Vitamins

From Essentials of Medical Biochemistry by R.C. Gupta

Vitamins:

A heterogeneous group of organic

compounds

Essential for animals and human beings

Required in very minute quantities

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Vitamins do not provide energy

But their dietary intake is essential

They perform some functions essential for

normal health, growth and reproduction

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Deficiencies of vitamins produce specific
diseases

These can be prevented by intake of pure

vitamins or natural foods containing the
concerned vitamins

Several deficiency diseases were discovered

long before the discovery of the vitamins

In some instances, treatment was discovered

before the discovery of the vitamin

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Scurvy and beriberi are examples

Scurvy was cured by giving citrus fruits even

though the cause of the disease was not known

Beriberi was cured by giving rice polishings

even when its cause was not known

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Since the chemical natures of vitamins were not
known at the time of their discovery, they were
named after the letters of the alphabet

These names have now been largely replaced

by chemical names

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 Vitamins can be classified into
two groups on the basis of their
solubility:

Water-soluble vitamins

Fat-soluble vitamins

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Water-soluble vitamins

Soluble in water

Not stored in the body

Excess intake is wasteful

Excess intake does not cause toxicity

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Water-soluble vitamins include:

Vitamin B-complex family

Vitamin C

EMB-RCG
B-complex family includes:

Thiamin
Riboflavin
Niacin
Pantothenic acid
Pyridoxine
Biotin
Lipoic acid
Folic acid
Cobalamin
EMB-RCG
Since water-soluble vitamins are not stored in
the body, they have to be taken in the diet every
day

Losses can occur during cooking as some of

them are heat-labile

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Some water-soluble vitamins are synthesized
by intestinal bacteria

If intestinal bacteria are destroyed, for example

by antibiotic therapy, additional intake is
required

EMB-RCG
 Thiamin

 Thiamin (vitamin B1) is heat-stable in acidic

medium but not in basic medium

 It is oxidized by mild oxidizing agents to

thiochrome which is biologically inactive

 Chemically, it is made up of a substituted

pyrimidine linked through a methylene bridge to
substituted thiazole
 Functions

 Thiamin forms a coenzyme, thiamin pyro-

phosphate (TPP) or thiamin diphosphate (TDP)

NH2 CH3 O O
| | || ||
C C C — C H 2— C H 2— O — P — O — P — O H
+ | |
N C — C H 2— N OH OH
H 3C — C CH CH S
N
 TPP is a coenzyme for:
• Transketolase
• a-Keto acid dehydrogenases

 a-Keto acid dehydrogenases include:

• Pyruvate dehydrogenase
• a-Ketoglutarate dehydrogenase
• Branched-chain a-keto acid dehydrogenase

 Impaired activity of a-keto acid dehydrogenases

due to thiamin deficiency can limit the availability
of energy
 Sources

 Whole grain cereals, pulses, nuts, yeast, liver,

kidney, heart and meat are good sources of
thiamin

 In cereals, thiamin is present mainly in the outer

layer of the grain

 Removal of the outer layer, e.g. by milling,

causes considerable loss of thiamin
 Requirement

 The recommended daily allowance (RDA) for

thiamin is 0.5 mg/1,000 kcal of energy or 1-1.5
mg/day in adults

 The requirement increases in alcoholics and in

hyper-metabolic states e.g. pregnancy, fever,
hyperthyroidism etc
 Deficiency

 People consuming polished rice or refined

wheat flour as their staple food are susceptible to
thiamin deficiency due to removal of the outer layer
of the grain

 Parboiling of rice decreases the loss of thiamin

 During parboiling, paddy is soaked in warm

water for a few hours and, then, steam-dried
 Thiamin percolates into the deeper part of the
grain

 Polishing of parboiled rice leads to a limited

loss of thiamin

 Alcoholics can develop deficiency as alcohol

impairs the absorption of thiamin and its conversion
into TPP
Deficiency of thiamin causes
beriberi which affects:

Central nervous system

Cardiovascular system

Gastrointestinal tract

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 Central nervous system

 Thiamin deficiency causes peripheral neuritis

involving sensory as well as motor nerves

 Sensory involvement leads to hyperaesthesia,

numbness, tingling and pain

 Motor involvement leads to muscular weakness,

sluggish reflexes, ataxia and paralysis
 Cardiovascular system

 The heart muscle becomes weak resulting in

congestive heart failure

 This, in turn, causes oedema and ascites

 Gastrointestinal tract

 Involvement of gastrointestinal tract causes

anorexia, dyspepsia and constipation
 Predominant involvement of cardiovascular
system is known as wet beriberi because of the
occurrence of oedema

 Predominant involvement of central nervous

system is known as dry beriberi as there is no
collection of water in interstitial tissue in this
condition

 Mixed beriberi is more common in which different

systems are involved in varying degrees
 Diagnosis of beriberi can be confirmed by some
laboratory investigations

 Concentration of pyruvic acid in blood is

increased in beriberi

 Thiamin concentration in erythrocytes is

decreased

 Transketolase activity in erythrocytes is also

decreased
 Urinary thiamin excretion after a test dose is
decreased

 In normal subjects, most of the test dose is

promptly excreted in urine

 Subjects deficient in thiamin retain most of the

test dose in tissues and excrete less in urine
 Riboflavin

 Riboflavin (vitamin B2) is heat-stable in neutral

and acidic medium but not in basic medium

 Its aqueous solution is unstable in sunlight and

ultraviolet light

 Riboflavin can be readily reduced to

leucoriboflavin
 Chemically, riboflavin is 6,7-dimethyl-9-D-ribityl
isoalloxazine

H H H
| | |
C H 2— C — C — C — C H 2O H
| | |
O H O H O H

N N
H 3C — 7 8 9 1
2 O

||
H 3C — 6 3 N H
5 10 4
N
O
 Functions

 Riboflavin is a constituent of flavin

mononucleotide (FMN) and flavin adenine
dinucleotide (FAD)

 These two coenzymes can undergo reversible

oxidation and reduction, and participate in a
number of oxidation-reduction reactions

 The riboflavin portion of FMN and FAD can

reversibly combine with two hydrogen atoms
 H H H O
| | | ||
C H 2— C — C — C — C H 2— O — P — O H
| | | |
O H O H O H O H

N N
H C — O

||
3

H 3 C — N H
N
O
F M N

A H 2

H H H O
| | | ||
C H 2— C — C — C — C H 2— O — P — O H
| | | |
O H O H O H O H
H
N N
H C — O
||
3

H 3 C — N H
N
H O
F M N H 2
FMN is a:

Constituent of respiratory
chain

Constituent of microsomal
hydroxylase system

Coenzyme for L-amino acid

oxidase

EMB-RCG
 H H H O O
| | | || ||
CH—
222 C — C — C — CH— O — P — O — P — O — CH
| | | | |
OH OH OH OH OH NH 2
|
N N N
HC— O N

||
3

FAD
HC—
3
NH
N N
N
O O

H H
H H

OH OH
AH 2

H H H O O
| | | || ||
CH—
222 C — C — C — CH— O — P — O — P — O — CH
| | | | |
OH OH OH OH OH NH 2
H |
N N N
HC— O N
||

FADH 2

HC—
3
NH
N N
N
H
O O

H H
H H

OH OH
FAD is a:

Constituent of respiratory
chain

Constituent of microsomal
hydroxylase system

Coenzyme for many

enzymes

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 Some enzymes requiring FAD as a coenzyme
are:
• D-amino acid oxidase
• Acyl CoA dehydrogenase
• Succinate dehydrogenase
• Glycerol-3-phosphate dehydrogenase
• Xanthine oxidase
• Sphingosine reductase
• Pyruvate dehydrogenase
• a-Ketoglutarate dehydrogenase
Sources
Milk
Eggs
Liver
Kidney
Heart
Yeast
Germinating cereals
Many vegetables
EMB-RCG
 Requirement
 The daily requirement for riboflavin has been
placed at 0.6 mg/1,000 kcal
 Deficiency

 An isolated deficiency of riboflavin is rare

 It is generally combined with other deficiencies

 Clinical features of deficiency are:

• Angular stomatitis (fissures at the angles of

mouth)
• Cheilosis (cracked and swollen lips)
• Glossitis (swollen, painful and magenta-coloured
tongue)
• Seborrheic dermatitis (rough and scaly skin)
• Corneal vascularisation (growth of blood vessels
into the cornea)
 Laboratory diagnosis of riboflavin deficiency is
difficult

 Serum and urinary riboflavin are low in severe

deficiency

 Erythrocyte riboflavin is decreased

 The urinary excretion of riboflavin after a test

dose is decreased
 Niacin

 Niacin was known in the past as anti-pellagra

factor, pellagra-preventing factor and vitamin B3

 It occurs in two forms, niacin (nicotinic acid) and

niacinamide (nicotinamide)

 Both the forms are equally active

 Niacin is converted into niacinamide in the body

 — C O O H — C O N H 2

N N
N ia c in N ia c in a m id e
( n ic o t in ic a c id ) ( n ic o t in a m id e )
 Functions

 Niacin performs its functions in the form of two

coenzymes
• Nicotinamide adenine dinucleotide (NAD)
• Nicotinamide adenine dinucleotide
phosphate (NADP)
 Nicotinamide combines with ribose and
phosphoric acid to form a nucleotide

 This combines with an adenine nucleotide to

form a dinucleotide
 NH 2
|
N
— CO NH 2 N

+
N O O N N
|| ||
O C H 2— O — P — O — P — O — C H 2 O
| |
H H O H O H H H
H H H H

OH O H O H O H*
N A D ( i n N A D P , — O H * i s e s t e r i f ie d w i t h p h o s p h o r i c a c i d )
 These two coenzymes can undergo reversible
oxidation and reduction, and can act as coenzymes
for several oxidoreductases

CH
CH C H 22
CH
CH C — CONH 2 CH C — CONH 2
+
+ H
CH CH CH CH
N + AH 2 A N
| |
R R
+ +
N A D (o r N A D P ) N A D H (o r N A D P H )
 NAD and NADP act as coenzymes in many
metabolic pathways e.g.
•Glycolysis
•Hexose monophosphate shunt
•Citric acid cycle
•Synthesis of fatty acids and steroids
•Oxidation of fatty acids
•Oxidative deamination of amino acids

 Generally, NAD acts as coenzyme in

catabolic pathways and NADP in anabolic
pathways
 Some enzymes which require NAD as a
coenzyme are:
• Glyceraldehyde-3-phosphate dehydrogenase
• Lactate dehydrogenase
• Pyruvate dehydrogenase
• Isocitrate dehydrogenase
• a-Ketoglutarate dehydrogenase
• Malate dehydrogenase
• b-Hydroxyacyl CoA dehydrogenase
• Glutamate dehydrogenase
• IMP dehydrogenase
 NAD is also a constituent of the:
• Respiratory chain
• Microsomal hydroxylase system
 Examples of enzymes requiring NADP as a
coenzyme are:
• Glucose-6-phosphate dehydrogenase
• 6-Phosphogluconate dehydrogenase
• b-Ketoacyl CoA reductase
• a,b-Unsaturated acyl CoA reductase
• Squalene synthetase
• Cholesterol 7-a-hydroxylase
• Thioredoxin reductase
• Haem oxygenase
 Sources

 Milk, eggs, meat, liver, yeast, tomatoes and

green leafy vegetables are good sources of niacin

 Niacin is also synthesized in human beings

from tryptophan

 It has been shown that 1 mg of niacin is

synthesized from 60 mg of tryptophan
 Vitamin B6 is required in the synthetic pathway
as a coenzyme (pyridoxal phosphate)

 Excess of leucine inhibits the conversion of

tryptophan into niacin
 Requirement

 The daily requirement for niacin is 6.6 mg/1,000

kcal

 Or the adult requirement can be taken as 20

mg/day
 Deficiency

 Deficiency of niacin causes pellagra which is

characterized by stomatitis, glossitis, diarrhoea,
dermatitis and dementia (mental degeneration)

 Dermatitis usually affects the exposed parts of

the body

 If untreated, the disease can be fatal

 Pellagra is common in people consuming maize
and sorghum (jowar) as their staple foods

 These two are poor in niacin and tryptophan,

and rich in leucine
 Pantothenic acid

 Pantothenic acid, known in the past as vitamin

B5, is heat-stable in neutral medium but not in acidic
or basic medium

 It is not destroyed by oxidizing or reducing

agents

 It is made up of pantoic acid and b-alanine

Pantoic acid b-Alanine
Pantothenic acid
 Functions

 Pantothenic acid performs its biochemical

functions as a constituent of coenzyme A (CoA) and
acyl carrier protein (ACP)

 Both these compounds contain pantothenic acid

in the form of 4’-phosphopantetheine
 Pantothenic acid is first phosphorylated, by ATP,
at C4 of the pantoic acid residue to form 4’-
phosphopantothenic acid which, then, combines
with cysteine to form 4’-phospho- pantothenyl
cysteine
CH3 O H O H COOH
| || | || | |
C H 2 — C — C H — C — N — C H 2 — C H 2— C — N — C H — C H 2 — S H
| | |
O CH3 OH
|
O = P — OH
|
OH
4 ´ - P h o s p h o p a n to th e n ic a c id C y s te in e
4 ´ - P h o s p h o p a n t o t h e n y l c y s t e in e
 4’-Phosphopantetheine is formed by decarboxy-
lation of the cysteine residue which, after
decarboxylation, is converted into a thio-
ethanolamine residue

 4’-Phosphopantetheine is linked with AMP to

form dephosphocoenzyme A

 The ribose moiety of dephosphocoenzyme A is

phosphorylated at C3 to form coenzyme A
 C H3 O H O H
| || | || |
C H 2— C — C H — C — N — C H 2— C H 2— C — N — C H 2— C H 2— S H
| | |
O C H 3 O H
|
O = P — O H N H2
| |
O N
| N
O = P — O H
|
O
| N N
C H 2

H H
H
H
O O H
|
O = P — O H
|
O
Coenzyme A
 In acyl carrier protein, 4’-phosphopantetheine is
esterified with a serine residue of the protein

 The –SH group of 4’-phosphopantetheine

remains free
 Role of coenzyme A

 Coenzyme A is also represented as CoA-SH as

its terminal –SH group binds various compounds

 This coenzyme participates in a variety of

reactions in the metabolism of carbohydrates, lipids
and amino acids
 Some examples of such reactions are:
• Oxidative decarboxylation of a-keto acids
• Activation of fatty acids
• Activation of some amino acids
 Oxidative decarboxylation of a-keto acids

 A number of coenzymes are required in this

reaction including CoA

 The overall reaction is:

O
||
+
R — C — C O O H + C oA — S H + N A D
 - K e to a c id
O
||
+ +
R — C ~ S — C oA + N A D H + H + C O 2

A cyl C oA
 Pyruvate is converted into acetyl CoA by this
reaction

 a-Ketoglutarate is converted into succinyl CoA

 Activation of fatty acids

 Before fatty acids can take part in any reaction,

they have to be converted into their CoA
derivatives

 This reaction, known as activation of fatty acids,

is catalysed by acyl CoA synthetase (thiokinase)
 Subjects deficient in thiamin retain most of the
test dose in tissues and excrete less in urine
R — C H 2— C O O H + C o A — S H + A T P
F a t t y a c id
 Measurement O of transketolase activity in
||
erythrocytes can confirm the diagnosis
R — C H 2— C ~ S — C o A + A M P + P P i
Acyl C oA
 Activation of amino acids

 Some amino acids, e.g. leucine, isoleucine and

valine, are converted into their CoA derivatives
before they can be metabolised
 An important role of CoA is to provide active
acetate (acetyl CoA) for various reactions e.g.
synthesis of fatty acids, cholesterol, ketone
bodies, acetylcholine etc

 Active succinate (succinyl CoA) is required for

haem synthesis and for gluconeogenesis from
some amino acids
 Role of acyl carrier protein

 Acyl carrier protein is a constituent of the

multienzyme complex which catalyses de novo
synthesis of fatty acids
 Sources

 Pantothenic acid is widely distributed in animal

and plant foods

 It is also synthesized by intestinal bacteria

 Liver, kidney, meat, eggs, yeast, wheat, peas and

sweet potatoes are good sources of pantothenic
acid
 Requirement

 The recommended daily intake is 10 mg

though a smaller intake may be sufficient for
infants and children
 Deficiency

 Deficiency of pantothenic acid has not been

reported in human beings

 In animals, deficiency leads to loss of weight,

loss of hair, greying of hair, anaemia and necrosis
of adrenal glands

 Human deficiency can be produced

experimentally, which causes neurological and
gastrointestinal disturbances
 Pyridoxine

 Pyridoxine was known in the past as vitamin B6

 Vitamin B6 consists of three closely related

pyridine derivatives – pyridoxine, pyridoxal and
pyridoxamine

 All the three are equally active as vitamins

 C H 2O H C HO C H 2N H 2
| | |
H O — — C H 2O H H O — — C H 2O H H O — — C H 2O H

H 3C — H 3C — H 3C —
N N N
P y r id o x in e P y r id o x a l P y r id o x a m in e
 Functions

 Pyridoxine, pyridoxal and pyridoxamine are

phos-phorylated to pyridoxine phosphate, pyridoxal
phosphate and pyridoxamine phosphate
respectively

 The reaction is catalysed by pyridoxal kinase

 The phosphate group is provided by ATP

 Pyridoxine phosphate, pyridoxal phosphate and

pyridoxamine phosphate are interconvertible
 Subjects
CH O H
|
deficient in
2
thiamin retain
CH O H
|
mostO of the 2

||
test dose
HO— — C in
H O tissues
H P and
y r i d o x a lexcrete
2
k in a s e
HO— less in urine — C H 2— O — P — O H
|
+ ATP O H + ADP
H 3C — H 3C —
N N
P y r id o x in e P y r id o x in e p h o s p h a te

 Measurement
CHO
|
of transketolase
CHO
|
activity
O
in
||
erythrocytes
HO— — C H O H can confirm
P y r i d o x a l the
2 H O diagnosis
— — C H 2— O — P — O H
|
k in a s e
+ ATP O H + ADP
H 3C — H 3C —
N N
P y r id o x a l P y r id o x a l p h o s p h a te ( P L P )
C H 2N H 2 C H 2N H 2
| | O
||
HO— — C H 2O H P y r id o x a l HO— — C H 2— O — P — O H
k in a s e |
+ ATP O H + ADP
H 3C — H 3C —
N N
P y r id o x a m in e P y r id o x a m in e p h o s p h a te
 Pyridoxal phosphate and pyridoxamine
phosphate serve as coenzymes, mainly in the
metabolism of amino acids

 Pyridoxal phosphate (PLP) can form a Schiff

base with an a-amino acid

 The union occurs between the a-amino group of

the amino acid and the aldehyde group of
pyridoxal phosphate
 R — C H — C O O H
|
N
||
C — H
|
H O — — C H 2— O — P

H 3C —
N
Schiff base

 The amino acid, thus bound, can undergo

various reactions
 Vitamin B6 coenzymes participate in:
• Transamination
• Deamination
• Decarboxylation
• Transulphuration
• Desulphydration
• Tryptophan metabolism
• Synthesis of haem
• Cellular uptake of amino acids
• Formation of g-amino butyric acid
• Glycogenolysis
 Transamination

 These reactions are catalysed by specific

transaminases

 The amino group of an amino acid is transferred

to an a-keto acid forming a new amino acid and
a new a-keto acid

 PLP acts as a carrier of the amino group

 Transamination reactions are important in :
• Formation of new amino acids
• Catabolism of amino acids
 Subjects deficient in
C H Othiamin retain most of the
|
test dose in tissues and excrete less in urine
HO— — C H 2— O — P
NH2 NH2
| |
R 1— C H — C O O H H 3C — R 2— C H — C O O H
 A m i nMeasurement
o a c id ofN transketolase activity
A m i n o in
a c id
P y r id o x a l p h o s p h a te
erythrocytes can confirm the diagnosis
C H 2N H 2
|
O HO— — C H 2— O — P O
|| ||
R1 — C — CO OH R2 — C — CO O H
H 3C —
 - K e to a c id  - K e to a c id
N
P y r id o x a m in e p h o s p h a te
 Deamination

 PLP acts as a coenzyme for:

• Serine deaminase
• Threonine deaminase
 Decarboxylation

 PLP is a coenzyme for decarboxylases acting on:

• Glutamate
• Arginine
• Tyrosine
 Transulphuration

 PLP is a coenzyme for cystathionine

synthetase and cystathionine g-lyase

 These two transfer sulphur from homocysteine

to serine forming cysteine
 Desulphydration

 PLP is a coenzyme for cysteine desulphydrase

 This enzyme removes the sulphydryl group from
cysteine
 Tryptophan metabolism

 One of the intermediates in the catabolism of

tryptophan is 3-hydroxykynurenine

 This is converted into 3-hydroxyanthranilic acid

by kynureninase, a PLP-dependent enzyme
 When PLP is not available, 3-hydroxyanthranilic
acid is not formed

 3-Hydroxykynurenine is spontaneously converted

into an alternate metabolite, xanthurenic acid

 Xanthurenic acid is excreted in urine

 Urinary excretion of xanthurenic acid can serve

as an indicator of pyridoxine deficiency
 S p o n ta n e o u s
T ry p to p h a n 3 - H y d r o x y k y n u r e n in e X a n t h u r e n ic a c id
PLP K y n u r e n in a s e
3 - H y d r o x y a n t h r a n il ic a c id E x c r e t e d in u r in e

A c e to a c e ty l C o A
 Synthesis of haem

 One of the enzymes involved in the synthesis of

haem is d-aminolevulinic acid synthetase

 This enzyme requires PLP as a coenzyme

 Cellular uptake of amino acids

 Cellular uptake of L-amino acids is an active

process

 This requires the participation of pyridoxal

phosphate
 Formation of g-amino butyric acid

 Gamma-amino butyric acid (GABA) acts as a

neurotransmitter in brain

 It is formed by the action of glutamate

decarboxylase on glutamate

 PLP is required as a coenzyme in this reaction

 Glycogenolysis

 Phosphorylase is a key enzyme of glyco-

genolysis

 PLP acts as a coenzyme for phosphorylase

 Sources

 Liver, eggs, milk, yeast, wheat, corn and green

leafy vegetables are excellent sources of the
vitamin

 Another source is bacterial synthesis in the

intestine
 Requirement

 Since this vitamin is mainly required in the

metabolism of amino acids, its requirement
depends upon the protein intake

 An intake of 1.25 mg/100 gm of proteins has

been recommended
 Deficiency

 Deficiency is very rare

 It may sometimes occur in infants and pregnant

women

 Deficiency may also occur in patients taking

isoniazid, an anti-tuberculosis drug

 Isoniazid forms a complex with pyridoxal and

prevents its activation
 The clinical features of deficiency are nausea,
vomiting, dermatitis, microcytic anaemia and
convulsions

 Convulsions are more common in children

while anaemia is more common in adults

 Chronic deficiency may cause hyperhomo-

cysteinaemia which increases the risk of cardio-
vascular diseases
 Laboratory diagnosis of deficiency can be made
by measuring the urinary excretion of
xanthurenic acid after a test dose of tryptophan

 The urinary excretion of xanthurenic acid is

increased in pyridoxine deficiency
 Biotin

 Biotin is also known as anti-egg white injury

factor

 When raw egg white is fed to rats, they develop

certain symptoms which are relieved by biotin
 It has been shown that raw egg white contains a
protein, avidin

 Avidin forms a complex with biotin preventing its

intestinal absorption

 This leads to a deficiency of biotin

 Avidin is inactivated by heat

 Therefore, cooked eggs do not hamper

absorption of biotin

 Biotin is heat-stable
 Biotin is a heterocyclic, sulphur-containing,
monocarboxylic acid

O
||
C
H N N H
| |
H C C H
| |
H 2C C H — (C H 2)4— C O O H
S
 Functions

 Biotin is a coenzyme for carboxylases

 It is also known as co-carboxylase

 The carboxyl group of biotin forms a bond with

the epsilon-amino group of a lysine residue of the
apoenzyme

 Thus, biotin is firmly bound to the enzyme

 Some carboxylation
reactions requiring biotin:

Carboxylation of
pyruvate

Carboxylation of
acetyl CoA

Carboxylation of
propionyl CoA

EMB-RCG
 Carboxylation of pyruvate

 This reaction converts pyruvate into oxalo-

acetate

 As oxaloacetate is an intermediate in citric acid

cycle, this reaction is important for the normal
operation of citric acid cycle
 C H 3
|
C = O + C O 2 + AT P
|
C O O H
P y ru v a te

B io tin P y r u v a te
c a r b o x y la s e

C O O H
|
C H 2
|
C = O + A D P + P i
|
C O O H
O x a lo a c e ta t e
 Carboxylation of acetyl CoA

 This reaction converts acetyl CoA into malonyl

CoA, and is important in fatty acid synthesis

O
||
C H 3— C ~ S — C o A + C O 2 + ATP
A c e ty l C o A

B io tin A c e ty l C o A
c a r b o x y la s e

C O O H
O
||
C H 2— C ~ S — C o A + A D P + P i
M a lo n y l C o A
 Carboxylation of propionyl CoA

 Propionyl CoA is carboxylated to D-methyl-

malonyl CoA

 This is one of the reactions in the gluconeogenic

pathway for conversion of propionate into glucose
 CH3
|
CH2
|
O = C ~ S — CoA + CO2 + ATP
Propionyl CoA
Propionyl CoA
Biotin carboxylase

CH3
|
H — C — COOH
|
O = C ~ S — CoA + ADP + Pi
D-Methylmalonyl CoA
 Sources

 Bacterial synthesis in the intestine provides

sufficient amounts of biotin

 Dietary sources include egg yolk, liver, kidney,

yeast, peas, tomatoes, cauliflower etc
 Requirement

 Biotin requirement is not known with certainty as

the intestinal bacteria meet most of the
requirement

 The daily intake has been estimated to be 100

to 300 mg
 Deficiency

 Deficiency is unknown in human beings as

suffi- cient biotin is provided by intestinal bacteria,
and biotin is widely distributed in foodstuffs

 Deficiency may occur in animals when they are

fed raw egg white

 It is clinically characterized by retarded growth,

loss of weight, dermatitis, loss of hair, muscular
inco-ordination and paralysis
 Lipoic acid

 Lipoic acid is a sulphur-containing fatty acid

 It is also known as thioctic acid or 6,8-dithio-

octanoic acid

 It can exist in a reduced form and an oxidized

form
C H 2— C H 2— C H — (C H 2)4— C O O H C H 2— C H 2— C H — (C H 2)4— C O O H
| | | |
SH SH A AH S S
2

 - L i p o i c a c id ( r e d u c e d )  - L ip o i c a c id ( o x i d i s e d )
 Functions

 Lipoic acid acts as a coenzyme in some

oxidation-reduction reactions because of its ability
to undergo reversible oxidation and reduction

 It is required as a coenzyme in the oxidative

decarboxylation of a-keto acids such as pyruvate
and a-ketoglutarate
 Sources

 Lipoic acid is widely distributed in foodstuffs

 The quantities required by human beings are

easily obtained from an ordinary diet
 Requirement

 The exact requirement for lipoic acid is not

known

 The vitamin is probably required in very minute

quantities
 Deficiency

 Deficiency of lipoic acid has not been reported

in human beings

 Attempts to induce lipoic acid deficiency in

higher animals have also been unsuccessful
 Folic acid

 Folic acid is also known as folacin or

pteroylglutamic acid

 It is made up of pteridine, para-aminobenzoic

acid and glutamic acid
H 2N N N COOH
1 8 |
2 7 CH2
|
N 3 6 O H CH2
4 5 9 10 || | |
N C H 2— N — — C — N — CH
|
OH | |
H COOH
P t e r id in e p a r a - A m in o - G lu ta m ic
b e n z o ic a c id a c id
P t e r o y lg u t a m ic a c id ( f o lic a c id )
 Folic acid is found in foodstuffs as pteroylmono-
glutamate, pteroyltriglutamate and
pteroylhepta- glutamate

 The last two are converted into pteroylmono-

glutamate in the intestinal mucosa

 Folic acid is heat-stable in neutral medium

 Functions

 Folic acid performs its functions as a coenzyme,

tetrahydrofolate

 Folic acid is first reduced to 7,8-dihydrofolate (H2-

folate or FH2) by dihydrofolate reductase

 7,8-Dihydrofolate is, then, reduced to 5,6,7,8-

tetrahydrofolate (H4-folate or FH4) by dihydro- folate
reductase
 Folate
NADPH + H+
Dihydrofolate reductase
NADP+
Dihydrofolate
NADPH + H+
Dihydrofolate reductase
NADP+
Tetrahydrofolate

 Amethopterin and aminopterin are competitive

inhibitors of dihydrofolate reductase, and act as folic
acid antagonists
 H4-Folate is a carrier of one-carbon units e.g.
• Formyl (–CHO) group
• Formate (–HCOO-) group
• Methyl (–CH3) group
• Methylene (=CH2) group
• Methenyl (=CH) group
• Formimino (–CH=NH) group

 The one-carbon unit may be attached to N5 or N10

of H4-folate
 H
H 2N N N C O O H
|
C H 2
|
N O H C H
5 1 0 || | |
2

| N C H 2— N — — C — N — C H
O H | | |
C H 3 H C O O H
5
N - M e t h y l- H 4 - f o la t e
H
H 2N N N C O O H
|
C H 2
|
N O H C H 2
5 1 0 || | |
| N C H 2— N — — C — N — C H
O H H | |
C H O C O O H
N - F o r m y l- H 4- fo la te ( f10- H 4- fo la te )
1 0

H
H 2N N N C O O H
|
C H 2
|
N O H C H
5 1 0 || | |
2

| N C H 2— N — — C — N — C H
O H | | |
C H H C O O H
||
N H
N 5- F o r m im in o - H 4 - fo la te ( fi5- H 4- fo la te )
 H4-Folate can:
• Receive one-carbon units from various
compounds

• Transfer one-carbon units to various compounds

 Sources of one-carbon units

 Tetrahydrofolate may receive one-carbon units

from:

• Formiminoglutamic acid
• Methionine
• Choline

• Thymine
• Serine
 Formiminoglutamic acid (FIGLU) is formed in
the body from histidine

Histidine Urocanic acid FIGLU

 FIGLU can transfer its formimino group to

tetrahydrofolate

FIGLU + H4-Folate fi5-H4-Folate + Glutamate

 Methionine, choline and thymine are the source
of methyl groups

 Serine can contribute its hydroxymethyl group

 Utilization of one-carbon units

 The one-carbon unit carried by tetrahydrofolate

can be utilized in the following reactions:
• Synthesis of purines
• Synthesis of serine
• Synthesis of methionine
• Synthesis of choline
• Synthesis of thymine
• Synthesis of n-formylmethionine
 Synthesis of purines

 The carbon atoms two and eight of purines are

contributed by f10-H4-folate
 Synthesis of serine

 The hydroxymethyl group for the conversion of

glycine into serine is provided by N5, N10-
methylene-H4-folate
 Synthesis of methionine

 The methyl group for the conversion of

homocysteine into methionine is provided by N 5-
methyl-H4-folate
 Synthesis of choline

 The methyl groups for the synthesis of choline

from serine are provided by N10-methyl-H4-folate
 Synthesis of thymine

 The methyl group of thymine is provided by N5,

N10-methylene-H4-folate
 Synthesis of n-formylmethionine

 The formyl unit of f10-H4-folate converts

methionine into n-formylmethionine which initiates
protein synthesis in prokaryotes
 Sources

 Folic acid is obtained from green leafy

vegetables, yeast, liver, kidney, meat, fish, milk etc

 Intestinal bacteria also synthesize folic acid

 Requirement

Infants and children 100 mg/day

Adult men and women 100 mg/day
Pregnant women 300 mg/day
Lactating women 150 mg/day
 Deficiency

 Folic acid is required for the synthesis of

purines and thymine

 Its deficiency impairs the synthesis of nucleic

acids

 This leads to growth failure and megaloblastic

anaemia

 Leukopenia can also occur

 Laboratory diagnosis of deficiency can be made
by giving a test dose of histidine and measuring the
urinary excretion of FIGLU

 The excretion is increased in subjects deficient in

folic acid
 Cobalamin (Vitamin B12)

 Cyanocobalamin was the first B12 vitamin

isolated as a red crystalline compound from liver in
1948

 Vitamin B12 activity was later found in several

compounds
 Vitamin B12 activity was found in compounds in
which the cyanide group is replaced by:

• Hydroxyl group (hydroxycobalamin)

• Methyl group (methylcobalamin)

• Nitro group (nitrocobalamin)

• Chloride group (chlorocobalamin)

 Vitamin B12 has a complex structure

 Molecular formula of cyanocobalamin is

C63H88O14N14PCo

 It has four pyrrole rings with a cobalt atom at the

centre (corrin ring)

 The tetrapyrrole is heavily reduced and

substituted
 The cobalt atom forms co-ordination bonds
with:
•Nitrogen atoms of four pyrrole rings
•Cyanide group
•5,6-Dimethylbenzimidazole

 5,6-Dimethylbenzimidazole, in turn, is linked

with ribose-3-phosphate

 The phosphate group of ribose-3-phosphate

is linked with the pyrrole ring D (IV) through
 N H 2
|
N H 2 C O
| |
C O C H 2
| |
C H 2 C H 2

H 3C – | |

A C H 3

H 3 C –
N
C N C H 3
H N – O C – H C – |– C H – C O – N H
2 2 2 2

D N C o
+
N B
H N – O C – H 2 C – H 2 C –| – C H 2 – C H 2 – C O – N H 2
|
C H
C H 2 3
N
|
C H – C H 3 C
| C H 3
O |– C H
| | 3

O = P – O H C H 2
C H 3
|
C H 2
|
C O
|
N H 2
N
H 3 C –

H 3 C –
N
C H 2 O H
O

H H
H H
O O H
 Vitamin B12 is heat-stable in acidic and neutral
medium

 It is present in food in association with proteins

 Absorption, transport and storage

 The ingested vitamin B12 is released by gastric

hydrochloric acid

 Most of the vitamin binds to R-protein secreted in

gastric juice and saliva

 Gastric parietal cells also secrete intrinsic factor

(IF), a glycoprotein of 45 kD which can bind
vitamin B12
 However, at low pH, the affinity of vitamin B12 for
R-protein is much higher than that for IF

 Therefore, most of the vitamin binds to R-protein

in stomach

 In the duodenum, R-protein is hydrolysed, and

vitamin B12 is bound to IF
 One IF molecule binds one molecule of vitamin
B12

 A specific receptor on ileal mucosa binds the

IF:vitamin B12 complex

 The vitamin is taken up by the mucosal cells and

is transferred to plasma
 Most of the vitamin is bound to transcobalamin II
in plasma

 The circulating transcobalamin II:vitamin B12

complex is taken up by cells with the help of a
specific receptor

 Transcobalamin II is hydrolysed in the cell by

lysosomal enzymes
 A significant amount of vitamin B12 is stored in the
body, principally in liver

 Most of the vitamin is bound to transcobalamin I

in liver
 Functions

 Vitamin B12 forms coenzymes known as

cobamides which take part in a number of
metabolic reactions

 Cobamides are formed by replacement of the

cyanide group of vitamin B12 by 5’-deoxy-
adenosine
 There are three cobamides:
• 5,6-Dimethylbenzimidazole cobamide: 5,6-Di-
methylbenzimidazole is present as such as
in vitamin B12

• Benzimidazole cobamide: Benzimidazole is

present instead of 5,6-dimethylbenzimidazole

• Adenyl cobamide: 5,6-Dimethylbenzimidazole

is replaced by adenine
 Besides these three, methylcobalamin is active
as a coenzyme as such

 The cobamides function as coenzymes in

various reactions
 Transfer of one-carbon units

 Apart from tetrahydrofolate, cobamides are also

involved in the transfer of one-carbon units

 In fact, they act in concert

 An example of one such reaction is the

synthesis of methionine from homocysteine
 5
N - M e t h y l- H 4 - f o la t e H 4 - F o la t e
C o b a la m in M e t h y lc o b a la m in
M e t h io n in e H o m o c y s t e in e

 H4-Folate is returned to the folate pool so that it

can again participate in one-carbon transfer
reactions
 In cobalamin deficiency, H4-folate can not return
to folate pool and is trapped

 This is known as folate trap

 Thus, cobamides help in one-carbon transfer

reactions by sharing a part of the load on H4- folate
 Formation of succinyl CoA

 Cobamides act as a coenzyme in the conversion

of methylmalonyl CoA into succinyl CoA

 Methylmalonyl CoA is formed from isoleucine,

valine, methionine and fatty acids having an odd
number of carbon atoms

 Succinyl CoA may be converted into glucose or

oxidised in citric acid cycle
 CH3
|
HOOC — C — H
|
C ~ S — CoA
||
O
L - M e th y lm a lo n y l C o A

C o b a m id e M e t h y l m a lo n y l C o A i s o m e r a s e

CH2— COOH
|
C H2 — C ~ S — CoA
||
O
S u c c in y l C o A
 In vitamin B12 deficiency, methylmalonic acid is
excreted in large amounts in urine (methylmalonic
aciduria)

 Rarely, methylmalonic aciduria may be

caused by an inherited defect in methylmalonyl
CoA isomerase
 Sources

 Vitamin B12 can not be synthesized by any plant

or animal

 It is synthesized only by some bacteria

 Animals acquire it through bacterial synthesis in

their intestines
 Liver, kidney, meat, eggs, milk and cheese are
good sources of vitamin B12

 Bacteria present in the human intestine also

synthesize vitamin B12
 Requirement

Age and sex Requirement

Infants and
children 0.3-0.5 µg/day
Adult men and
women 1 µg/day
Pregnant and
lactating women 1.5 µg/day
 Deficiency

 Deficiency of vitamin B12 can occur due to

deficient intake (nutritional deficiency) or due to
impaired absorption

 Absorption is impaired when IF is absent due to

atrophy of gastric mucosa or after gastrectomy

 The condition arising from absence of IF is known

as pernicious anaemia
 Clinical features of deficiency may take long to
develop as the hepatic stores of vitamin B12 can
last a considerable length of time

 Nutritional deficiency causes megaloblastic

anaemia
 An additional feature in pernicious anaemia due
to absence of IF is sub-acute combined
degeneration

 This is degeneration of lateral and posterior

columns of spinal cord leading to sensory as
well as motor disturbances
 Ascorbic acid

 Ascorbic acid (vitamin C) prevents a specific

deficiency disease, scurvy

 Therefore, it is also known as anti-scorbutic

factor

 It is very heat-labile, specially in basic medium

 Chemically, its structure resembles that of

hexoses
 It can exist as L- and D- isomers

 Only the L-isomer possesses vitamin activity

 It can be readily oxidised to dehydroascorbic

acid

 Both L-ascorbic acid and L-dehydroascorbic acid

possess equal vitamin activity
 C = O C = O
| |
C – OH C = O
|| O | O
C – OH C = O
| |
H – C A AH2 H – C
| |
HO – C – H HO – C – H
| |
C H 2O H C H 2O H
L - A s c o r b ic a c i d L - D e h y d r o a s c o r b ic a c id
 Vitamin C is synthesized by all plants and
animals via uronic acid pathway

 The only exceptions are guinea pigs and

primates which require vitamin C from outside

 Guinea pigs and primates lack L-gulonolactone

oxidase, the enzyme that converts L-gulono-
lactone into L-ascorbic acid
 Functions

 Vitamin C is required for the formation and

maintenance of intercellular cement substance

 Since it can undergo reversible oxidation and

reduction, it takes part in some oxidation-
reduction reactions

 There is considerable evidence in support of the

role of vitamin C in the oxidation of
phenylalanine and tyrosine
 It is required for post-translational hydroxylation
of proline and lysine residues

 Therefore, it is essential for the conversion of

procollagen into collagen which is rich in
hydroxyproline and hydroxylysine

 Through collagen synthesis, it plays a role in the

formation of matrix of bones, cartilages, dentine
and connective tissue
 By converting ferric ions into ferrous ions, it
helps in the absorption of iron

 It is required for the formation of bile acids

from cholesterol

 It acts as an anti-oxidant

 It inhibits the formation of nitrosamines

(carcinogens) during digestion
 Ascorbic acid may play a role in the synthesis of
adrenocortical steroid hormones

 Ascorbic acid content of adrenal gland is found

to decrease rapidly in stressful conditions
 Tissue distribution

 Total amount of ascorbic acid in an adult is 2-3

gm

 It is distributed in all tissues and body fluids

 It is present in high concentrations in the

glands
 The highest concentration is found in the
adrenal glands, followed by the pituitary gland,
thymus and corpus luteum

 Leukocytes are also rich in ascorbic acid

 The concentration in plasma is 0.5 -1.5 mg/dl

 The vitamin begins to appear in urine when the

plasma level exceeds 1 mg/dl
 Sources

 Citrus fruits, e.g. amla, lemon and orange, are

very rich in vitamin C

 Guava, tomatoes, green leafy vegetables and

germinating pulses are also good sources
 Potatoes are a fair source

 Since considerable losses of vitamin C can

occur during cooking, some raw fruits and salads
should be included in the daily diet
 Requirement

 Vitamin C is required in much larger

quantities than any other vitamin

 The RDA according to Indian Council of Medical

Research is :

Infants and children 30-50 mg/day

Adult men and women 60 mg/day
Pregnant and
lactating women 80 mg/day
 Deficiency

 Deficiency of vitamin C produces scurvy

 A full-blown picture of scurvy is rare these days

but isolated signs and symptoms of vitamin C
deficiency are still seen
 The signs and symptoms include:
• Swollen, spongy and bleeding gums
• Loosening of teeth
• Petechial haemorrhages
• Anaemia
• Retardation of skeletal growth
• Easy fracturability of bones
• Delayed union of fractures
• Delayed healing of wounds
• Hypercholesterolaemia
Petechial haemorrhages
 Laboratory diagnosis of deficiency can be made
by ascorbic acid saturation test

 After a test dose of ascorbic acid, urinary

excretion of ascorbic acid is low in subjects
deficient in the vitamin
 Fat-soluble vitamins

 These are soluble in fat but insoluble in water,

and include vitamins A, D, E and K

 They are present in food in association with

dietary lipids

 They can be stored in the body and their

excessive intake can be toxic
 Vitamin A

 Vitamin A is found only in animals though its

precursors are found in a variety of plants

 It occurs in three forms, retinol (alcohol form),

retinal (aldehyde form) and retinoic acid (acid form)

 Chemically, each form contains a b-ionone ring

attached to a polyene chain
 H C H H C H
C H |
3 3

|
3
| | |
C C C C C H 2O H
8 10 12 13 14 15
5 7 C 9 C 11 C
4 6 C R e t in o l

3 1 H H H H
C H 3
2
C H 3

H C H H C H
C H |
3 3

|
3
| | |
C C C C C HO
C C C C R e t in a l
H H H H
C H 3

C H 3

H C H H C H
C H |
3
| |
3

|
3
|
C C C C C O O H
C C C C R e t in o i c a c i d
H H H H
C H 3

C H 3
 All the double bonds in the polyene chain have a
trans-configuration in naturally occurring vitamin A
(all-trans-vitamin A)

 Retinol may be esterified with a fatty acid or

phosphoric acid

 The storage form of vitamin A is generally retinyl

palmitate

 Some of the functions of vitamin A are performed

by retinyl phosphate
 Vitamin A can be formed in our body from its
precursors (provitamins A)

 The precursors are carotenes (or carotenoids)

which are naturally occurring pigments found in
most yellow and green fruits and vegetables
 a-Carotene, b-carotene and g-carotene are
important precursors of vitamin A

 Of these, b -carotene is the most important

 One molecule of b -carotene can be converted

into two molecules of retinal
 A molecule of b-carotene contains two b-ionone
rings connected by an 18-carbon polyene chain

 b-Carotene is cleaved in the middle by b -

carotene dioxygenase and molecular oxygen to
form two molecules of retinal

 Bile salts facilitate the reaction

 H 3C
C H H C H 3 H C H 3 H H H H H H 3C
3
| | | | | | | | |
|
C C C C C C C C C
C C C C C C C C C |
| | | | | | | | |
C H 3H H H H H C H H C H 3 H C H 3
3
C H 3
 -C a ro te n e

[O 2]  - C a r o te n e d io x y g e n a s e ,
b ile s a lts
H 3C
C H H C H 3 H C H 3 H H H H H C
3
| | | | | | | | 3
|
C C C C C H O C C C C
C C C C + O H C C C C C
| | | | | | | | |
C H 3H H H H C H 3 H C H 3 H C H 3
C H 3 R e t in a l R e t in a l
+
N AD PH + H
R e t in a l d e h y d e r e d u c t a s e [O ]
( r e tin e n e r e d u c ta s e ) S p o n ta n e o u s
+
N AD P
H C H H C H H C H H C H
C H |
3
| |
3
C H | |
3
| |
3

|
3
| |
3

C C C C C H 2O H C C C C C O O H
C C C C C C C C
| | | | | | | |
C H 3H H H H C H 3H H H H
C H 3 C H 3
R e t in o l R e t in o i c a c id
 Retinal is reduced to retinol by retinaldehyde
(retinine) reductase

 Some retinal is spontaneously oxidised to

retinoic acid

 While retinal and retinol are interconvertible,

retinoic acid cannot be converted back into retinal
 a-Carotene and g-carotene contain only one
b- ionone ring

 Therefore, they can form only one retinal

molecule each
 Absorption, transport and storage

 Carotenes are converted into retinal and, then,

into retinol in the small intestine

 A small amount of carotenes may be absorbed

as such and transported by chylomicrons

 Retinol is absorbed from the small intestine

 Retinyl esters are hydrolysed to form retinol

 Retinol is taken up by the mucosal cells, and is
re-esterified with saturated fatty acids

 Esterified retinol enters the lacteals and

reaches liver via circulation
 Retinol is stored in the hepatic lipocytes

 It is released from hepatic cells into circulation

 It is transported in blood by an a1-globulin,

retinol binding protein (RBP)
 Circulating retinol is taken up by various cells in
which it is bound to cellular retinol binding protein
(CRBP)

 Retinoic acid is transported in circulation in

association with albumin
 Functions
 Functions of vitamin A are related to:
• Reproduction
• Growth and differentiation
• Integrity of epithelial tissues
• Vision
• Anti-oxidant role
• Anti-carcinogenic role
 Reproduction

 Vitamin A deficiency has been observed to impair

reproductive function in some animals

 This function of vitamin A is probably mediated

by control of expression of certain genes by retinol
bound to CRBP
 Growth and differentiation

 Vitamin A is required for normal growth and

differentiation of tissues

 Both skeletal growth and formation of soft

tissues are impaired in growing animals deficient in
vitamin A

 Vitamin A seems to be essential for the

synthesis of mucopolysaccharides and
glycoproteins in various tissues
 Integrity of epithelial tissues

 Vitamin A maintains the integrity of epithelial

tissues

 The epithelial tissues of eyes, lungs,

gastro- intestinal tract and genitourinary tract
become dry and keratinised in vitamin A deficiency
 Vision

 Role of vitamin A in vision was demonstrated by

Morton and Wald

 Retina contains two types of photoreceptor

cells, rods and cones

 Rod cells are required for dim light vision and

cone cells for bright light vision including the
perception of colours
 Vitamin A is essential for the functioning of rod
cells as well as cone cells

 Rod cells contain a pigment, rhodopsin (visual

purple) which is a conjugated protein made up of
opsin (a protein) and 11-cis-retinal, an isomer of all-
trans-retinal
 The aldehyde group of 11-cis-retinal is bonded
with the epsilon-amino group of a lysine residue of
opsin by Schiff base linkage

 Rhodopsin molecules are present in large

numbers in rod cells

 When light strikes rod cells, 11-cis-retinal is

converted into all-trans-retinal
 Conformation of all-trans-retinal is such that it can
not bind with opsin

 Therefore, opsin and all-trans-retinal dissociate

 A number of transient intermediates are formed

before dissociation of opsin and all-trans-retinal
 When a person moves from dim light into bright
light, all the rhodopsin molecules are rapidly
dissociated

 When he moves back into dim light, he is

unable to see until rhodopsin is re-formed

 For this, all-trans-retinal has to be isomerised to

11-cis-retinal
 The enzyme system for the isomerisation
reaction is present only in liver

 Therefore, all-trans-retinal is reduced to all-

trans-retinol in retina, and is released into circulation

 It is taken up by liver, and is converted into 11-

cis-retinol which is released into circulation
 Retina takes up the circulating 11-cis-retinol and
oxidises it to 11-cis-retinal

 The latter combines with opsin to form rhodospin

 This sequence of reactions is known as visual

cycle
 L ig h t L ig h t
R h o d o p s in B a th o r h o d o p s in L u m ir h o d o p s in
L ig h t

M e ta r h o d o p s in I
L ig h t

M e ta r h o d o p s in II R E T IN A
L ig h t
O p s in
II- c is - R e tin a l A l l - t r a n s - r e t in a l
+
NADPH + H
R e t in i n e r e d u c t a s e
+
NADP
II- c is - R e tin o l A l l - t r a n s - r e t in o l
v ia v ia
c i r c u la t i o n c i r c u la t i o n
R e t in o l i s o m e r a s e
II- c is - R e tin o l A l l - t r a n s - r e t in o l L IV E R
 The time taken for regeneration of rhodospin is
known as the dark adaptation time which
depends upon the vitamin A content of liver

 When liver contains adequate stores of vitamin

A, the dark adaptation time is short

 In vitamin A deficiency, dark adaptation time is

prolonged
 Mechanism of vision

 Rhodopsin is a transmembrane protein

 When photons strike it, they cause a change in

its conformation

 As a result, rhodopsin molecule is activated

(photo-excited)

 The photo-excited form is probably meta-

rhodopsin II
 Another membrane-bound protein, transducin
is present in close vicinity of rhodopsin

 Transducin belongs to the family of G-proteins

 It is made up of three subunits, a-subunit, b-

subunit and g-subunit

 The a-subunit has a site for GDP or GTP

 Normally, this site is occupied by GDP

 Photo-excited rhodopsin causes displacement
of GDP by GTP

 The a-subunit containing GTP dissociates from

the b- and g-subunits

 It binds to and activates the enzyme, cGMP

phospho-diesterase

 The active enzyme converts cGMP into G-5’-MP

 Decrease in concentration of cGMP causes
hyperpolarisation of the rod cell membrane

 The rod cell membrane contains a number of

cation-specific channels

 These are normally kept open by cGMP

 Open channels lead to continuous influx of

sodium ions into the rod cell
 Decrease in cGMP concentration leads to
closure of cation-specific channels

 Sodium ions accumulate outside the rod cell

 This causes hyperpolarisation of the rod cell

membrane
 Light Photo-excited
Rhodopsin
rhodopsin

GDP-transducin GTP-transducin

GTP-a-subunit b- and g-subunit

Inactive cGMP Active cGMP

phosphodiesterase phosphodiesterase

cGMP GMP

Open cation-specific Closed cation-specific

channels channels

Normal Hyperpolarised
membrane membrane
 The nerve impulse generated as a result of
hyperpolarisation is transmitted to appropriate
areas of brain (visual cortex)

 The a-subunit of transducin possesses intrinsic

GTPase activity

 Therefore, GTP bound to the a-subunit is slowly

hydrolysed into GDP
 When GTP is converted into GDP, the a-subunit
re-associates with the b- and g-subunits
forming transducin containing GDP

 cGMP phosphodiesterase becomes inactive

again, and the whole chain of events is terminated
 Vitamin A is also required for bright light vision

 The cone cells of retina contain three

conjugated proteins, porphyrinopsin, iodopsin
and cyanopsin

 These are sensitive to red, green and blue light

respectively

 The prosthetic group is 11-cis-retinal in each of

these but the protein part is different
 These proteins become photo-excited on
exposure to light of specific colour

 A nerve impulse is, then, generated in much the

same way as in rod cells

 A given cone cell contains only one of these

three proteins, and can perceive only one particular
colour

 An inherited absence of, or defects in, these

proteins causes colour blindness
 Anti-oxidant role

 b-Carotene acts as an anti-oxidant

 It prevents lipid peroxidation at low oxygen

tension
 Anti-carcinogenic role

 Some epidemiological studies have shown that a

low intake of vitamin A or carotenes is associated
with a high incidence of certain cancers

 However, experimental and clinical confirmation

of this finding is yet to be obtained
 Since retinol and retinal are interconvertable and
can also be converted into retinoic acid, they can
perform all the functions of vitamin A

 Retinoic acid can support growth and

differentiation, and can maintain epithelia but
cannot perform other functions of vitamin A

 13-cis-Retinoic acid, a synthetic compound, is

more active than naturally occurring retinoic acid
 Sources

 Preformed vitamin A is present only in animal

foods e.g.
• Fish liver oils
• Liver
• Eggs
• Milk
• Butter
• Cream
• Cheese
 Provitamins A are found in:
• Green leafy vegetables e.g. spinach,
coriander leaves, mustard leaves, cabbage,
lettuce

• Red and yellow vegetables and fruits e.g.

carrot, tomato, mango, papaya, plums
 Requirement

 Vitamin A is present in food in several different

forms

 These forms differ in their vitamin A activity

 Therefore, it is not possible to express the

requirement for vitamin A in terms of its actual weight
 To overcome this difficulty, relative vitamin A
activities of different compounds were compared in
rats

 The activities were expressed in terms of

international units (IU)

 One IU is the activity present in 0.3 µg of retinol,

0.344 µg of retinyl acetate or 0.6 µg of
b-carotene
 Daily requirements were expressed in IU for a
long time

 Later, it was found that absorption and utilisation

of carotenes were much less efficient in human
beings than in rats

 Therefore, vitamin A activities were converted into

retinol equivalents (RE)

 One RE is the activity present in 1 µg of retinol,

6 µg of b-carotene or 12 µg of other carotenes
The daily requirement of vitamin A in terms of
IU and RE is:

IU/day RE/day
Infants 1,500-2,000 400
Children 2,000-4,000 400-700
Adult men 5,000 1,000
Adult women 4,000 800
Pregnant women 5,000 1,000
Lactating women 6,000 1,200
 Deficiency

 Deficiency of vitamin A can arise from:

• Inadequate intake
• Inadequate absorption
• Inadequate conversion of carotenes into retinal

 Zinc deficiency may impair the mobilisation of

vitamin A from liver
 The clinical manifestations of deficiency are:
• Xerophthalmia
• Keratomalacia
• Nyctalopia
• Blindness
• Follicular hyperkeratosis
• Susceptibility to infections
 Xerophthalmia

 The lacrimal glands become keratinised, and

stop secreting lacrimal fluid

 The eyes become dry

 Bitot’s spots (small, opaque spots) may appear

on cornea
Bitot’s spot
 Keratomalacia
 The cornea is softened

 It is finally destroyed
 Nyctalopia

 Initially, dark adaptation time is prolonged which

may be the earliest indication of deficiency

 Finally, the ability to see in dim light is

completely lost which is known as night blindness
or nyctalopia
 Blindness

 In severe and prolonged deficiency, there is

total loss of vision due to functional and structural
changes in the eyes
 Follicular hyperkeratosis

 There is hyperkeratinisation of skin especially

around hair follicles
 Susceptibility to infections

 Susceptibility to infections may be increased

due to epithelial damage
 Vitamin A toxicity

 Hypervitaminosis A may occur if large doses of

vitamin A, usually in pharmacological form, are
taken over long periods

 It is characterized by rough skin, hair loss,

anorexia, weight loss, headache, vertigo,
irritability, hyperaesthesia, hepatosplenomegaly,
liver damage etc
 Vitamin D

 Vitamin D prevents a deficiency disease, rickets

(rachitis), and is also known as:
• Anti-rachitic factor
• Anti-rachitic vitamin

 Of the several compounds possessing

vitamin D activity, the most important are:
• Vitamin D2 (ergocalciferol)
• Vitamin D3 (cholecalciferol)
 Vitamin D2 is formed from provitamin D2
(ergosterol) on exposure to ultraviolet light

 Vitamin D3 is formed from provitamin D3

(7-dehydrocholesterol) on exposure to ultraviolet
light
 H 3C H 3C

C H C H
C H 3
3
C H 3
3

C H 3 C H 3
C H 3 C H 3

C H 3
C H 2

U lt r a v io le t lig h t

H O H O
E r g o s te ro l E r g o c a lc if e r o l ( v i ta m i n D 2 )

H 3C H 3C

C H C H
C H 3
3
C H 3
3

C H 3 C H 3

C H 3
C H 2

U lt r a v io le t lig h t

H O H O
7 - D e h y d r o c h o le s te r o l C h o le c a l c i f e r o l ( v it a m in D 3 )
 Ergosterol occurs in plants e.g. ergot and yeast

 7-Dehydrocholesterol occurs in animals

 7-Dehydrocholesterol is formed from cholesterol

 7-Dehydrocholesterol present in skin is conver-

ted into cholecalciferol on exposure to sunlight

 Cholecalciferol is synthesized in human beings

also
Functions

Intestinal absorption of calcium and

phosphorus

Renal tubular absorption of calcium

Mineralization of bones

EMB-RCG
 Vitamin D increases the intestinal absorption
of calcium

 The absorption of phosphorus is also increased

secondarily

 Vitamin D increases the renal tubular absorption

of calcium, and decreases that of phosphorus
 Vitamin D is required for mineralization of
bones

 Bone formation is a continuous and

dynamic process

 Calcium salts are continuously deposited into

and resorbed from bones by osteoblasts and
osteoclasts respectively
 In growing age, deposition exceeds resorption
leading to skeletal growth

 In adult life, deposition and resorption are finely

balanced leading to remodeling of bones

 Vitamin D is required for bone growth as well as

remodeling
 Thus, the main functions of vitamin D are to
regulate the metabolism of calcium and the
mineralization of bones

 However, these functions are not performed by

vitamin D itself

 Vitamin D is first hydroxylated at C25 to form

25-hydroxycholecalciferol in the liver
 25-Hydroxycholecalciferol is further
hydroxylated at C1 in the kidneys to form 1,25-
dihydroxy- cholecalciferol (1,25-DHCC or calcitriol)

 1,25-DHCC is the metabolically active form of

vitamin D

 1,25-DHCC acts as a hormone and, therefore,

vitamin D may be regarded as a prohormone
 The mechanism of action of 1,25-DHCC is
similar to that of steroid and thyroid hormones

 It binds to its intracellular receptors in the target

cells

 The hormone-receptor complex binds to a

hormone response element in DNA

 This increases the transcription and translation

of certain genes
 C h o le c a l c i f e r o l

L IV E R H y d r o x y la s e

2 5 - H y d r o x y c h o le c a lc ife r o l

K ID N E Y H y d r o x y la s e
+ P a r a th o rm o n e P a r a t h y r o id
g la n d s
1 , 2 5 - D ih y d r o x y c h o le c a lc ife r o l
IN T E S T IN A L
M UCO SA In d u c tio n

C a lc i u m - b i n d i n g p r o t e i n –
C a ++ -d e p e n d e n t A T P a s e
A lk a lin e p h o s p h a ta s e

IN T E S T IN A L
M UCO SA

 C a lc i u m a b s o r p t i o n R e le a s e i n t o c i r c u la t io n  P la s m a c a lc iu m
 Sources

 Plant foods are poor sources of vitamin D

 Though ergocalciferol is present in some plants,

its intestinal absorption is poor

 Cholecalciferol is the major dietary form of

vitamin D

 Fish liver oils are very rich in cholecalciferol

 Eggs, butter and cheese are fairly good sources

 An important source of vitamin D is its

endogenous synthesis in the skin

 Where sunshine is good and people are

adequately exposed to sunlight, enough vitamin D
is synthesized in the body to meet the daily
requirement
 Requirement

 Due to differences in the anti-rachitic activity of

different forms of vitamin D, the requirement is
expressed in international units (IU)

 One IU is the activity present in 0.025 µg of

cholecalciferol
 The requirement is 400 IU/day in infants,
children, and pregnant and lactating women

 The adult requirement is 200 IU/day

 Deficiency

 Deficiency can occur due to:

• Inadequate intake
• Inadequate absorption
• Inadequate exposure to sunlight

 Two distinct syndromes arise from deficiency of

vitamin D:
• Rickets in childhood
• Osteomalacia in adult life
 Rickets

 The most susceptible periods for development of

rickets are infancy and puberty when skeletal
growth occurs rapidly

 Deficiency of vitamin D causes deficient

mineralization of bones and overgrowth of
epiphyses

 This results in some typical skeletal deformities

 Delayed closure of fontanelles

 Closure of fontanelles is delayed in rickets

 It causes bossing of skull

 Craniotabes

 The skull bones are soft due to poor

mineralization

 This is known as craniotabes

 Pigeon chest

 The antero-posterior diameter of chest is

increased

 This gives the appearance of pigeon chest

 Rickety rosary

 The costochondral junctions of ribs are

enlarged, and appear to the beaded

 This series of beads on the chest gives

the appearance of a rosary worn around the neck

Enlarged
costochondral
junctions
 Vertebral deformities

 Kyphosis, lordosis or scoliosis can occur

 Kyphosis is forward bending of vertebral column

 Lordosis is backward bending of vertebral

column

 Scoliosis is lateral bending of vertebral column

 Pelvic deformities

 Pelvis may be deformed

 The size of the pelvic outlet may be

decreased
 Bowlegs and knock knees

 The leg bones may become

curved under the weight of the
body (bowlegs)

 The two knees may touch

each other while standing
erect (knock knees)
 Enlarged wrists and knees

 Wrists and knees may be enlarged due to

overgrowth of epiphyses of the long bones
Multiple skeletal
deformities may
cause shortening
of stature
 Apart from the skeletal deformities, some
changes in serum and urine chemistry are also
seen

 Serum calcium and inorganic phosphorus are

decreased

 Serum alkaline phosphatase is increased

 Hypocalcaemia increases the secretion of
parathormone

 Urinary excretion of calcium is decreased,

and that of phosphorus is increased by
parathormone
 Osteomalacia

 Pregnant and lactating women are particularly

prone to osteomalacia

 Demineralization of bones occurs to varying

extents

 Aches and pains may occur in bones, and

susceptibility to fractures is increased

 Changes in serum and urine chemistry are similar

to those in rickets
 Toxicity

 Prolonged intake of large doses of vitamin D in

pharmacological form may cause toxicity (hyper-
vitaminosis D)

 Serum calcium is raised

 Hypercalcaemia may cause loss of appetite,

polydipsia, polyuria, constipation, muscular
weakness etc
 Calcification may occur in soft tissues e.g.
arteries, bronchi, muscles, kidneys etc

 Stones may be formed in the kidneys

 Vitamin E

 Vitamin E is sometimes described as anti-sterility

vitamin

 However, its anti-sterility function is seen only in

some animals and not in human beings

 Vitamin E activity is present in several

tocopherols, the most important being a-, b-, g- and
d- tocopherols
 C H 3
|
H O — 5 4
6 3
7 2 C H 3 C H 3
H 3C — 8 1 | |
O (C H 2)3— C H — (C H 2)3— C H — (C H 2)3— C H — ( C H 3)2
|
C H 3 C H 3  -T o c o p h e ro l
C H 3
|
H O —

C H 3 C H 3
| |
O (C H 2)3— C H — (C H 2)3— C H — (C H 2)3 — C H — ( C H 3)2
|
C H 3 C H 3
-T o c o p h e ro l

H O —

C H 3 C H 3
H 3C — | |
O (C H 2)3— C H — (C H 2)3— C H — (C H 2)3 — C H — ( C H 3)2
|
C H 3 C H 3 -T o c o p h e ro l

H O —

C H 3 C H 3
| |
O (C H 2)3— C H — (C H 2)3— C H — (C H 2)3 — C H — ( C H 3)2
|
C H 3 C H 3
-T o c o p h e ro l
 a-Tocopherol is the most abundant tocopherol
in foods, and is taken as the standard

 The official reference standard is synthetic DL-

a- tocopherol acetate

 Vitamin E activity present in 1 mg of this

compound is taken as one international unit (IU)
 The vitamin E activity of 1 mg of naturally-
occurring D-a-tocopherol is 1.5 IU

 The vitamin E activities of other tocopherols are

much lower
 Functions

 The most important function of vitamin E in

human beings is to act as an anti-oxidant

 Vitamin E is readily oxidisable, and prevents the

oxidation of other, less oxidisable compounds

 It prevents the oxidation of other antioxidants

e.g. carotenes, vitamin A and vitamin C
 It prevents peroxidation of unsaturated fatty
acids, and protects the tissues against the harmful
effects of lipid peroxides

 It protects the RBC membrane from oxidants,

and makes the RBCs resistant to haemolysis

 It protects the pulmonary tissue from

atmospheric oxidants
 Sources

 Vegetable oils, e.g. wheat germ oil, rice bran oil,

corn oil, soya bean oil, cottonseed oil etc, are very
rich in vitamin E

 Other good sources are green leafy vegetables,

nuts, legumes, milk, eggs and meat
 Requirement

Infants 4-5 IU/day

Children 7-12 IU/day
Adult men 15 IU/day
Adult women 12 IU/day
Pregnant and
lactating women 15 IU/day
 The requirement of vitamin E is related to the
intake of polyunsaturated fatty acids (PUFA)

 It has been suggested that an intake of 0.8 mg

of D-a-tocopherol (or 1.2 IU of vitamin E) per gm
of PUFA in diet will prevent vitamin E deficiency
 Deficiency

 Deficiency of vitamin E is uncommon because of

its widespread distribution in foods

 Moreover, the foods which are rich in PUFA, e.g.

vegetable oils, are also rich in vitamin E
 Deficiency may occur in severely
undernourished children and in premature
infants fed on artificial milk not containing vitamin
E

 The clinical manifestations of deficiency are

oedema, haemolytic anaemia and
thrombocytosis
 Vitamin K

 Vitamin K activity is present in many

compounds having a 2-methyl-1,4-
naphthoquinone nucleus

 Two such natural compounds are

phylloquinone (vitamin K1) and menaquinone
(vitamin K2)

 The former occurs in plants and the latter in

bacteria
O

1
2 — CH 3
CH3 CH3 CH3
| | |
3 — C H 2 — C H = C — ( C H 2) 3 — C H — ( C H 2) 3 — C H — ( C H 2) 3 — C H — ( C H 3 ) 2
4

O
2 - M e t h y l - 3 - p h y t y l - 1 , 4 - n a p h t h o q u i n o n e (Phylloquinone)
O

— CH 3
CH3
|
— C H 2— ( C H = C — C H 2) 5— C H = C — ( C H 3) 2

O 2 - M e t h y l - 3 - d i f a r n e s y l - 1 , 4 - n a p h t h o q u i n o n e (Menaquinone)
 Some synthetic compounds having vitamin K
activity have also been prepared

 These include menadione, sodium menadiol

diphosphate and menadione sodium bisulphite

 These three are water-soluble

 ONa
|
O — P = O
O O |
ONa
S O 3N a
— CH 3 — CH3 — CH 3

ONa
O O |
M e n a d io n e M e n a d i o n e s o d i u m b is u lp h i t e
O — P = O
|
ONa
S o d iu m m e n a d io l d ip h o s p h a te
 Functions

 Vitamin K is essential for normal coagulation of

blood

 Concentrations of several coagulation factors in

plasma are decreased in vitamin K deficiency

 These include prothrombin (Factor II),

proconvertin (Factor VII), Christmas factor (Factor
IX) and Stuart-Prower Factor (Factor X)
 All these coagulation factors are proteins

 They require some post-translational modification

for their activation

 Vitamin K is required for the post-translational

modification
 Prothrombin is synthesized in liver in the form of
an inactive precursor, pre-prothrombin

 Pre-prothrombin is converted into prothrombin

by carboxylation of its glutamate residues

 Hydroquinone (reduced) form of vitamin K is

required for the carboxylation reaction

 During this reaction, hydroquinone form of

vitamin K is converted into 2,3-epoxide form
 Hydroquinone form is regenerated via quinone
form by 2,3-epoxide reductase and vitamin K
reductase

 An as yet unidentified sulphydryl compound is

required in the reaction catalysed by 2,3-epoxide
reductase
 C O O H H O O C C O O H
|
C H 2 C H
| |
H C H 2 O H C H 2 O
| | || | | ||
— N — C H — C — — N — C H — C —
G lu ta m a te r e s id u e -C a rb o x y g lu ta m a te r e s id u e
V ita m in K - d e p e n d e n t
c a r b o x y la s e
C O 2 + O 2 H 2O

O H O
| ||
— C H 3 — C H 3

O
— R — R
| ||
O H O
V ita m in K V ita m in K
( h y d r o q u in o n e fo r m ) ( 2 ,3 - e p o x id e fo r m )
S H
N A D P +
R
V ita m in K S H
re d u c ta s e S
R + H 2O  2 , 3 - E p o x id e r e d u c ta s e
+
S
N A D P H + H O
|| –

— C H 3
D ic o u m a r o l
— R
||
O
V ita m in K
( q u in o n e fo r m )
 At physiological pH, the two carboxyl groups
attached to the g-carbon of carboxyglutamate
residues are ionized

 The two negatively charged carboxyl groups

act as a Ca++-binding site

OOC COO

CH
H CH2 O
N CH C
 Thus, the function of vitamin K is to create Ca++-
binding sites on the prothrombin molecules as a
result of which they become biologically active

 Similar Ca++-binding sites are probably created

by vitamin K on Factors VII, IX and X also
 Sources

 Green leafy vegetables (alfalfa, spinach,

cabbage etc) are rich in vitamin K

 Cauliflower and peas are also good sources

 Liver is a fair source

 Small amounts are present in milk and eggs

also
 A very important source is bacterial synthesis in
the intestine

 The intestinal bacteria synthesize a good deal of

vitamin K which is available to the host
 Requirement

 The exact requirement is not known as

sufficient quantities are synthesized by intestinal
bacteria

 Vitamin K supplements are required only when

the intestinal bacteria are destroyed or when fat
absorption is impaired

 The normal daily requirement is probably less

than 0.1 mg
 Deficiency

 Deficiency of vitamin K disturbs coagulation of

blood

 Slight injuries can cause prolonged bleeding in

a subject deficient in vitamin K

 Diagnosis can be made by measuring

prothrombin time

 Prothrombin time is prolonged in vitamin K

deficiency
 Prothrombin time may also be prolonged in
patients with liver damage due to inability of the
liver to synthesize prothrombin

 These two conditions can be distinguished by

parenteral administration of vitamin K

 Prothrombin time returns to normal in subjects

deficient in vitamin K but not in patients with
liver damage