Co-enzymes role in

metabolic pathways
 Introduction
• Coenzymes are small, organic compounds that
are often regarded as cofactors, which are non-
protein compounds that bind to proteins in order
to assist in biochemical transformations.
• They are often vitamins, or vitamin derivatives,
and generally conjugate loosely with the
enzyme’s active site.
• Coenzymes which bind tightly are regarded as
prosthetic groups of the enzyme.
• Coenzymes usually function as activated
carriers, aiding in the transfer of electrons,
specific atoms, or functional groups.
 Coenzymes
• Well-known coenzymes include adenosine triphosphate (ATP), which
transfers phosphate groups; nicotinamide adenine dinucleotide (NADH,
NADPH), which transfers hydrogens and electrons; coenzyme A, which
transfers acetyl groups; and S-adenosylmethionine, which transfers methyl
groups.
• Coenzymes are consumed and recycled continuously in metabolism, with
one set of enzymes adding a chemical group to the coenzyme and another set
removing it.
• For example, enzymes such as ATP synthase continuously phosphorylate
adenosine diphosphate (ADP), converting it into ATP, while enzymes such as
kinases dephosphorylate the ATP and convert it back to ADP.
• Coenzymes molecules are often vitamins or are made from vitamins. Many
coenzymes contain the nucleotide adenosine as part of their structures, such
as ATP, coenzyme A, and NAD+.
 Function of Coenzymes
• Coenzymes work by binding to the active side of the enzymes, the side that
works in the reaction.
• Since enzymes and coenzymes are non-metal organic molecules, they bind
together by forming covalent bonds.
• The coenzymes share electrons with the enzymes, rather than lose or gain
electrons. When they form this bond, they only help the reaction to occur by
carrying and transferring electrons through the reaction.
• Coenzymes do not become integral parts of the enzymatic reaction.
• Instead, the covalent bonds are broken at the end of the reaction, and the
coenzyme returns back to free circulation within the cell until it is used again.
 VITAMINS AND COENZYMES

Vitamin Coenzyme Reaction type Coenzyme class

B 1 (Thiamine) TPP Oxidative decarboxylation Prosthetic group

B 2(Riboflavin) FAD Oxidation/Reduction Prosthetic group

B 3(Pantothenate) CoA - Coenzyme A Acyl group transfer Cosubstrate

B 6(Pyridoxine) Transfer of groups to and from

PLP amino acids Prosthetic group

B 12(Cobalamin) 5-deoxyadenosyl cobalamin Intramolecular rearrangements Prosthetic group

Niacin NAD + Oxidation/Reduction Cosubstrate

Folic acid Tetrahydrofolate One carbon group transfer Prosthetic group

Biotin Biotin Carboxylation Prosthetic group

Table 1 lists vitamins, the coenzymes derived from them, the type of reactions in which they
participate, and the class of coenzyme.
 NAD and NADP
Vitamin B3 (niacin)
• Niacin is the name for both
nicotinamide and nicotinic
acid, either of which can act
as a precursor of
nicotinamide coenzymes.
• Niacin is required for the
synthesis of two coenzyme
molecules: NAD and
NADP.
• The phosphate attached to
the 2 ́- position of the lower
ribose ring in NADP, which
is the only difference
between the molecules.
 NAD+ and NADP+
• NAD + (the oxidized form of the NAD coenzyme) is important in catabolism and
in the production of metabolic energy.
• NADP + (the oxidized form of NADP) is important in the biosynthesis of fats and
sugars.
• NAD + and NADP + play an essential role in many biochemical reactions,
especially redox reactions in which oxidoreductase enzymes transfer hydrogen. 
• (A molecule's acquisition of electrons is called reduction, thus NADH is the
reduced form of NAD + . Conversely, NAD + is the oxidized form, the form with
fewer electrons.) NADP + can be reduced to NADPH, just as NAD + can be
reduced to NADH, although different enzymes will be involved.
• Enzymes that use NAD+ rarely use NADP + and vice versa, making it possible to
separate the biosynthetic and energy-producing functions of NADP + and NAD + .
• NAD+ is often found in conjunction with a "dehydrogenase" enzyme.
 What does NAD do?
• Healthy bodies make all the NADH they need using vitamin B3 as a starting point.
• NAD participates in many redox reactions in cells, including those in glycolysis and
most of the reactions in the citric acid cycle of cellular respiration.
• The electron transport chain in cellular respiration is responsible for energy
production and is an excellent illustration of NAD's involvement in redox reactions. 
• Because of the positive charge on the nitrogen atom in the nicotinamide ring, the
oxidized form of this important redox reagent is often depicted as NAD+.
• In cells, most oxidations are accomplished by the removal of hydrogen atoms.
• NAD coenzyme plays a crucial role in this. Each molecule of NAD+ can acquire two
electrons; that is, be reduced by two electrons. However, only one proton
accompanies the reduction. The other proton produced as two hydrogen atoms are
removed from the molecule being oxidized is liberated into the surrounding
medium. For NAD, the reaction is: 
 FAD / FADH2
• Flavin adenine dinucleotide in its oxidized
state is called FAD. After being reduced, it is
called FADH2.
• The vitamin, riboflavin (or B2) is used to
derive this compound. Riboflavin provides the
ring structures that will directly participate in
the transfer of two hydrogen atoms (each with
one electron this time).
• Similar to NAD, FAD works in association
with a "dehydrogenase" enzyme.
• The reaction removes two hydrogen atoms;
each a proton with one electron. Both
hydrogen atoms bond with FAD.
• This reaction does not release an H+ into
solution like the reduction of NAD does.
 FAD / FADH2
• Flavin adenine dinucleotide in the oxidized form (FAD) accepts two
hydrogen atoms (each with one electron) and becomes FADH2. 
• Similar to NADH, FADH2 will be important as it will deliver hydrogens
and electrons to biochemical processes that can use the electrons and
hydrogens to make ATP.
• The reduced coenzyme FADH2 contributes to oxidative phosphorylation in
the mitochondria.
• FADH2 is reoxidized to FAD, which makes it possible to produce two
moles of the universal energy carrier ATP.
• The source of the energized FADH2 in the cell is generally the TCA cycle.
 Pyridoxal Phosphate (PLP)
• Pyridoxal Phosphate is a coenzyme of many enzymatic reactions. It is the
active form of vitamin B6 which comprises three natural organic
compounds, pyridoxal, pyridoxamine and pyridoxine.
• Pyridoxal phosphate acts as a coenzyme for synthesis of amino acids,
neurotransmitters (serotonin, norepinephrine), sphingolipids,
aminolevulinic acid, transamination reactions, and in some decarboxylation
and deamination reactions of amino acids.
• PLP also is necessary for the enzymatic reaction governing the release of
glucose from glycogen.
• Pyroluria is one potential cause of vitamin B6 deficiency.
 PLP Enzymology

• Central to its mechanism is the formation of a

Schiff base with the enzyme (through a covalent
bond with Lys)
• Schiff base is formed through the condensation
of an amine with a carbonyl group.
• Serves to stabilize carbanion intermediates of
amino acids
• PLP most commonly involved in
decarboxylation or transamination reactions at
the α-Carbon of amino acids
 Thiamine pyrophosphate (TPP)
• Thiamine diphosphate (TPP) is another very important
coenzyme which, like PLP, acts as an electron sink to
stabilize key carbanion intermediates. The important part
of the TPP molecule from a catalytic standpoint is its
thiazole ring.
• Thiamine pyrophosphate is a thiamine (vitamin B1)
derivative which is produced by the enzyme thiamine
diphosphokinase. Thiamine pyrophosphate is
a cofactor that is present in all living systems, in which it
catalyzes several biochemical reactions.
• Thiamine pyrophosphate is synthesized in the cytosol and
is required in the cytosol for the activity of transketolase
and in the mitochondria for the activity of pyruvate-,
oxoglutarate- and branched chain keto acid dehydrogenases
 Chemistry
• Chemically, TPP consists of a pyrimidine ring which is
connected to a thiazole ring, which is in turn
connected to a pyrophosphate (diphosphate) functional
group.
• The part of TPP molecule that is most commonly
involved in reactions is the thiazole ring, which
contains nitrogen and sulfur.
• Thus, the thiazole ring is the "reagent portion" of the
molecule.
 TPP as a coenzyme
• TPP works as a coenzyme in many enzymatic reactions, such as:
1. Pyruvate dehydrogenase complex (AcetylCoA synthesis for acetylcholine)
 pyruvate + CoA --> acetylCoA
2. Alpha-ketoglutarate dehydrogenase complex
 Alpha-ketoglutarate + CoA --> succinylCoA
3. Branched-chain amino acid dehydrogenase complex
 Leucine + CoA 
 Isoleucine + CoA
 Valine + CoA
4. Transketolase
5. 2-hydroxyphytanoyl-CoA lyase
6. Pyruvate decarboxylase in ethanol fermentation
• TPP activates:
 the Krebs cycle (enzymes 1-3) and reduces glucose and pyruvate availability for synthetic pathways
 Transketolase (enzyme 4) that reduces the ribose-5P availability and the nucleotide synthesis
 Biochemical function of TPP
• TPP is a cofactor for enzymes involved in
carbohydrate metabolism, including
transketolase, α-ketoglutarate dehydrogenase,
pyruvate dehydrogenase, and branched chain
α-keto acid dehydrogenase.
• These enzymes are involved in the link
between glycolysis and the citric acid cycle;
the citric acid cycle itself; the pentose-
phosphate pathway; that allow for the
production of ATP, NADPH, and ribose-5-
phosphate which are critical for generating
cellular energy and downstream production
of amino acids, nucleic acids, and fatty acids 
Pathways relying on thiamine diphosphate (TDP)-dependent
enzymes. Enzymes relying on TDP as a cofactor are shown in blue.
 Carboxy Biotin
• Some, although not all, animal carboxylase enzymes (enzymes that add CO 2
to substrates) require the water-soluble vitamin biotin. Biotin is covalently
attached to the enzyme by an amide link to a lysine side chain.
• An ATP-dependent process covalently links CO2 to one of the biotin
nitrogens; the carboxybiotin then acts as a carboxylate donor for the
substrate.
• Biotin functions as a cofactor that aids in the transfer of CO2 groups to
various target macromolecules. Biotin has nine host enzymes with which it
is associated. Humans only have four of these enzymes:
 Biotin dependent enzyme complexes
• Animals have four biotin dependent enzyme complexes:
1. Pyruvate carboxylase, the first step in of the gluconeogenic pathway
from pyruvate, and an important source of oxaloacetate for the TCA
cycle.
2. Acetyl-CoA carboxylase, the control step for fatty acid synthesis (this
enzyme converts acetyl-CoA to malonyl-CoA).
3. Propionyl-CoA carboxylase, which produces methylmalonyl-CoA, the
first step in the conversion of propionyl CoA (generated from odd-chain
fatty acid and some amino acid oxidation) to succinyl-CoA, which can
enter the TCA cycle.
4. β-Methylcrotonyl-CoA carboxylase, an enzyme required for oxidation
of leucine and some isoprene derivatives.
 Biotin deficiency

• Biotin deficiency is sometimes found in consumers of raw chicken eggs,

because raw eggs contain a protein called avidin that binds biotin with very
high affinity and prevents its absorption (avidin is denatured by the cooking
process, so cooked egg consumption is not linked to biotin deficiency).
 Conclusion

• Coenzymes are critical for the metabolism.

• Without these substances, many of the metabolic catalysts that the body
needs to function properly cannot happen. The end result would be the
death of all the cells in the body and therefore the death of the body as well.
• Coenzymes are needed for all phases of metabolism, both anabolism and
catabolism, to help assist enzymes in forming catalytic reactions, which are
necessary for life.