Vitamins and Coenzymes

McMillan Hamwitala (BSc, MSc)

Vitamins
• A group of organic compounds needed in small quantities in the diet
for normal activity of tissues.
• Many vitamins act as cofactors, coenzymes or prosthetic groups for
enzymes
• Most vitamins are derived from diet generally cannot be synthesized
by mammalian cells
• The first vitamin to be discovered was thiamine or B1
• The word Vitamin is derived from the fact that substances
are needed for life (vita) and because thiamine happened to
be an amine
• Not all vitamins are amines or nitrogen containing
compounds
• Vitamin requirements are usually expressed as RDA`s
(recommended dietary allowances)
 Vitamins
Water soluble Lipid soluble
1. Thiamine (B1) 1. Vitamin A
2. Riboflavin (B2) 2. Vitamin D
3. Niacin (B3) 3. Vitamin E
4. Pantothenic acid (B5) 4. Vitamin K
5. Pyridoxal (B6)
6. Biotin (B7)
7. Cobalamin (B12)
8. Folic acid
9. Ascorbic acid (Vitamin C)
Vitamin loss
• Vitamin A: Sensitive to oxygen and light
• Vitamin D: usually little loss
• Vitamin E: sensitive to oxidation especially when heated or with alkali
• Vitamin K: sensitive to acids, alkali, light and oxidizing agents.
• Vitamin C: very sensitive to oxidation, especially when heated in
contact with metals
• Vitamin B complex: water solubility results in loss in cooking water
• Riboflavin is sensitive to light
Cofactors
• Low molecular weight component essential for protein unction
• Metal ions
• Prosthetic groups
• -organic / bioorganic e.g heme groups
• -coenzymes
• Apoenzyme + coenzyme=holoenzyme
Cofactors
• Not all vitamins are cofactors
• All water soluble vitamins with the exception of vitamin C are
converted/activated to cofactors
• Only vitamin K of the fat soluble vitamins is converted to a cofactor
• Cofactors may also act as carriers of specific functional groups such as
methyl groups and acyl groups.
Why cofactors/Coenzymes
• Most enzymes are proteins (or nucleic acids)
• Some chemistry is not easy with amino acid/nucleotide functional
groups
Examples
• Oxygen – binding (hemoglobin)
-C – O bonding –irreversible (physiological T)
Solution – covalent attachment to metal-bound protein
• Chemical capture of light
-photon energy changes covalent structure/denature
Solution – reversible isomerization of cofactor
• Electron transfer reactions
-Protein / NA radicals reactive = denaturation
Poli-aromatic / conjugated co-factors/enzymes like NAD, FAD, quinones…
Classes of coenzymes
• Cosubstrates - are altered during the reaction and regenerated by
another enzyme
• Prosthetic groups – remain bound to the enzyme during the reaction,
and may be covalently or tightly bound to enzyme
• Metabolite coenzymes – synthesized from common metabolites
• Vitamin – derived coenzymes – derivatives of vitamins (Vitamins
cannot be synthesized by mammals, but must be obtained as
nutrients)
Vitamins & coenzymes essentials
• NAD+/NADH
• Most – functional groups / types of chemistry
• Not expected to memorize all structures
• Predict pathologies of deficiencies
Base families
Base families
Example Nucleoside

Nucleobase

2 - deoxyadenosine
Adenosine Triphosphate (ATP)
• Source of immediately usable energy for the cell
• Adenine – containing RNA nucleotide with three phosphate groups
ATP is a versatile reactant that can donate its:
1. Phosphoryl group (ɤ-phosphate)
2. Pyrophosphoryl group (ɤ-β phosphates)
3. Adenylyl group (AMP)
4. Adenosyl group
What requires ATP?
• Transport work
• Mechanical work
• Chemical work
Coenzyme Vitamin Role

ATP Energy and phosphate transfer

NAD(P) Niacin Redox
FAD/FMN Riboflavin (B2) Redox
Coenzyme A Pantothenic acid (B5) Acyl transfer
TPP Thiamine (B1) Transfer of 2 C
PLP Pyrydoxine (B6) Amino acids
Lipoamide Acyl transfer
Ubiquinone Electron carrier
Thiamine (B1)

Heterocyclic components
Pyrimidine (C4N2)
Thiazole (C3SN)
Thiamine (Vitamin B1)
• Active form is thiamine pyrophosphate
• Involved in the oxidative decarboxylation of pyruvic acid and α-
ketoglutaric acid
• Involved in the transketolase reactions of the triose phosphate
pathway
• Also required for nerve function (unrelated to coenzyme activity)
• Lack of thiamine = berberi
Formation of TPP
Pyruvate decarboxylase operates as follows:
Step 1 Nucleophilic attack by the
ylid form of TPP on the carbonyl
carbon
of pyruvate.
Step 2 Departure of CO2 to
generate a resonance-stabilized
carbanion
adduct in which the thiazolium
ring of the coenzyme acts as an
electron sink.
Step 3 Protonation of the
carbanion.
Step 4 Elimination of the TPP ylid
to form acetaldehyde and
regenerate
the active enzyme.
Coenzyme Vitamin Role

ATP Energy and phosphate transfer

NAD(P) Niacin Redox
FAD/FMN Riboflavin (B2) Redox
Coenzyme A Pantothenic acid (B5) Acyl transfer
TPP Thiamine (B1) Transfer of 2 C
PLP Pyrydoxine (B6) Amino acids
Lipoamide Acyl transfer
Ubiquinone Electron carrier
Reactions in which thiamine pyrophosphate is
a cofactor
• Pyruvate decarboxylase
Alcohol fermentation – pyruvate to acetaldehyde
• Pyruvate dehydrogenase
Synthesis of actyl-CoA
• α-ketoglutarate dehydrogenase
Citric acid cycle
• Transketolase reaction
Carbon fixation reactions of photosynthesis
• Acetolactase synthetase
Valine, leucine biosynthesis
Pyridoxal Phosphate (PLP)
• PLP is derived from vitamin B6 family of vitamins
• Vitamin B6 is phosphorylated to form PLP
• PLP is a prosthetic group for enzymes catalyzing reactions involving
amino acid metabolism (isomerization, decarboxylations, side chain
eliminations or replacemements)
Pyridoxine (vitamin B6)
A pyridine derivative
Biochemical functions of vitamin B6
• Pyridoxal phosphate (PLP), the active coenzyme form of vitamin B6 is
closely associated with the metabolism of amino acids.
• Transamination
• Decarboxylation
• Heme synthesis
• Deamination
• Production of niacin
Biotin (Vitamin B7)
• Biotin is required in very small amounts because it is available from
intestinal bacteria
• Avidin (raw egg protein) binds biotin very tightly and may lead to a
biotin deficiency (cooking eggs denatures avidin so it does not biotin)
• Biotin (a prosthetic group) enzymes catalyze:
1. Carboxyl group transfer ractions
2. ATP – dependent carboxylation reactions
• Biotin is linked by an amide bond to the ԑ - amino group of a lysine
residue of the enzyme
• The reactive center of biotin is N-1
Reaction catalyzed by pyruvate carboxylase
• Two step mechanism
• Step 1: formation of carboxybiotin-enzyme complex (requires ATP)
• Step 2: Enolate form of pyruvate attacks the carboxyl group of
carboxybiotin forming oxaloacetate and regenerating biotin
Mechanism of pyruvate carboxylase
The two-phase reaction mechanism of pyruvate carboxylase.
Phase I is a three-step reaction in which carboxyphosphate is formed from bicarbonate and ATP,
followed by the generation of CO2, which then carboxylates biotin.

Phase II is a three-step reaction in which CO2 is produced at the active site via the elimination of
the biotinyl–enzyme, which accepts a proton from pyruvate to generate pyruvate enolate. This
enolate, in turn, nucleophilically attacks the CO2, yielding oxaloacetate.
Pantothenic acid (vitamin B5)

Serves in its activated form as the cofactor for coenzyme A (CoA) and the
Acyl carrier protein (ACP).
Coenzyme A
• Performs a vital role by transporting acetyl groups from one substrate
to another
• The key to this action is the reactive thioester bond in the acetyl form
of CoA
• The thioester bond is stable enough that it can survive inside the cell,
but unstable enough that acetyl-CoA can readily transfer the acetyl
group to another molecule
Example of an acylation reaction

Acetyl-CoA CoA

• Acetylcholine is an important neurotransmitter in the autonomic

nervous system and in the brain
Riboflavin (B2)
• A heterocyclic flavin linked to ribose analogous to the nucleosides in
RNA
• Orange - yellow fluorescent compound
• Found in significant quantities in green leafy vegetables, milk and
meats.
• Heat stable, but easily destroyed by light
• Recommended intake is related to energy intake (kcal) – RDA 1-2
mg/day
RIBOFLAVIN
Dimethylisoalloxazine ring system – confers some
degree of planarity to the molecule and also
color(yellow)

Riboflavin
Riboflavin
• 2 cofactors are involved:
• -riboflavin phosphate (flavin mononucleotide, FMN)
• -flavin adenine dinucleotide (FAD)
• Involved in the metabolism of carbohydrates, fats and proteins (flavin
dehydrogenases/flavoproteins)
• Hydrogen carriers in the respiratory chain
Riboflavin

Reduced substance FAD

dehydrogenase
Cytochrome electron system
(Electron transport chain)

Oxidized substance
FADH2
Riboflavin
Riboflavin
• Enzymes utilizing riboflavin cofactors:
✓NADH dehydrogenase
✓Succinate dehydrogenase
✓D and L amino acid oxidase
✓Pyridoxine-5-phosphate oxidase
✓Glutathione reductase
✓Xanthine oxidase
• In some enzymes, the cofactor is covalently bonded to an amino acid
(dehydrogenases)
Succinate dehydrogenase
Most amino acids (except serine, threonine, ….. And dicarboxylic acids can be deaminated by
L – amino acid oxidases.
 Niacin (vitamin B3

Nicotinic acid
• Two cofactor forms of
niacin: NAD and NADP;
these cofactors are not
tightly held by the
enzyme and may be
reused for reaction after
Nicotinamide reaction.
Coenzyme forms Nicotinamide
Adenine
H O

C
Dinucleotide NH 2
O +
−
N
O P O CH2
O
nicotinamide
H H
H H
OH OH
O NH 2

N
N

−
O P O CH2
N N adenine
O
O H H
H H esterified to
OH OH Pi in NADP+
Biochemical function: NAD+/NADH
H O O
H H
C C
NH2 NH2

+
N + 2 e− + H+ N

R R
NAD+ NADH

The electron transfer reaction may be summarized as

:
NAD+ + 2e− + H+  NADH.
It may also be written as:
NAD+ + 2e− + 2H+  NADH + H+
Vitamin B12

Cyanocobalamin
Ascorbic acid (vitamin C)

Dehydroascorbic acid
Ascorbic acid
• Biochemical functions:
• Production and maintenance of collagen
✓Proline – Hydroxyproline
✓Lysine – hydroxylysine
-Mitochondrial electron transport chain (Cytochrome C)
Folic acid
Fat soluble vitamins
• Vitamins A,D,E and K are the fat soluble vitamins
• Excessive use of vitamins A and K can lead to toxicities
• Fat soluble vitamin tend to be stored in fatty tissues of the body and
in the liver
• Retinol also functions in the synthesis of certain
glycoproteins and mucopolysaccharides necessary
for mucous production and normal growth
regulation
Retinoic acid is important for cellular
differentiation; it controls growth- particularly cell
growth.
Vitamin D
• There are 2 major precursor forms:
7-dehydrocholesterol
Ergosterol
• UV irradiation affords cholecalciferol (vitamin D3) and ergocalciferol
(vitamin D2)
Vitamin D is not a vitamin (or a cofactor) – it is a steroid hormone.
 Photolysis

Non-enzymatic reaction in the skin Transport to the liver

Liver Kidneys

Inactive form