAD+ and NADP+ are given below:
O
N
CH2 NH2
OH
O O
N

Phosphate D-ribose
N Adenine
HO P O
N
H H N
OH
O CH2
O N
OH OR
Phosphate HO P O
D-ribose

H H
O N
CONH2

O
Nicotinamide
OH OR

CONH2
Niacin
Phosphate HO P O
Phosphate HO P O
Nicotinamide (Nicotinic
Niacinacid)
(Nicotinic acid)
N+ N+
O CH2
O CH2
O
O
D-ribose
H

D-ribose Thiamin-pyrophosphate:
HO OH
H
NAD+ : R=H
Thiamin-pyrophosphate:
NADP+ : R=PO3H2
It is a coenzyme which participates in
HO OH

NADP+ : R=PO3H2
NAD+ : R=H
It is a coenzyme which participates in
 NOTE: Electron carrier in 
many oxidation reduction 
reactions; used for 
NAD+ reactions in pathways that 
lead to generation of ATP

Nicotinamide adenine dinucleotide

_____________________

Fig. 15‐12a/ 
15.13
 Flavine ade
Nicotinamide adenine 
dinucleotide phosphate

__________________

NOTE: Electron donor in 
most reductive 
biosynthesis reactions

Fig. 15.15/15.16
Fig. 15.13/15.14
Thiamin-pyrophosphate (transfer aldehyde)

§ It is a coenzyme which participates in oxidative decarboxylation reactions a-keto carboxylic

acids and transketolase enzyme reaction.
oxidative decarboxylation reactions and transketolase enzyme reaction. It also catalyzes
several biological reactions. It is derived from Thiamin (vitamin B1).
§ It also catalyses several biological reactions and derived from Thiamin (vitamin B1).
The structure of thiamin pyrophosphate is given below:

Dissociable proton

NH2 H
H2
C
N N+
S

O O
H3C N
CH2
CH2
O P O P O-

O- O-

Thiamin pyrophosphate (TPP)

FMN & FAD:

Riboflavin (vitamin B2) is required for the synthesis of flavin nucleotides (FMN & FAD).
FAD/FMN

§ Riboflavin (vitamin B2) is required for the synthesis of flavin nucleotides (FMN & FAD).
§ Generally FAD function as prosthetic groups as they are noncovalently bonded to the enzyme
and therefore cannot dissociate.

§ They act as temporary storage of electrons within the proteins and also function in the
oxidation/reduction reaction.
 They can transfer electrons one or two at a time.
§ They can transfer electrons one or two at a time.

Pyridoxal-phosphate:

It is a coenzyme which is derived from vitamin B6. It functions in isomerisation,

 Figure 02: Structures of FAD and FADH

What are the similarities between NADH and FADH2?

NADH and FADH2 are coenzymes
Both act as electron carriers.
Both are nonprotein organic molecules.
Both are derived from vitamins.
Both are water soluble.
Both can exist in the reduced form or oxidized form.
Both participate in oxidation and reduction reactions and help in the transfer of
electrons from one substrate to the other.
Both coenzymes can be synthesized in the body.
Both molecules take part in metabolic pathways which include carbohydrate,
fatty acid, amino acid and nucleotide metabolism.

What is the difference between NADH and FADH2?

NADH vs FADH2
NADH is a coenzyme derived from vitamin FADH2 is a coenzyme derived from
B3 or niacin. Vitamin B2 or riboflavin.

ATP Produced

NADH gives 3 ATP. NADH gives 2 ATP.

Commercial Applications

NADH is used as a supplement under This has no commercial applications.

energy deprived conditions.

Summary – NADH vs FADH2

Pyridoxal-phosphate

§ It is a coenzyme which is derived from vitamin B6.

§ It functions in isomerisation, decarboxylation, racemization, transamination, side chain
elimination reaction involving amino acids.
Biotin

§ Biotin is a coenzyme which is derived from vitamin H.

Biotin:
§ It functions in the metabolism of fats and amino acids & also acts as a carrier of activated CO2.
Biotin is a coenzyme which is derived from vitamin H. It functions in the metabolism of fats
and amino acids & also acts as a carrier of activated CO2.
O

HN NH

H H

COOH
S

Biotin

Tetrahydrofolate:

Tetrahydrofolate is derived from vitamin folic acid. It is required in the biosynthesis of

purines and pyrimidines.
O COO-

N COO-

H
OH H NH
Pantothenic Acid (Vitamin B5)
§ Pantothenic acid is part of coenzyme A
§ Coenzyme A is involved in energy production, conversion of lipids and amino acids to glucose
and synthesis of cholesterol and steroid hormones
§ Coenzyme A is one of the key coenzymes in the bodies biochemistry and is vital to the citric
acid cycle for production of ATP.

§ Responsible for acyl transfer

 HN NH

H H

COOH
S

Tetrahydrofolate (transfer one carbon

Biotin unit)
Tetrahydrofolate:
§ Tetrahydrofolate is derived from vitamin folic acid. It is required in the biosynthesis of purines
Tetrahydrofolate is derived from vitamin folic acid. It is required in the biosynthesis of
and pyrimidines.
purines and pyrimidines.
O COO-

N COO-

H
OH H NH

N
N

H2N N N

Cobalamin (Methyl group transfer)

Tetrahydrofolate

Cobalamin:
§ It is a coenzyme
It iswhich iswhich
a coenzyme derived
is derivedfrom vitamin
from vitamin B12.ofItfour
B . It consists consists ofwith
pyrrole rings
12
four pyrrole rings with Co2+.
2+
Co . Cobalamin participates in the synthesis of acetyl choline. Deficiency of vitamin B12
Cobalamin participates
causes anemia.
in the synthesis of acetyl choline. Deficiency of vitamin B12 causes
anemia.
 Metal
§ Metal ions can also act as ions can also act as cofactors.
cofactors.
Some enzymes and their cofactors are listed below:

Enzymes Metal ion cofactors Function

Cytochrome oxidase Cu2+ Oxidation-reduction

Catalase Fe2+/Fe3+ Oxidation-reduction

Alcohol dehydrogenase Zn2+ Used with NAD+

Carbonic anhydrase Zn2+ Used with NAD+

Carboxypeptidase A Zn2+ Used with NAD+

Arginase Mn2+ removes electrons

Glucose-9-phosphatase Mg2+ Hydrolyzes phosphate esters

Thus some enzymes do not require any cofactor to catalyze the reaction whereas some
enzymes require the presence of metal ions for their catalytic activity. These metal ions play a
crucial role in various enzyme catalytic reaction and they act as catalysts in various ways.
§ Thus some enzymes do not require any cofactor to catalyse the reaction whereas some
enzymes require the presence of metal ions for their catalytic activity.

§ These metal ions play a crucial role in various enzyme catalytic reaction and they act as
catalysts in various ways.