Biochemistry of Carbohydrates

Introduction
• Carbohydrates may be defined as polyhydroxy
aldehydes or ketones, or any substances that
yield one of these compounds on hydrolysis.
• Carbohydrates are the most abundant
biomolecules, distributed widely in plants and
animals where, they perform structural and
functional roles. They are hydrated carbon
molecules. Generally the hydrogen and oxygen
will be present in the proportion of 2:1.
• Many carbohydrates have the empirical formula
(CH2O)n, where n is 3 or larger.
• Some carbohydrates contain nitrogen,
phosphorous or sulphur also.
 BIOLOGICAL IMPORTANCE
• Carbohydrates provide the majority of energy in most organisms. They are
the most abundant dietary source of energy (4 Cal/g) for all organisms.
• Glucose is stored as glycogen in liver and muscle. Carbohydrates also
serve as the storage form of energy (glycogen) to meet the immediate
energy demands of the body.
• Carbohydrates (e.g. cellulose) give structure to cell walls (in plants) and
cell membranes.
• Carbohydrates serve as metabolic intermediates (e.g. glucose 6-
phosphate, fructose –1,6 bisphosphate). The metabolic intermediates
derived from glucose are used for the biosynthesis of amino acids, nucleic
acid and nucleotides.
• Carbohydrates (e.g. ribose, deoxyribose) comprise large portions of the
nucleotides that form DNA and RNA.
• Carbohydrates also play a role in lubrication, cellular intercommunication
and immunity.
• It plays an important role in the metabolism of proteins and fatty acids.
 Classification
• Based on the sugar units they contain, carbohydrates are classified
into four groups, as
– Monosaccharides
– Disaccharides,
– Oligosaccharides (a small polymer of sugar)
– Polysaccharides
• Monosaccharides are simple sugars, consisting of single
polyhydroxy aldehyde or ketone unit. They cannot be hydrolyzed to
yield simpler forms of sugar.
• They can be subdivided into trioses, tetroses, pentoses, hexoses,
heptoses and octoses, depending upon the number of carbon
atoms they possess.
• Carbohydrates with an aldehyde as their functional group are called
as Aldoses. Those with keto as functional group are called as
ketoses.
Structures of Aldoses & Ketoses
• Disaccharides yield two molecules of the same or different
monosaccharides when hydrolyzed. Examples are sucrose, lactose,
and maltose.
• Oligosaccharides Yield 3–6 monosaccharide units on hydrolysis. Eg.
maltotriose and raffinose
• Polysaccharides Yield more than 6 molecules of monosaccharide on
hydrolysis. Examples of polysaccharides are starch, cellulose,
glycogen and dextrins.
• The polysaccharides may be linear or branched eg. Cellulose is a
linear polysaccharide and starch is a branched polysaccharide.
• Polysaccharides are sometimes called as hexosans or pentosans,
depending upon the type of the monosaccharides they yield on
hydrolysis eg., glycogen -hexosan.
PHYSIOLOGICALLY IMPORTANT
MONOSACCHARIDES
 Derivatives of the Monosaccharides
• Many derivatives of the monosaccharides are found in the systems, which include sugar
phosphate, deoxy and amino sugars, sugar alcohol, and sugar phosphate, deoxy and amino
sugar, sugar alcohols, and sugar acids.
• Phosphate esters: When monosaccharides are used as fuel they are metabolized as
phosphate esters. For e.g., triose phosphate, ribose 5-phosphate and glucose 6-phosphate.
• Deoxy sugars: In these derivatives, a hydrogen atom replaces one of the hydroxyl groups
in the parent monosaccharide. An example is deoxyribose occuring in nucleic acid DNA, L-
Fucose (6-deoxy-L-galactose) is widely distributed in plants, animals and microorganisms. L-
rhamonose (6-deoxy-L-mannose) is also found as components of cell wall.
• Amino sugars: In a number of sugars, an amino group replaces one of the hydroxyl groups
in the parent monosaccharide. Some times the amino group is acetylated. Examples of
amino sugars are D-glucosamine, D-galactosamine and D-mannosamine. They commonly
occur in glycoconjugates. Several antibiotics contain amino sugars.
• Sugar alcohols: In sugar alchohols, the carbonyl oxygen of the parent monosaccharide has
been reduced producing a polyhydroxy alcohol. For example glycerol and myo-inositol are
the important components of lipids. Ribitol is a component of FMN and FAD.
• Sugar acids: Sugar acids are carboxylic acids derived from aldoses, either by the oxidation of
C-1 (the aldehyde carbon or by the oxidation of carbon bearing the primary alcohol). Sugar
acids are important components of many polysaccharides.
• Ascorbic acid or Vitamin C: It is derived from D-glucuronate . It is an essential cofactor for
many hydroxylation processes.
 REACTIONS OF MONOSACCHARIDES
Tautomerization or enolization:-The process of shifting a
hydrogen atom from one carbon atom to another to produce
enediols is known as tautomerization. Sugars possessing
anomeric carbon atom undergo tautomerization in alkaline
solutions.
When glucose is kept in alkaline solution for several hours, it
undergoes isomerization to form D-fructose and D-mannose.
This reaction known as the Lobry de Bruyn-von Ekenstein
transformation.
Reducing properties:- Benedict's test, Fehling's test,
Barfoed's test etc. The reduction is much more efficient in the
alkaline medium than in the acid medium.
The enediol forms or sugars reduce cupric ions (Cu2+) of copper
sulphate to cuprous ions (Cu+), which form a yellow precipitate
of cuprous hydroxide or a red precipitate of cuprous oxide.
0xidation:- Depending on the oxidizing agent used, the terminal
aldehyde (or keto) or the terminal alcohol or both the groups
may be oxidized.
1.Oxidation of aldehyde group (CHO ------>COOH) results in the
formation of gluconic acid.
2.Oxidation of terminal alcohol group (CH2OH ------>C OOH)
leads to the production of glucuronic acid.
Reduction:- When treated with reducing agents such as sodium
amalgam, the aldehyde or keto group of monosaccharide is reduced to
corresponding alcohol.

The important monosaccharides and their corresponding alcohols

are given below.
D-Glucose---------> D-Sorbitol
D-Galactose ------> D-Dulcitol
D-Mannose -----> D-Mannitol
D-Fructose ----> D-Mannitol + D-Sorbitol
D-Ribose -----> D-Ribitol
Sorbitol and dulcitol when accumulate in tissues in large amounts
cause strong osmotic effects feading to swelling of cells, and certain
pathological conditions. e.g. cataract, peripheral neuropathy,
nephropathy. Mannitol is useful to reduce intracranial tension by
forced diuresis.
 Dehydration:-

Osazone formation:-
Phenylhydrazine in acetic acid, when boiled with reducing sugars, forms osazones.
The first two carbons (C1and C2) are involved in osazone formation.
Glucose, fructose and mannose give the same type (needle-shaped) osazones.
Reducing disaccharides also give osazones maltose sunflower-shaped, and lactose
powderpuff shaped.
Formation of esters:- The alcoholic groups of
monosaccharides may be esterified by non-enzymatic or
enzymatic reactions and formation of esters.
GLYCOSIDES:- Glycosides are formed when the hemiacetal or
hemiketal hydroxyl group (of anomeric carbon) of a
carbohydrate reacts with a hydroxyl group of another
carbohydrate or a non carbohydrate (e.g. methyl alcohol,
phenol, glycerol). The bond so formed is known as
glycosidic bond and the non-carbohydrate moiety (when
present) is referred to as aglycone.
 Physiologically important glycosides
1.Glucovanillin (Vanillin-D-glucosides) is a
natural substance that imparts vanilla flavour.
2.Cardiac glycosides (steroidal glycosides):-
Digoxin and digitoxin contain the aglycone
steroid and they stimulate muscle contraction.
3.Streptomycin- An antibiotic used in the
treatment of tuberculosis is a glycoside.
4.Ouabain- Inhibits Na+ - K+ ATPase and blocks
the active transport of Na+.
 Structures
Racemic mixture : lf D- and L-isomers are
present in equal concentration, it is known as
racemic mixture or DL mixture. Racemic mixture
does not exhibit any optical activity, since the
dextro- and levorotatory activities cancel each
other.
D and L-isomers:-
The D and L isomers are mirror images of
each other. The spatial orientation of -H and
-OH groups on the carbon atom (C5 for
glucose) that is adjacent to the terminal
primary alcohol carbon determines whether
the sugar is D- or L-isomer.l f the -OH group is
on the right side, the sugar is of D-series, and if
on the left side, it belongs to L-series. The
structures of D- and L-glucose based on the
reference monosaccharide, D- and L-
glyceraldehyde (glycerose) are denoted in
Fig.2.1.
Epimers:- If two monosaccharides differ from each other in
their configuration around a single specific carbon (other than
anomeric) atom, they are referred to as epimers to each other.
Glucose and galactose are epimers with regard to carbon 4 (C4-
epimers). They differ in the arrangement of -OH group at C4.
Glucose and mannose are epimers with regard to carbon 2 (C2-
epimers).
Enantiomers:- Enantiomers are a special type of
stereoisomers that are mirror images of each other. The
two members are designated as D- and L-sugars.
Majority of the sugars in the higher animals (including
man) are of D-type.
 Linear & Cyclic form
A six-membered ring pyranose (based on pyran)
or a five-membered ring furanose (based on
furan). The cyclic forms of glucose are known as
α-D-glucopyranose and α-D-glucofuranos.
Anomers- The α and β cyclic forms of D-glucose are known as
anomers. They differ from each other in the configuration only
around C1 known as anomeric carbon. In case of α anomer, the -OH
group held by anomeric carbon is on the opposite side of the group -
CH2OH of sugar ring. The reverse is true for β –anomer. The
anomers differ in certain physical and chemical properties.
Mutarotation:- The α and β anomers of glucose have different
optical rotations. The specific optical rotation of a freshly prepared
glucose (α anomer) solution in water is +112.2° which gradually
changes and attains an equilibrium with a constant value of +52.7°.
In the presence of alkali, the decrease in optical rotation is rapid.
The optical rotation of β-glucose is +18.7°. Mutarotation is defined
as the change in the specific optical rotation representing the
interconversion of α and β forms of D-glucose to an equilibrium
mixture.
 Disaccharides
A disaccharide consists of two monosaccharide
units (similar or dissimilar) held together by a
glycosidic bond. They are crystalline, water-
soluble and sweet to taste.
The disaccharides 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.
 Maltose- is composed of two α-D-glucose units
held together by α (1-4) glycosidic bond. The free
aldehyde group present on C1 of second glucose
answers the reducing reactions besides the osazone
formations (sunflower-shaped).
Maltose can be hydrolysed by dilute acid or the
enzyme maltase to liberate two molecules of α-D-
glucose.
ln isomaltose, the glucose units are held together
by α (1-6) glycosidic linkage.
Cellobiose is another disaccharide, identical in
structure with maltose, except that the former has
β (1-4) glycosidic linkage. Cellobiose is formed
during the hydrolysis of cellulose.
Sucrose (cane sugar) is the sugar of commerce, mostly
produced by sugar cane and sugar beets. Sucrose is made
up of α-D-glucose and β-D-fructose. The two
monosaccharides are held together by a glycosidic bond
(α1- β2), between C1 of α glucose and C2 of β-fructose.
The reducing groups of glucose and fructose are involved
in glycosidic bond, hences ucrose is a non-reducing sugar,
and it cannot form osazones.
Sucrose is the major carbohydrate produced in
photosynthesis. It is transported into the storage organs
of plants (such as roots, tubers and seeds).
Sucrose is the most abundant among the naturally
occurring sugars. lt has distinct advantages over other
sugars as a storage and transport form.
This is due to the fact that in sucrose, both the functional
groups (aldehyde and keto) are held together and
protected from oxidative attacks.
 Sucrose is an important source of dietary carbohydrate.
lt is sweeter than most other common sugars (except
fructose) namely glucose, lactose and maltose. Sucrose
is employed as a sweetening agent in food industry. The
intestinal enzyme-sucrase-hydrolyses sucrose to glucose
and fructose which are absorbed.
Lactose is more commonly known as milk sugar since
it is the disaccharide found in milk. Lactose is
composed of β-D-galactose and β-D-glucose held
together by β (1-4) glycosidic bond. The anomeric
carbon of C1glucose is free, hence lactose exhibits
reducing properties and forms osazones (powder-puff or
hedgehog shape). Lactose of milk is the most important
carbohydrate in the nutrition of young mammals. It is
hydrolysed by the intestinal enzyme lactase to glucose
and galactose.
 Inversion of Sucrose:- Sucrose, as such is dextrorotatory
(+66.5°). But, when hydrolysed, sucrose becomes
levorotatory (-28.2°). The process of change in optical
rotation from dextrorotatory (+) to levorotatory (-) is
referred to as inversion. The hydrolysed mixture of
sucrose, containing glucose and fructose, is known as
invert sugar.
Hydrolysis of sucrose by the enzyme sucrase (invertasd or
dilute acid liberates one molecule each of glucose and
fructose. It is postulated that sucrose (dextro) is first split
into α–D-glucopyranose (+52.5°) and β-D-fructofuranose,
both being dextrorotatory. β-D-fructofuranose is less
stable and immediately gets converted to β-D-
fructopyranose which is strongly levorotatory (-92°). The
overall effect is that dextro sucrose (+66.5°) on inversion
is converted to levo form (-28.2°).
Structures
 Polysaccharides
Polysaccharide (simply glycans) consist of repeat units of monosaccharides or
their derivatives, held together by glycosidic bonds.
They are primarily concerned with two important functions-structural, and
storage of energy. Polysaccharides are linear as well as branched polymers. This
is in contrast to structure of proteins and nucleic acids which are only linear
polymers. The occurrence of branches in polysaccharidesis due to the fact that
glycosidic linkages can be formed at any one of the hydroxyl groups of a
monosaccharide.
Polysaccharides are of two types-
1.Homopolysaccharides: Which on hydrolysis yield only a single type of
monosaccharide. They are named based on the nature of the monosaccharide unit.
Thus, glucans are polymers of glucose whereas fructosans are polymers of
fructose.
2.Heteropolysaccharides: Which on hydrolysis yield a mixture of a few
monosaccharides or their derivatives.
Homopolysaccharides
Starch-
Starch is the carbohydrate reserve of plants which is the most
important dietary source for higher animals, including man. High
content of starch is found in cereals, roots, tubers, vegetables etc.
Starch is a homopolymer composed of D-glucose units held by α-
glycosidic bonds. lt is known as glucosan or glucan.
Starch consists of two polysaccharide components-water soluble
amylose (15-20 %) and a water insoluble amylopectin (80-85 %).
Chemically, amylose is a long unbranched chain with 200-1,000 D-
glucose units held by α (1--->4) glycosidic linkages.
Amylopectin is a branched chain with α (1---->6) glycosidic bonds at
the branching points and α (1--->4) linkages. Amylopectin molecule
containing a few thousand glucose units looks like a branched tree (20-
30) glucose units per branch).
Starches are hydrolysed by amylase (pancreatic or salivary) to liberate
dextrins, and finally maltose and glucose units. Amylase acts
specifically on α (1--->4) glycosidic bonds.
Structures
Dextrins-
Dextrins are the breakdown products of starch by the enzyme
amylase or dilute acids. Starch is sequentially hydrolysed through
different dextrins and, finally, to maltose and glucose.
The various intermediates (identified by iodine colouration) are
soluble starch (blue), amylodextrin (violet), erythrodextrin (red)
and achrodextrin (no colour).

Inulin-
Inulin is a polymer of fructose i.e., fructosan. It occurs in dahlia
bulbs, garlic, onion etc. lt is a low molecular weight (around
5,000) polysaccharide easily soluble in water. Inulin is not utilized
by the body. lt is used for assessing kidney function through
measurement of glomerular filtration rate (GFR).
Glycogen-
Glycogen is the carbohydrate reserve in animals, often
referred to as animal starch. It is present in high
concentration in liver, followed by muscle, brain etc.
Glycogen is also found in plants that do not possess
chlorophyll (e.g. yeast, fungi).
The structure of glycogen is similar to that of
amylopectin with more number of branches. Glucose is
the repeating unit in glycogen joined together by α (1-->4)
glycosidic bonds, and α (1-->6) glycosidic bonds at
branching points. The molecular weight (up to 108) and
the number of glucose units (up to 25,000) vary in
glycogen depending on the source from which glycogen is
obtained.
Structure of glycogen
Cellulose-

Cellulose occurs exclusively in plants and it is the most abundant

organic substance in plant kingdom. lt is a predominant constituent of
plant cell wall. Cellulose is totally absent in animal body.
Cellulose is composed of β-D-glucose units linked by β (1-->4)
glycosidic bonds.
Cellulose cannot be digested by mammals including man-due to lack of
the enzyme that cleaves β-glycosidic bonds (a amylase breaks bonds
only).
Ruminants and herbivorous animals contain microorganisms in the gut
which produce enzymes that can cleave β-glycosidic bonds. Hydrolysis
of cellulose yields a disaccharide cellobiose, followed by β-D-glucose.
Cellulose, though not digested, has great
importance in human nutrition. lt is a major
constituent of fiber, the non-digestable
carbohydrate. The functions of dietary fiber include
decreasing the absorption of glucose and
cholesterol from the intestine, besides increasing
the bulk of faeces.
Chitin:-
Chitin is composed of N-acetyl D glucosamine units
held together by β (1-->4) glycosidic bonds. lt is a
structural plolysaccharide found in the exoskeleton
of some invertebrates e.g. insects, crustaceans.
 Heteropolysaccharides
When the polysaccharides are composed of different types of
sugars or their derivatives, they are referred to as
heteropolysaccharides or heteroglycans.

MUCOPOLYSACCHARIDES-
Mucopolysaccharides are heteroglycans made up of repeating
units of sugar derivatives, namely amino sugars and uronic acids.
These are more commonly known as glycosaminoglycans (GAG).
Acetylated amino groups, besides sulfate and carboxyl groups
are generally present in GAG structure. The presence of sulfate
and carboxyl groups contributes to acidity of the molecules,
making them acid mucopolysaccharides.
Some of the mucopolysaccharides are found in combination with
proteins to form mucoproteins or mucoids or proteoglycans.
Mucoproteins may contain up to 95 % carbohydrate and 5 %
protein.
Mucopolysaccharides are essential components of tissue structure. The
extracellular spaces of tissue (particularly connective tissue-cartilage,
skin, blood vessels, tendons) consist of collagen and elastin fibers
embedded in a matrix or ground substance. The ground substance is
predominantly composed of GAG.
The important mucopolysaccharides include hyaluronic acid, chondroitin
4- sulfate, heparin, dermatan sulfate and keratan sulfate.
Hyaluronic acid:-
Hyaluronic acid is an important GAG found in the ground substance of
synovial fluid of joints and vitreous humor of eyes. it is also present as a
ground substance in connective tissues, and forms a gel around the
ovum. Hyaluronic acid serves as a lubricant and shock absorbant in
joints.
Hyaluronic acid is composed of alternate units of D-glucuronic acid and
N-acetyl D-glucosamine. These two molecules for disaccharides units
held together by β (1--->3) glycosidic bond. Hyaluronic acid contains
about 250-25,000 disaccharides unit (held by β 1--->4 bonds) with a
molecular weight up to 4 million.
Hyaluronidase:-
It is an enzyme that breaks (β 1--->4 linkages) hyaluronic acid and
other GAG. 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.

Chondroitin sulfates:-
Chondroitin 4-sulfate (Greek: Chondroscartilage) is a major constituent
of various mammalian tissues (bone, cartilage, tendons, heart, valves,
skin, cornea etc.).
Structurally, it is comparable with hyaluronic acid. Chondroitin 4-
sulfate consists of repeating disaccharide units composed of D-
glucuronic acid and N-acetyl D-galactosamine 4-sulfate.
Chondroitin 6-sulfate is also present in many tissues. As evident from
the name, the sulfate group is found on C6 instead of C4.
Structures of common glycosaminoglycans
Heparin:-
Heparin is an anticoagulant( prevents blood clotting) that
occurs in blood, lung, liver, kidney, spleen etc. Heparin
helps in the release of the enzyme lipoprotein lipase
which helps in clearing the turbidity of lipemic plasma.
Heparin is composed of alternating units of N-sulfo D-
glucosamine 6-sulfate and glucuronate 2-sulfate.
Dermatan sulfate:-
The name dermatan sulfate is derived from the fact that
this compound mostly occurs in the skin. lt is structurally
related to chondroitin 4-sulfate. The only difference is
that there is an inversion in the configuration around C5
of D-glucuronic acid to form L-iduronic acid.
Keratan sulfate:-
It is a heterogeneous GAG with a variable sulfate content, besides
small amounts of mannose, fructose, sialic acid etc. Keratan
sulfate essentially consists of alternating units of D-galactosamine
and N-acetylglucosamine 6-sulfate.
 BACTERIAL CELL WALL POLYSACCHARIDE
The cell walls of many bacteria are made of peptidoglycans, which are
heteroglycan chains linked to peptides(murein is another name for
peptidoglycan).
The heteroglycan component is composed of alternating residues of N-
acetylglucosamine and N-acetylmuramic acid joined by β(1→4)
linkage. N-acetylmuramic acid consists of N-acetylglucosamine in an
ether link with D-lactic acid.
Many such linear polymers lie side by side in the cell wall, cross-linked
by short peptides.
The antibacterial action of lysozyme (present in tears) is due to its
ability to catalyze the hydrolysis of polysaccharide chain of
peptidoglycan.
 Glycoproteins
Proteins are covalently bound to carbohydrates which are
referred to as glycoproteins.
The carbohydrate content of glycoprotein varies from 1 % to 90
% by weight.
The term mucoprotein is used for glycoprotein with
carbohydrate concentration more than 4 %. Glycoproteins are
very widely distributed in the cells and perform variety of
functions. These include their role as enzymes, hormones,
transport proteins, structural proteins and receptors.
The carbohydrates found in glycoproteins include mannose,
galactose, N-acetylglucosamine, N-acetylgalactosamine, xylose, L-
fucose and N-acetylneuraminic acid (NANA).
NANA is an important sialic acid.
 Functions of Glycoproteins
Antifreeze glycoproteins :-The Antarctic
fish live below -2 °C, a temperature at
which the blood would freeze. lt is now
known that these fish contain
antifreeze glycogtratein which lower
the freezing point of water and
interfere with the crystal formation of
ice. Antifreeze giycoproteins consist of
50 repeating units of the tripeptide,
alanine-alanine-threonin.
Blood group substances:- The blood
group antigens (of erythrocyte
membrane) contain carbohydrates as
glycoproteins or glycolipids. N-
Acetylgalactosamine, galactose, fucose,
sialic acid etc. are found in the blood
group substances. The carbohydrate
content also plays a determinant role in
blood grouping.
THANKS