BMS 1.1,CC 1.

1D
CHEMISTRY CARBOHYDRATES
 BIOCHEMISTRY

• What is Biochemistry?
• It is a branch of science that deals with the chemical nature and the
chemical behavior of living matter.
• Biochemistry was introduced by a Germany chemist CARL NEUBERG
in the year 1903.
• Biochemistry takes the account on the study related to the nature of
the chemical constituents of the living matter, their transformation in
the biological systems and the energy changes associated with the
transformation.
• Biochemistry has been named as the biology of chemistry or the
chemical biology. It’s also known as the chemical physiology due to
the origin as an offspring from human physiology, when it was
realized that the chemical analysis of the urine, blood and the other
natural fluids that could assist in the diagnosis of particular diseases.
• Physiology covers the study of the normal functioning and the
phenomena of the living things.
• While Biochemistry deals particularly with the chemical aspects of
these functional and phenomenon.
 MODERN BIOCHEMISTRY:
• There are two branches of biochemistry.
• Descriptive Biochemistry
• Dynamic Biochemistry
A. DESCRIPTIVE BIOCHEMISTRY:
• Descriptive Biochemistry is concerned with the qualitative and quantitative
characterization of various cell components.
B. DYNAMIC BIOCHEMISTRY:
• Dynamic Biochemistry deals with elucidation of the nature and the
mechanism of the reactions involving these cell components.
• There are various disciplines that are emerging from biochemistry
including Enzymology, endocrinology, clinical biochemistry, molecular
biochemistry, agricultural biochemistry, pharmaceutical biochemistry etc
• The living things under study in biochemistry are characterized by their
capacity of movements, power of growth, respiratory activity, sense of
irritability and reproduction.
• Some of the major characteristics include;
1. Have highly complicated, organized structures and contain number of
different molecules. E. coli contains 5000 different kinds of organic
compounds like proteins and the nucleic acids.
2. Have specific purposes or function like heart, lungs, brain and the
intracellular structures like nucleus
3. Have the ability to extract nutrients, transform and use energy from
the environment either from organic nutrients or the radiant energy
of the sun
4. Capacity for self replication, these results in the growth and
reproduction in identical forms, mass, shape and internal structure.
• In an oriented research by the biochemist and microbiologist, they use a
general scientific methods approach which is also called HYPOTHETICO-
DEDUCTIVE METHOD.
 Hierarchy of biomolecular organization;
• These are simple molecules that are the Units for Building Complex
Structures like Metabolites and Macromolecules, Organelles like a
membrane. The Unit of Life is the Cell.
Distinctive properties of a living system:
• Organisms are complicated and highly organized
• Biological structures serve functional purposes
• Living systems are actively engaged in energy transformations
• Living systems have a remarkable capacity for self-replication.
 Bio-molecule and biopolymer:
• A biomolecule is any molecule that is produced by a living organism,
including large macromolecules such as proteins, polysaccharides,
lipids, and nucleic acids, as well as small molecules such as primary
metabolites, secondary metabolites, and natural products.
Biopolymer:
• Is a polymeric substance (as a protein or polysaccharide) formed in a
biological system.
• There are three main classes of biopolymers based on the differing
monomeric units used and the structure of the biopolymer formed:
1. Polynucleotides
• Which are long polymers, composed of 13 or more nucleotide
monomers;
2. Polypeptides
• Which are short polymers of amino acids and
3. Polysaccharides
• Which are often linear bonded polymeric carbohydrate structures.
• Biomolecules and biopolymers are the biological molecules that
particitipate in;
• In the physiological functioning of living things.
• They are either organic or inorganic in nature.
• They may be classified as monomeric or polymeric compounds.
• They include;
• Carbohydrates, Lipids, Proteins and amino acids, Enzymes, Plasma
proteins, nucleic acids, Immunoglobulin, Haemoglobin, Vitamins,
Prostaglandins.
• Biomolecules are hydrocarbon in nature, made up of elements H, O, C
and N. Make up 99+% of atoms in the human body. They are biologically
important molecules due to their ability to form covalent
 bonds by electron-pair sharing.
ELEMENT PERCENTAGE
1. Oxygen 63
2. Hydrogen 25.2
3. Carbon 9.5
4. Nitrogen 1.4
• The bond energies of covalent bonds
 Bond Energy kJ/mol
1. H-H 436
2. C-H 414
3. C-C 343
4. C-O 35
PROPERTIES OF BIOMOLECULES REFLECT THEIR FITNESS TO THE
LIVING CONDITION
• Macromolecules and Their Building Blocks Have a “Sense” or
Directionality,
• Macromolecules are Informational,
• Biomolecules Have Characteristic Three-Dimensional Architecture,
• Weak Forces Maintain Biological Structure and Determine
Biomolecular Interactions,
• Like van der Waals: 0.4-4.0 kJ/mole,
• Hydrogen bonds: 12-30 kJ/mole, Ionic bonds: 20 kJ/mole,
Hydrophobic interactions: <40 kJ/mole.
 CARBOH YDRATES:

• They are polyhydroxy aldehydes or ketones and their derivatives or

compounds which yield these on hydrolysis.
• Carbohydrates are hydrates of carbon.
• The carbohydrates are termed as “sugars” and are the most abundant
biomolecules in nature.
• They are referred to as saccharides (`sakcharone’=sugars/sweetness).
• They have a small molecular formula and have a sweet taste.
Importance of Carbohydrates:
• Polymeric carbohydrates–above all starch, as well as some
disaccharides–are important (but not essential) components of
food.
1. In the gut, they are broken down into monosaccharides and
reabsorbed in this form.
The form in which carbohydrates are distributed by the blood of
vertebrates is glucose (blood sugar).
2. This is taken up by the cells and either broken down to obtain energy
(glycolysis) or converted into other metabolites.
3. Several organs (particularly the liver and muscles) store glycogen as a
polymeric reserve carbohydrate.
4. The glycogen molecules are covalently bound to a protein,
glycogenin. Polysaccharides are used by many organisms as building
materials.
• For example, the cell walls of bacteria contain murein as a stabilizing
 component while in plants cellulose and other polysaccharides fulfill
this role.
5. Oligomeric or polymeric carbohydrates are often covalently bound to
lipids or proteins. The glycolipids and glycoprotein formed in this
way are found in cell membranes (center). Glycoproteins also occur
in the blood in solute form (plasma proteins) and, as components of
proteoglycans, form important constituents of the intercellular
substance.
CLASSIFICATION OF
CARBOHYDRATES:
• They are classified into three;
1. Monosaccharides
2. Oligosaccharides
3. Polysaccharides.
MONOSACCHARIDES
• Also called simple sugars.
• Are those sugars which cannot be hydrolyzed into simpler forms.
• They can be subdivided further;
a. Classification according to the number of carbon atoms;
• Trioses, Tetroses, Pentoses, Hexoses, Heptoses.
b. Classification according to functional group:
1. Aldoses
• Glycerol, Erythrose, Ribose, Glucose, Glucoheptose.
2. Ketoses
• Dihydroxyacetone, Erythrulose, Ribulose, Fructose, Sodoheptose.
No. of Carbon Atoms Name Compounds Aldoses Ketoses

3 Trioses Glycerol Glycerol Dihydroxyacetone

4 Tetroses Erythroses Erythroses Erythruloses

5 Pentoses Riboses Riboses Ribuloses

6 Hexoses Glucoses Glucoses Fructoses

7 Heptoses Glucoheptoses Glucoheptoses Sodoheptoses

OLIGOSACCHARIDES:
Oligo means ‘few’.
• These are sugar compound that yield two to ten molecules of either
same or different monosaccharides on hydrolysis.
• Based on the number of monosaccharides present they are sub-divided
as:
DISACCHARDIDES:
• These yield 2 molecules of monosaccharides on hydrolysis.
Example;
• Sucrose, lactose, maltose, cellobiose, trehalose, gentiobiose, melibiose.
TRISACCHARDES:
• These yield 3 molecules of monosaccharides on hydrolysis.
 Example;
• Rhaminose, gentianose, raffinose, melecitose.
TETRASACCHARIDES:
• These yield 4 molecules of monosaccharides on hydrolysis.
Example;
• Starchyose, scorodose
PENTASACCHARIDES:
• These yield 5 molecules of monosaccharides on hydrolysis.
Example; Verbascose.
FORMULATION OF
CARBOHYDRATES:
1. ASSYMETRIC CARBON ATOM:
• Asymmetric carbon atom is the carbon which is attached to 4
different groups or atoms.
• Any substances containing one or more asymmetric carbon atom
shows 2 properties;
• Optical activity and optical isomerism
i. OPTICAL ACTIVITY :
•  Is the ability of a substance to rotate plane polarized light either to
the right or to the left.
1. If the substance rotates plane polarized light to the right, it is
called dextrorotatory (d) or ( + ) .
2. If it rotates it to the left, it is called levorotatory (l) or ( - ).
• Glucose contains 4 asymmetric carbon atoms. It is dextrorotatory, so
it is sometimes named dextrose.
• Fructose contains 3 asymmetric carbon atoms. It is levorotatory so it
is sometimes called laevulose.
SPECIFIC ROTATION:
• It is the angle of rotation specific for each optically active substance
 when the concentration of substance is 100 g / dl and the length
of measuring tube is 10 cm.
E.g specific rotation for glucose is ( +52.5°)and for fructose is ( -91°).
RACEMIC MIXTURE:
• It is the mixture containing equal number of molecule of 2 optically active
sugars. (When equal amounts of dextrorotatory and laevorotatory isomers
are present, the resulting mixture has no optical activity, since the activities
of each isomer cancel each other).
• One is dextrorotatory and the other is levorotatory. Thus, it shows no
optical activity (Provided that the angle of rotation is equal in both sides).
 Resolution:
• It is the separation of optically active isomers from a racemic mixture.
Isomerism:
• Iso refers to Equal and Meros refers to Part.
• It was applied by Jones Jacob in1827.
• Isomerism is a phenomenon in which certain compounds posses
same molecular formula but exist in different forms owing to the
different organisation of atoms and the phenomenon is referred to as
Isomerism.
 CLASSIFICATION OF ISOMERS:

ISOMER
Structural Stereo Isomer.
Chain Position Function gp. -Geometrical or Cis- trans Isomer.
Isomer. Isomer.
Structural isomer:
• These are compounds that have the same molecular formula but differ in
the structures.
i. Chain isomer: difference in the length of the carbon chains.
ii. Position isomers: Differ in the position of substituent groups.
iii. Functional group: Posses different functional groups.
Stereoisomer:
• These are compounds that have the same molecular formula but
differ only in the spatial configuration.
• While writing the molecular formula of monosaccharides, the spatial
arrangements of H and OH groups are important, since they contain
asymmetric carbon atoms.
 The reference molecule is glyceraldehyde (glycerose) which has a
single asymmetric carbon atom.
Geometrical isomer:
• Example maleic acids (cis – form) and fumaric acids (trans- form).
• The number of possible stereoisomers depends on the number of
asymmetric carbon atoms by the formula 2n. where n is the number of
asymmetric carbon atoms.
Optical isomers:
• Differ in the deposition of various atoms or groups of atoms in space around
the asymmetric carbon atoms.
• i.e D and L form of Glucose.
OPTICAL ISOMERISM:
•  It is the ability of a substance to present in more than one form
(isomer).
• A substance containing one asymmetric carbon atom can exist in a
number of isomers = 2n,
• Where n is the number of asymmetric carbon atoms.
e. g. glucose has 4 asymmetric carbon atoms, so the number of its
isomers equal 24 = 2 x 2 x 2 x 2 = 16 isomers.
 Configuration ( Enantiomers ):
• The simplest carbohydrates are monosaccharides trioses ;
for example glyceraldehyde which has one asymmetric carbon and
thus 2 optically active forms.
• L and its mirror image D forms . 
Reference sugar:
• It is glyceraldehydes which may be present in (D)form in which –OH
group attached to asymmetric carbon atom is on the right side and
(L ) form in which the same –OH group is on the left side .
1). All other monosaccharides are considered to be derived from
reference sugar glyceraldehyde. They are classified into D and L
forms according to the position of –OH attached to the carbon atom
adjacent to last -CH2OH e. g. carbon atom number 5 in glucose.
2). Most of the monosaccharides occurring in mammals are of D
configuration (form). However, a sugar may be dextrorotatory (d)
or levorotatory (l) irrespective of its D or L forms.
Anomeric carbon and anomers:
 Anomeric carbon:
• Is the asymmetric carbon atom obtained from active carbonyl sugar
group.
• Carbon number 1 in aldoses and carbon number 2 in ketoses.
Anomers :
• These are isomers obtained from the change of position of hydroxyl
attached to the anomeric carbon.
E. g .α and β glucose are 2 anomers. Also α and β fructose are 2
anomers.
 MUTAROTATION
• This is the gradual specific change in any optically active substance
having free aldehyde (-CHO) or ketone (C=O) group.
• When an aldohexose is first dissolved in water and the solution is put in
optical path so that plane polarized light is passed, the initial optical
rotation shown by the sugar gradually changes until a constant fixed
rotation characteristic of the sugar is reached.
• α -Glucose freshly dissolved in water has specific rotation of +112˚. β -
Glucose when freshly dissolved in water, has specific rotation of +19˚.
When both anomers are left for some time, α and β sugars slowly change
into an equilibrium mixture which has specific rotation of +52.5˚.
• This is explained by the fact that D-glucose has two anomers, alpha
and beta varieties. These anomers are produced by the spatial
configuration with reference to the first carbon atom in aldoses and
second carbon atom in ketoses.
• Hence, these carbon atoms are known as anomeric carbon atoms.
Thus α-D-glucose has specific rotation of +112° and β-D-glucose has
+19°. Both undergo mutarotation and at equilibrium one-third
molecules are alpha type and 2/3rd are beta variety to get the specific
rotation of +52.5°. The differences between α and β anomeric forms
are dependent on the 1st carbon atom only.
 CARBOHYDRATES
FORMULATION.
• The formulation of carbohydrates was done by Fischer and Haworth.
• The open chain projection formula and hemi acetal ring structure of
glucose were proposed by Emil Fischer in 1883, and hence called
Fischer's formula.
• Later it was shown that the glucose exists in biological systems, not as a
rectangle, but as a pyranose ring by Sir Walter Haworth in 1925.
• In Haworth projection during the year 1929, carbohydrates are
formulated in a cyclic structure as:
1. Pyranosyl-a 6 member ring.
2. Furanosyl- a 5 member ring.
• The conversion of Fischer’s structure to Haworth structure occurs after
the condensation reaction has taken place at the C1 of Aldoses and C2 of
the Ketoses hence the O-Glycosidic linkage in the C1-C5 in aldose sugars
and C2-C5 in ketose sugars.
DISACCHARIDES:
• Among the oligosaccharides, disaccharides are the most common.
• Consists of two monosaccharide units (similar or dissimilar) held together
by a glycosidic bond.
• They are crystalline, water-soluble and sweet to taste.
• They are of two types:
1. Reducing disaccharides with free aldehyde or keto group
e.g. maltose, lactose.
2. Non-reducing disaccharides with no free aldehyde or keto group
e.g. sucrose, trehalose.
A. LACTOSE:
• Also called as milk sugar.
• Found in cow milk, mother’s milk.
• Made up of β D galactose and β D glucose sugars held together by β (1-
4) glycosidic linkages.
 Properties:
• Crystalline solid with a melting point of 203 ̊ C,
• Dextrorotatory with a specific rotation of +52.5 ̊,
• Less soluble in water, and less sweet than sucrose.
• Experiences mutarotation and it is a reducing monosaccharide (reduces
Cu²+ to cu+) and can form osazone and oxime.
• It undergoes hydrolysis or digestion in the presence of lactase enzyme to
form galactose and glucose molecules.
• In the absence of lactase enzyme it leads to lactose intolerance in adults.
B: SUCROSE:
• Also called table sugar /cane sugar/beet sugar.
• It is found in beet, cane, and maple. It is commonly found in the
photosynthetic plants.
• It is made up of glucose and fructose held together by α1-2 glycosidic
linkage.
• It is a non-reducing sugar.
Properties:
• It is crystalline solid in nature,
• Soluble in water,
• Has a melting point of 180 ̊C and forms brown substances (caramel)
and forms chars in presence of sulphuric acid.
• Dextrorotatory with specific rotation of +66.7 ̊ C.
• Hydrolysis in the presence of sucrase enzyme results in the formation
of glucose (dextrorotatory +52.5 ̊ C) and fructose (laevorotatory-92.0
̊C).
C. MALTOSE
• Also called malt sugar.
• Occurs in starch, malt.
• Sprouting cereals have large amount of amylase enzymes.
• Maltose contains free hemiacetal group hence a reducing sugar,
• It is crystalline in nature with a melting point of 160-165 ̊ C,
• Soluble in water and is dextrorotatory due to the free aldehyde group.
It forms an osazone in the presence of phenylhydrazine.
• Exhibit mutarotation.
• On hydrolysis it yields two identical glucose molecules in dilute acids
or maltase enzyme in the intestines.
 ISOMALTOSE:
• It is similar to the maltose sugar except it contains Alpha 1-6
glycosidic linkage.
• On hydrolysis in the presence of isomaltase it yields glucose
molecules.
Cellobiose:
• Released during the digestion of cellulose polysaccharides in the
presence of cellulase enzyme.
• Cellobiose is identical to maltose. Has alpha 1-4 glycosidic linkage.
• It is crystalline in nature with a melting point of 225 ̊C.
• Soluble in water and is dextrorotatory due to the hemiacetal group.
• Undergoes mutarotation and is reducing sugar.
• It forms osazone in the presence of phenylhydrazine.
 POLYSACCHARIDES- Greek- Many.
• Also referred to as polymeric anhydride of simple sugars.
• Consist of repeat units of monosaccharides or their derivatives.
• Held together by glycosidic bonds.
• They are primarily concerned with two important functions.
i). Structural and
ii). Storage of energy
• Polysaccharides are linear as well as branched polymers.
• They are classified into two;
1. HOMOPOLYSACCHARIDES
2. HETEROPOLYSACCHARIDES.
• These are compounds of sugars that yield more than 10
monosaccharides on hydrolysis.
• They are further classified depending on the monosaccharides that
are yield on hydrolysis (same or different).
1. Homopolysaccharides:
• Yield same monosaccharides on hydrolysis.
Example-
• Starch, glycogen, inulin, cellulose, pectin, and chitin.
2. Heteropolysaccharides:
• Yield different monosaccharides on hydrolysis.
Example;
• Hyaluronic acids, chondroitin sulphate, specific soluble sugars of
Pneumonococcus type III.
 HOMOPOLYSACCHARIDES
1. Starch:
• Starch occurs or is found in legumes, cereals, potatoes, and other
vegetables.
• It is the most important reserved food materials.
• Also it is the most commonly ingested carbohydrates by humans.
• Natural starches consist of two components;
i. Amylose: 15 -20% (long unbranched and straight chained) and
ii. Amylopectin: 80-85% (branched chain polysaccharide).
• When treated with boiling water, 10-20% is solubilized; this part is
called amylose.
• Amylose is made up of glucose units with alpha-1,4 glycosidic
linkages to form an unbranched long chain with a molecular weight
400,000 D or more.
• The insoluble part absorbs water and forms paste like gel; this is
called amylopectin.
• Amylopectin is also made up of glucose units, but is highly branched
with molecular weight more than 1 million. The branching points are
made by alpha-1,6 linkage (similar to isomaltose).
 Properties:
• Starch is a white soft amorphous powder and lacks sweetness,
• Insoluble in water, alcohol,
• It experiences mutarotation not specific rotation of +196ᵒC.
Hydrolysis:
• It undergoes acid hydrolysis in the presence of amylase enzyme to
yeild D-glucose molecules.
• And in addition of iodine solution it turns from blue to colourless on
complete hydrolysis.
i. Starch will form a blue colored complex with iodine; this color
disappears on heating and reappears when cooled. This is a sensitive
test for starch.
• It is non reducing because the free sugar groups are negligible in
number.
ii. When starch is hydrolysed by mild acid, smaller and smaller
fragments are produced.
iii. Thus hydrolysis for a short time produces amylodextrin which gives
violet color with iodine and is nonreducing. Further hydrolysis
produces erythrodextrin which gives red color with iodine and mild
reduction of Benedict's solution. Later achrodextrins (no color with
iodine, but reducing) and further on, maltose (no color with iodine,
but powerfully reducing) are formed on continued hydrolysis.
Starch Iodine solution colour change(blue)

Soluble starch Blue

Amylodextrin Purple

Erythrodextrin Red

Achroodextrin Colourless

Maltose Colourless

Glucose Colourless
 Amylase Enzyme:
• This is the key enzyme that participates in the hydrolysis of the starch
molecules:
i. α-amylase : found in the saliva and pancreatic juice.
ii. β-Amylase : found in the sprout grains and malts.
i. Salivary amylase and pancreatic amylase:
• They are alpha-amylases, which act at random on alpha1,4 glycosidic
bonds to split starch into smaller units (dextrins), and finally to alpha-
maltose.
ii. Beta-amylases are of plant origin (almond, germinating seeds, etc)
which split starch to form beta-maltose. They act on amylose to split
maltose units consecutively. Thus the enzyme starts its action from one
end.
iii. When beta-amylase acts on amylopectin, maltose units are liberated
from the ends of the branches of amylopectin, until the action of
enzyme is blocked at the 1,6-glycosidic linkage. The action of beta-
amylase stops at branching points, leaving a large molecule, called limit
dextrin or residual dextrin.
Differences between amylose and amylopectin:
Amylose Amylopectin

Low molecular weight of about 60,000 High molecular weight of about 500,000

Occur to an extend of about 15%-20% Occur to an extend of about 80%-85%

Gives a blue colour with iodine solutioin Gives a reddish –violet colour with iodine solution

Structure is unbranched or linear Structure: branched

Posses aC1-C4 glycosidic linkages Posses C1-C4 and C1-C6 glycosidic linkages
 GLYCOGEN:
• Major reserve food in animals and stored in liver and muscles.
• It can be branched or unbranched and resembles amylopectin.
• It is composed of glucose units joined by alpha-1,4 links in the straight
chains. It also has alpha-1,6 glycosidic linkages at the branching
points.
• Molecular weight of glycogen is about 5 million. Innermost core of
glycogen contains a primer protein, Glycogenin.
• Glycogen is more branched and more compact than amylopectin.
 Properties:
• Powder and more soluble in water.
• Undergoes precipitation in aqueous solution i.e ethyl alcohol, non-
reducing sugar, react with iodine to form red colour.
Hydrolysis:
• Hydrolysis is done by the enzyme α 1-4 glucanmaltohydrolase to yield
maltose and on acid hydrolysis it yields glucose.
 INULIN:
• It is a long chain homoglycan composed of D-fructose units with
repeating beta-1,2 linkages.
• It is the reserve carbohydrate present in various bulbs and tubers such
as chicory, dahlia, dandelion, onion, garlic.
• It is clinically used to find renal clearance value and glomerular
filtration rate. (Used for assessing kidney function through
measurement of glomerular filtration rate (GFR)).
• Inulin is made up of 30-35 monomer units of fructose held together by
β 1-2 fructosidic linkage
• On hydrolysis it yields fructose molecules.
Properties:
• It is a tasteless powder and is insoluble in cold water,
• Easily soluble in warm water to form a colloidal solution which does
not form gel on cooling.
• Its non-reducing sugar, no colour is formed in the presence of iodine.
 CELLULOSE:
• Structural polysaccharides, occurs exclusively in plants and it is the
most abundant organic substance in plant kingdom.
• A predominant constituent of plant cell wall. Cellulose is totally
absent in animal body.
• Molecular weight of 200,000 to 2,000,000 Da with 1250-12,500
glucose residues.
• Has β D glucose molecules with β 1-4 gycosidic linkage.
 Properties:
• Fibrous, tough, white solid, insoluble in water but soluble in amonical
cupric solution (Schweitzer’s reagent).
• Forms no colour with iodine reagent and lacks sweetness.
Hydrolysis:
• It undergoes hydrolysis in the presence of an acid HCl or H2SO4 or
concentrated base like NaOH in the presence of cellulase enzyme.
 PECTIN
• It occurs in the tissues of young plants especially in ripe fruits i.e
guavas, apples and pears.
• A major component in the cell wall and adjacent walls.
• Made up of α –D galacturonic acids with a high molecular weight.
• With C1-C4 galacturonic linkage.
CHITIN:
• lt is a structural polysaccharide found in the exoskeleton of some
invertebrates e.g. insects, crustaceans.
• Composed of N-acetyl D-glucosamine units held together by β(1→ 4)
glycosidic bonds.
Hemicellulose
• Composed of a mixture of monosaccharides.
• The most common monosaccharides comprising the backbone of
hemicelluloses are:
Xylose, mannose, and galactose.
• Baking soda maintains vegetable color but breaks down their
hemicelluloses (making them mushy).
• The common side chains are:
Arabinose, glucuronic acid, and galactose.
HETEROPOLYSACCHARIDES: HETEROGLYCANS OR
MUCOPOLYSACCHARIDES
• They are of two types:
i. Neutral mucopolysaccharides and
ii. Acidic mucopolysaccharides.
I. Neutral mucopolysaccharides.
a. Pneumococci polysaccharides.
b. Blood group substances.
c. Nitrogenous neutral substances.
II. Acidic mucopolysaccharides.
• They are of two types:
(i). Sulphate free and
(ii). Sulphate containing.
i. Sulphate free
a. Chondroitin.
b. Hyaluronic acids.
ii. SULPHATE CONTAINING
a. Chondrotin sulphate.
b. Heparin.
c. Keratin sulphate.
 Neutral mucopolysaccharides:
a. Pneumococci polysaccharide:
• Found in the pneumococci capsules they contain the glucosamine and
glucuronic acids.
b. Blood group substances:
• Contain both peptide and carbohydrates.
• The monosaccharides include Galactose, fucose, N acetylated
galactosamine and N acetylated glucosamine.
c. Nitrogenous neutral mucopolysaccharides:
• Firmly bound to proteins ie ovalbumin which contains the mannose and
glucosamine.
Sulphate free acidic mucopolysaccharides:
a. Hyaluronic acids:
• It is found in the synovial fluids, eye, and umbilical cord.
Importance:
• It is part of the synovial fluid that makes it a good lubricant in the joints.
Hyaluronic acids are made up of D-glucuronic acid and N-Acetyl-D-
glucosamine (NAG) alternating in the chain held together by β-1-3 and
β1-4 linkage.
Hydrolysis:
• It yields equimolar mixture of of D glucuronic acids, D-glucosamine, and
acetic acids in the presence of Hyaluronidase enzyme mainly present in
the sperm cells this facilitate the fertilization of the ovum.
• Hyaluronidase is an enzyme that breaks (β 1 → 4 linkages) hyaluronic acid
and other CAC. This enzyme is present in high concentration in testes,
seminal fluid, and in certain snake and insect venoms. Hyaluronidase of
semen is assigned an important role in fertilization as this enzyme clears
the gel (hyaluronic acid) around the ovum allowing a better penetration
of sperm into the ovum. Hyaluronidase of bacteria helps their invasion
into the animal tissues
b. Chondroitin:
• Found in the cornea.
• Has been isolated from cranial cartilages.
• It is made up N-Acetyl Galactosamine and D-Glucuronic acids.
Hydrolysis
• It yields equimolar solution of D glucuronic acids, D-galactosamine,
and acetic acids held together by β-1-4 linkages.
• They are principal mucopolysaccharide in the ground substance of
mammalian tissues and cartilage.
• Occur in combination with proteins and are called as
Chondroproteins.
• Four chondroitin sulphates have been isolated so far.
• They are named as chondroitin SO₄ A, B, C and D.
3. Sulphate containing acid mucopolysaccharides:
1. Chondroitin sulphates:
• It is found in the mammalian tissues and cartilage.
• Also present in ground substance of connective tissues widely
distributed in cartilage, bone, tendons, cornea and skin.
• Composed of repeating units of glucuronic acid → beta-1,3-N-acetyl
galactosamine sulphate → beta-1, 4 and so on
• There are four classes of chondroitin sulphates A, B, Cand D. The
chondroitin sulphate A and B are the most common.
i. Chondroitin sulphate A:
• Found in the cartilages, adult bone and cornea.
• Made up of repeated units of N-Acetyl –D-Galactosamine and
esterified N-Acetyl galactosamine at C4. N-Acetyl galactosamine is
esterified with SO₄ in position 4 of galactosamine.
ii. Chondroitin sulphate B:
• Found in the skin, cardiac valves and tendons.
• Also isolated from aortic wall and lung.
Structure
• It is made up of repeating L-iduronic acids, N-Acetyl –D-
galactosamine sulphate. Sulphate moiety is present at C₄ of N-acetyl
galactosamine molecule.
iii. Chondroitin sulphate C:
• Found in the tendons and cartillages.
• Made up of N-Acetyl –D-Galactosamine and esterified N-Acetyl
galactosamine. Structure of chondroitin SO₄ C is the same as that of
chondroitin SO₄ A except that the SO₄ group is at position 6 of
galactosamine molecule instead of position 4.
iv. Chondroitin sulphate D:
• It has been isolated from the cartilage of shark.
• Similar to chondroitin sulphate C but the difference is sulphate is attached
to C 2or 3 of L-Uronic acids. (It resembles in structure to chondroitin SO4
C except that it has a second SO4 attached probably at carbon 2 or 3 of
uronic acid moiety).
B. Heparin:
• It is an anticoagulant present in liver.
• Found in the mast cells in the liver, thymus, spleen, walls of large arteries,
skin and in small quantities in blood.
 Structure.
• Made up of repeating disaccharide units of D-Glucosamine and either
of the two uronic acids (D-glucuronic acid and L-Iduronic acid).
• The -NH₂ group at C2 and OH group at C6 of D-Glucosamine (Glc N)
are sulphated. A few may contain acetyl group on C2 of D-
Glucosamine. In addition, the OH group of C2 of uronic acids, D-
Glucuronic acid and/or L-Iduronic acid, are sulphated.
• Heparin is anticoagulant that inhibits the binding (clotting) of the
antithrombin III.
• It is also used in vivo in suspected thrombo-embolic conditions to
prevent intravascular coagulation. It activates antithrombin III, which in
turn inactivates thrombin, factor X and factor IX.
Properties:
• It is strongly acidic due to sulphuric acid groups and readily forms salts.
C. Keratan sulphate:
• Found in the cartillage and cornea.
• Made up of repeated disaccharide units of N-Acetyl glucosamine and
galactose. Sulphate present at C1-C6 of both N-Acetyl glucosamine
 and galactose.
• They are two types Keratan sulphate I and keratan sulphate II and
both are found in the tissues.
Types:
• Two-types have been described.
• They are found in tissues combined with proteins.
a. Keratan SO₄ I:
• It occurs in cornea.
• Linkage is between N-acetyl glucosamine and Asparagine residue to
form the N-glycosidic bonding.
b. Keratan SO₄ II:
• It occurs in skeletal tissues.
• The linkage to protein is by way of -OH groups on serine and
threonine residues of the protein.
 PROTEOGLYCANS
• Are high molecular weight, complex molecules with diverse structures
and functions.
• They are polyanionic substances containing a core protein to which at
least one glycosaminoglycan (mucopolysaccharide) chain is covalently
attached.
• They are major components of connective tissue and participate with
other structural protein constituents, namely, collagen and elastin, in
the organization of the extracellular matrix.
• Core proteins are covalently linked to glycosaminoglycans (GAGs).
• Any of the Glycosaminoglycans viz.
Hyaluronic acid (HA); keratan sulphates I and II, chondroitin
sulphates A, B, C, heparin and heparan sulphate can take part in its
formation.
• The amount of carbohydrates in proteoglycans is much greater (up to
95%) as compared to glycoproteins.
Linkages:
• Three types of linkages between GAG and coreprotein is observed.
i. O-glycosidic linkage:
• Formed between N-acetyl galactosamine (GalNAc) and serine or
threonine of the core protein.
Example:
• Typically seen in keratan SO₄ II.
ii. N-glycosylamine linkage:
• Formed between N-acetyl glucosamine (GlcNAc) and amide N of
asparagine (ASn) of core protein.
Example:
• Typically seen in keratan SO₄ I and N-linked glycoproteins.
iii. O-glycosidic linkage:
• Formed between xylose (Xyl) and serine of the protein.
• This bond is unique to proteoglycans.
Functions of Proteoglycans
1. As a constituent of extracellular matrix or ground substance:
• Interacts with collagen and elastin
2. Acts as polyanions:
• GAGS present in proteoglycans are polyanions and hence bind to
 polycations and cations such as Na and K. Thus attracts water by
osmotic pressure into extracellular matrix contributing to its turgor.
3. Acts as a barrier in tissue:
• Hyaluronic acid in tissues acts as a cementing substance and contributes
to tissue barrier which permit metabolites to pass through but resist
penetration by bacteria and other infective agents.
4. Acts as lubricant in joints:
• Hyaluronic acid in joints acts as a lubricant and shock absorbant. Intra
articular injection of hyaluronic acid in knee joints is used to alleviate pain
in chronic osteoarthritis of knee joints.
5. Role in release of hormone:
• Proteoglycans like hyaluronic acid are present in storage or secretory
granules, where they play part in release of the contents of the
granules.
6. Role in cell migration in embryonic tissues:
• Hyaluronic acid is present in high concentration in embryonic tissues
and is considered to play an important role in cell migration during
morphogenesis and wound repair.
7. Role in glomerular filtration:
• Proteoglycans like hyaluronic acid is present in basement membrane
(BM) of glomerulus of kidney where it plays important role in charge
selectiveness of glomerular filtration.
8. Role as anticoagulant in vitro and in vivo:
– In vitro, heparin is used as an anticoagulant. 2 mg/10 ml of blood is
used. Most satisfactory anticoagulant as it does not produce a change
in red cell volume or interfere with its subsequent determinations.
– In vivo, heparin is an important anticoagulant. It binds with factor IX
and XI , but its most important action is with plasma antithrombin III .
Binding of heparin to lysine residues in antithrombin III produces
 conformational change which promotes the binding of the latter to
serine protease thrombin which is inhibited, thus fibrinogen is not
converted to fibrin.
• Four naturally occurring thrombin inhibitors in plasma are;
a. Antithrombin III (75% of the activity).
b. α₂ –macroglobulin contributes remainder.
c. Heparin cofactor II.
d. α₁– antitrypsin.
9. Role as a coenzyme:
• Heparin acts in the body to increase the activity of the enzyme
Lipoprotein lipase . Heparin binds specifically to the enzyme present
in capillary walls, causing a release of the enzyme into the circulation.
Hence heparin is called as Clearing factor.
10. As a receptor of cell:
• Proteoglycans like heparan sulphate are components of plasma
membrane of cells, where they may act as receptors and can
participate in cell adhesion and cell-cell interactions.
11. Role in compressibility of cartilages:
• Chondroitin sulphates and hyaluronic acid are present in high
concentration in cartilages and have a role in compressibility of
cartilage in weight bearing.
12. Role in sclera of eye:
• Dermatan sulphate is present in sclera of the eye where it has an
important function in maintaining overall shape of the eye.
13. Role in corneal transparency:
• Keratan sulphate I is present in cornea of the eye and lies between
 the collagen fibrils. It plays an important role in maintaining corneal
transparency.
• EXTENDED
Glycosaminoglycans
(mucopolysaccharides)
• Are complex carbohydrates containing amino sugars and uronic acids.
They may be attached to a protein molecule to form a proteoglycan.
• Proteoglycans provide the ground or packing substance of connective
tissue. They hold large quantities of water and occupy space, thus
cushioning or lubricating other structures, because of the large
number of ´OH groups and negative charges on the molecule, which,
by repulsion, keep the carbohydrate chains apart.
• Examples are hyaluronic acid, chondroitin sulfate, and heparin.
Glycoconjugates: Proteoglycans,
Glycoproteins, and Glycolipids
• In addition to their important roles as stored fuels (starch, glycogen,
dextran) and as structural materials (cellulose, chitin, peptidoglycans),
polysaccharides and oligosaccharides are information carriers, they
serve as destination labels for some proteins and as mediators of
specific cell-cell interactions and interactions between cells and the
extracellular matrix.
• Specific carbohydrate containing molecules act in cell-cell recognition
and adhesion, cell migration during development, blood clotting, the
immune response, and wound healing, to name but a few of their
many roles.
• In most of these cases, the informational carbohydrate is covalently
joined to a protein or a lipid to form a glycoconjugate, which is the
biologically active molecule.
Proteoglycans
• Are proteins that contain covalently linked glycosaminoglycans(GAGs).
• At least 30 have been characterized and given names such as syndecan,
betaglycan, serglycin, perlecan, aggrecan, versican, decorin, biglycan,
and fibromodulin.
• The proteins bound covalently to glycosaminoglycans are called core
proteins.
• They (Proteoglycans)vary;
a. In tissue distribution,
b. Nature of the core protein,
c. Attached glycosaminoglycans, and
d. Their function.
• They have proved difficult to isolate and characterize, but the use of
recombinant DNA technology (DNA molecules formed by laboratory
methods of genetic recombination) is beginning to yield important
information about their structures.
• The amount of carbohydrate in a proteoglycan is usually much greater
than that found in a glycoprotein and may comprise up to 95% of its
weight.
• The link proteins interact noncovalently with core protein molecules
from which chains of other GAGs (keratan sulfate and chondroitin
sulfate in this case) project.
• There are at least seven GAGs:
• Hyaluronic acid (hyaluronan), chondroitin sulfate, keratan sulfates I
and II, heparin, heparan sulfate, and dermatan sulfate.
• GAGs are unbranched polysaccharides made up of repeating
disaccharides, one component of which is always an amino sugar
(hence, the name GAG), either D-glucosamine or D-galactosamine.
• The other component of the repeating disaccharide (except in the
case of keratan sulfate) is a uronic acid, either L-glucuronic acid
(GlcUA) or its 5′-epimer, L-iduronic acid (IdUA).
• Proteoglycans are found in every tissue of the body, mainly in the
extra cellular matrix or ground substance.
• There they are associated with each other and also with the other
 major structural components of the matrix, collagen and elastin, in
specific ways.
• Some proteoglycans bind to collagen and others to elastin. These
interactions are important in determining the structural organization
of the matrix. Some proteoglycans (eg, decorin) can also bind growth
factors such as TGF-β, modulating their effects on cells. In addition,
some of them interact with certain adhesive proteins such as
fibronectin and laminin, also found in the matrix.
• The GAGs present in the proteoglycans are polyanions and hence
 bind polycations and cations such as Na+ and K+. This ability attracts
water by osmotic pressure into the extracellular matrix and
contributes to its turgor.
• GAGs also gel at relatively low concentrations because of the long
extended nature of the polysaccharide chains of GAGs and their
ability to gel, the proteoglycans can act as sieves, restricting the
passage of large macromolecules into the extra cellular matrix but
allowing relatively free diffusion of small molecules.
• Because of their extended structures and the huge macromolecular
 aggregates they often form and occupy a large volume of the matrix
relative to proteins.
• GAGs differ from each other in a number of the following properties:
Amino sugar composition,
Uronic acid composition,
Linkages between these components,
Chain length of the disaccharides,
The presence or absence of sulfate groups and their positions of
attachment to the constituent sugars,
The nature of the core proteins to which they are attached,
The nature of the linkage to core protein,
Their tissue and subcellular distribution, and their biologic functions.
Linkages:
• Three types of linkages between GAG and coreprotein is observed.
a. O-glycosidic linkage:
• Formed between N-acetyl galactosamine (GalNAc) and serine or
threonine of the core protein.
 Example: Typically seen in keratan SO₄ II.
b. N-glycosylamine linkage:
• Formed between N-acetyl glucosamine (GlcNAc) and amide N of
asparagine (ASn) of core protein.
Example:
• Typically seen in keratan SO4I and N-linked glycoproteins.
c. O-glycosidic linkage: Formed between xylose (Xyl) and serine of the
protein. This bond is unique to proteoglycans.
 The major features of seven GAGs are summarized below
i). Hyaluronic Acid
• Consists of an unbranched chain of repeating disaccharide units
containing GlcUA and GlcNAc.
• It is present in bacteria and is found in the Extra Cellular Matrix of
nearly all animal tissues, but is especially high in concentration in
highly hydrated types such as skin and umbilical cord, and in bone,
 cartilage, joints (synovial fluid) and in vitreous humor in the eye, as
well as in embryonic tissues.
• It is thought to play an important role in permitting cell migration
during morphogenesis and wound repair.
• Its ability to attract water into the ECM triggers loosening of the
matrix, aiding this process.
• The high concentrations of hyaluronic acid together with chondroitin
sulfates present in cartilage contribute to its compressibility.
ii). Chondroitin Sulfates (Chondroitin 4-Sulfate & Chondroitin 6-
Sulfate)
• Proteoglycans linked to chondroitin sulfate by the Xyl-Ser O-glycosidic
bond are prominent components of cartilage.
• The repeating disaccharide is similar to that found in hyaluronic acid,
containing GlcUA but with GalNAc replacing GlcNAc. The GalNAc is
substituted with sulfate at either its 4′ or its 6′ position, with
approximately one sulfate being present per disaccharide unit.
• Chondroitin sulfates have an important role in maintaining the
structure of the ECM.
• Located at sites of calcification in endochondral bone and are a major
component of cartilage.
• Found in high amounts in the ECM of the central nervous system and,
in addition to their structural function, are thought to act as signaling
molecules in the prevention of the repair of nerve endings after
injury.
 Keratan Sulfates I & II
• They consist of repeating Gal-GlcNAc disaccharide units containing
sulfate attached to the 6′ position of GlcNAc or occasionally of
Galactose.
Keratan sulfate I
• Originally isolated from the cornea.
Keratan sulfate II
• Came from cartilage.
• However, the two GAGs differ in the structural links to the core
 proteins, because the distribution of the two types is not tissue
specific the classification is based on the different protein linkage.
• In the eye, they lie between collagen fibrils and play a critical role in
corneal transparency. Changes in proteoglycan composition found in
corneal scars disappear when the cornea heals.
Heparin
• The repeating disaccharide heparin contains glucosamine (GlcN) and
either of the two uronic acids.
• Most of the amino groups of the GlcN residues are N-sulfated, but a
few are acetylated. The GlcN also carries a sulfate attached to carbon
6.
• The protein molecule of the heparin proteoglycan is unique,
consisting exclusively of serine and glycine residues.
• Approximately two thirds of the serine residues contain GAG chains,
usually of 5 to 15 kDa but occasionally much larger.
• Heparin is found in the granules of mast cells and also in liver, lung,
and skin.
• It is an important anticoagulant. It binds with factors IX and XI, but its
most important interaction is with plasma antithrombin.
• Can also bind specifically to lipoprotein lipase present in capillary
walls, causing a release of this enzyme into the circulation.
Heparan Sulfate
• Present on many cell surfaces as a proteoglycan and is extracellular.
• It contains Glucosamine with fewer N-sulfates than heparin, and,
unlike heparin, its predominant uronic acid is GlcUA.
• It is associated with the plasma membrane of cells, with their core
proteins actually spanning that membrane. In this, they may act as
receptors and participate in the mediation of the cell growth and cell -
cell communication.
• This proteoglycan is also found in the basement membrane of the
kidney along with type IV collagen and laminin, where it plays a major
role in determining the charge selectiveness of glomerular filtration.
 Dermatan Sulfate
• This substance is widely distributed in animal tissues. Its structure is
similar to that of chondroitin sulfate, except that in place of a GlcUA in
β-1,3 linkage to GalNAc it contains an IdUA in an α-1,3 linkage to
GalNAc.
• Formation of the IdUA occurs, as in heparin and heparan sulfate, by
5′-epimerization of GlcUA. Because this is regulated by the degree of
sulfation and because sulfation is incomplete, dermatan sulfate
contains both IdUA-GalNAc and GlcUA-GalNAc disaccharides.
• Dermatan sulfate has a widespread distribution in tissues, and is the
main GAG in skin. Evidence suggests it may play a part in blood
coagulation, wound repair and resistance to infection.
Functions of Proteoglycans
a. As a constituent of extracellular matrix or ground substance:
• Interacts with collagen and elastin
b. Acts as polyanions:
• Gylcosaminaglycans present in proteoglycans are polyanions and
 hence bind to polycations and cations such as Na and K. Thus attracts
water by osmotic pressure into extracellular matrix contributing to its
turgor.
c. Acts as a barrier in tissue:
• Hyaluronic acid in tissues acts as a cementing substance and
contributes to tissue barrier which permit metabolites to pass
through but resist penetration by bacteria and other infective
agents.
d. Acts as lubricant in joints:
• Hyaluronic acid in joints acts as a lubricant and shock absorbant.
Intraarticular injection of hyaluronic acid in knee joints is used to
alleviate pain in chronic osteoarthritis of knee joints.
e. Role in release of hormone:
• Proteoglycans like hyaluronic acid are present in storage or secretory
granules, where they play part in release of the contents of the
granules.
f. Role in cell migration in embryonic tissues:
• Hyaluronic acid is present in high concentration in embryonic tissues and
is considered to play an important role in cell migration during
morphogenesis and wound repair.
g. Role in glomerular filtration:
• Proteoglycans like hyaluronic acid is present in basement membrane (BM)
of glomerulus of kidney where it plays important role in charge-
selectiveness of glomerular filtration.
h. Role as anticoagulant in vitro and in vivo:
– In vitro.
• Heparin is used as an anticoagulant. 2 mg/10 ml of blood is used.
• Most satisfactory anticoagulant as it does not produce a change in red
cell volume or interfere with its subsequent determinations.
– In vivo.
• Heparin is an important anticoagulant.
• It binds with factor IX and XI, but its most important action is with
plasma antithrombin III. Binding of heparin to lysine residues in
antithrombin III produces conformational change which promotes the
binding of the latter to serine protease thrombin which is inhibited,
thus fibrinogen is not converted to fibrin.
• Four naturally occurring thrombin inhibitors in plasma are:
(i) Antithrombin III (75% of the activity)
(ii) α2-macroglobulin contributes remainder
(iii) Heparin cofactor II
(iv) α1-antitrypsin
• The last two shows minor activity.
i. Role as a coenzyme:
• Heparin acts in the body to increase the activity of the enzyme Lipoprotein
lipase. Heparin binds specifically to the enzyme present in capillary walls,
 causing a release of the enzyme into the circulation. Hence heparin is
called as Clearing factor.
j. As a receptor of cell:
• Proteoglycans like heparan sulphate are components of plasma
membrane of cells, where they may act as receptors and can participate
in cell adhesion and cell-cell interactions.
k. Role in compressibility of cartilages:
• Chondroitin sulphates and hyaluronic acid are present in high
concentration in cartilages and have a role in compressibility of cartilage
in weight bearing.
l. Role in sclera of eye:
• Dermatan sulphate is present in sclera of the eye where it has an
important function in maintaining overall shape of the eye.
m. Role in corneal transparency:
• Keratan sulphate I is present in cornea of the eye and lie between the
collagen fibrils. It plays an important role in maintaining corneal
transparency.