Raffles Institution Carbohydrates 2020-2021

CORE IDEAS
(1) The Cell and Biomolecules of Life
Biological Molecules

CARBOHYDRATES
Content
The structure of carbohydrates and their roles in living organisms.

Learning Outcomes
(g) Describe the structure and properties of the following monomers :
(i) α-glucose and β-glucose (in carbohydrates)
(h) Describe the formation and breakage of the following bond:
(i) glycosidic bond (in carbohydrates)
(i) Describe the structures and properties of the following biomolecules and explain how these are
related to their roles in living organisms:
(i) starch (including amylose and amylopectin)
(ii) cellulose
(iii) glycogen

Other important learning outcomes not specifically mentioned in syllabus

Carry out tests for reducing and non-reducing sugars (including semi-quantitative use of the
Benedict’s test), and for starch (potassium iodide solution test)

References
Campbell, N.A. and Reece, J.B. (2011), Biology (9th edition), Pearson Benjamin-Cummings,
San Francisco
Taylor, D.J., Green, N.P.O., Stout, G.W. and Soper, R. (1997), Biological Science 1 (3rd
edition), Cambridge University Press, Cambridge
Brooker, R.J., Widmaier, E.P., Graham, L. and Stiling, P. (2008), Biology, McGraw-Hill, New
York

TABLE OF CONTENTS

(A) Introduction …………………..…………………………………………………………………….. 2

(B) Monosaccharides ………………………………………………………………………………….. 2
(C) Disaccharides ……………………………………………………………………………………… 5
(D) Polysaccharides……….…………………………………………………………………………… 9

* This handout is the effort of several Biology teachers at RI. It has and will continue to be
updated.
** Any information given in a double-lined box is for your information only.

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(A) INTRODUCTION

 Carbohydrates are organic compounds consisting of the elements carbon, hydrogen and
oxygen.
 Carbohydrates are carbon-containing compounds that are hydrated. Hence, their general
formula can be written as Cm(H2O)n, where m and n are variable whole numbers.
 They are the most abundant biomolecules on Earth. Most carbohydrates found in nature occur
as polysaccharides.

Carbohydrates

Simple Complex

Monosaccharides Disaccharides Oligosaccharides Polysaccharides

(1 monomer) (2 monomers) (3 -10 monomers) (>100 monomers)

(B) MONOSACCHARIDES
 Characteristics:
(a) Simplest and most basic form of carbohydrate Carbonyl group
= a carbohydrate which cannot be hydrolysed
to simpler carbohydrates
A hydroxyl group
(b) has a carbonyl group ( C = O)
(c) has multiple hydroxyl groups (-OH)
(d) general formula - (CH2O)n, where n = 3 to 7
(e) most names for monosaccharides end in –ose,
with pentoses (with 5-carbon atoms) and hexoses
(with 6-carbon atoms) being most common

 Classification:
3 different ways based on
(a) number of carbon atoms (size)
(range: 3 to 7 carbons long)
o triose (3-carbon)
o pentose (5-carbon)
o hexose (6-carbon)

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(b) location of carbonyl group

(i) aldose (aldehyde sugar) (ii) ketose (ketone sugar)
H R1

C O C O

R R2

Carbonyl group Carbonyl group

at the beginning of the is nested within the
carbon skeleton is carbon skeleton, and
called an aldo group. is called a keto
group.
This is an aldehyde
sugar or aldose. This is a ketone
sugar or ketose.

(c) spatial arrangement of atoms (or isomerism)

o linear and ring forms
- In an aqueous state, the linear forms of monosaccharides can bend to form a more
stable ring form. At any one time, most pentoses and hexoses exist in the ring form
(>99%); only a small portion of pentoses and hexoses exist in the linear form.
*Web link: See 3D model of glucose at http://www.biotopics.co.uk/JmolApplet/glucosejdisplay.html

Additional info: (doesn’t mean it won’t be tested as info here may help you answer questions)
Formation of the 6 carbon glucose ring

(a) Chemical equilibrium favours formation of rings. The open-chain form of glucose cyclizes when the O
of C5 attacks the C1 aldehyde group. Only the α-isomer is shown.
(b) Abbreviated ring structure. Carbons in the ring are omitted. Thicker edges indicate that you are
looking at the ring edge-on. Components attached to the ring lie above or below plane of the ring.

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o α- and β-forms
- When the linear carbon chain of glucose forms a ring, the hydroxyl group (-OH)
attached to carbon number 1 (C1) may be below or above the plane of the ring.
Hence the only difference between the α- and β-forms, is the position of hydroxyl
group at C1.

- In α - glucose, the (-OH) group attached to C1 is below the plane of the ring,
whereas in the β - glucose, the (-OH) group attached to C1 is above the plane of
the ring.
- The three forms of glucose (α-, β- isomers and linear forms) are interconvertible in
aqueous solution.

Additional info:
o isomerism - same molecular formula, different structure, different properties
- In this example, both have the same molecular formula (C6H12O6) but they are structurally
different. The slight difference in position of the hydroxyl group (-OH) at C4 is significant
enough to give the two sugars distinctive shapes and hence properties. Glucose is sweeter
than galactose. Glucose and galactose are isomers i.e. different compounds with the same
molecular formula.
1 1
* indicates the
2 2
carbon position
3 3 where there is a
* *
4 4 change in
5 5
orientation of the
hydroxyl group
6 6

 Significance of molecular structure of monosaccharides:

1 Small in size and has many hydroxyl groups that can form hydrogen bonds with water.
- readily soluble in water
- transported easily in water in animal and plant transport systems.

2 Linear form possesses a free carbonyl group (C=O)

- gives them reducing ability and hence are called reducing sugars.

3 Pentoses and hexoses can exist as rings.

- rings are stable building blocks for larger molecules (disaccharides and polysaccharides).

4 Ring structures exhibit α- and β-isomerism

- the same chemical formula gives rise to structurally different molecules.
- increases diversity of monosaccharides which can become building blocks for different
molecules.

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(C) DISACCHARIDES
 Characteristics:

(a) Disaccharides consist of two monosaccharides joined by a glycosidic bond or linkage.

A glycosidic bond is defined as a covalent bond formed between two monosaccharides by

condensation that involves the loss of one water molecule.

Conversely, the glycosidic bond can be broken by hydrolysis that involves the addition of
one molecule of water.

Removal of a water molecule Addition of a water

to synthesise a glyosidic bond molecule to break a bond
Condensation Hydrolysis

(b) have a general formula of Cn(H2O)n-1 (e.g. C12H22O11).

(c) each monosaccharide unit of a disaccharide is a monomer.

Disaccharide Monomers Remarks

maltose glucose + glucose sugar important in brewing beer
lactose glucose + galactose sugar present in milk
sucrose glucose + fructose sugar involved in transport in plants

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 Examples of disaccharide synthesis:

Maltose α-glucose + α-glucose A bond is formed between C1 of an α-glucose and C4
of the other α-glucose. The bond is thus called an
α(1-4) glycosidic bond.

4 1
+ H2O
3 2

α glucose α glucose maltose

Sucrose α-glucose + β-fructose A bond is formed between C1 of an α-glucose and C2

of the fructose. The bond is thus called an α(1-2)
glycosidic bond.

(For your info only: The β-position for fructose is defined as

the -OH being on the same side of the ring as the C6. The
α-position for fructose is defined as the -OH being on the
opposite side of the ring as the C6.)
6
1
5

4 1 2 5

+ H2O
3 2 3 4 6

α glucose

Exercise
1. Show how the hydrolysis of a glycosidic bond in a molecule of maltose which will produce 2
α-glucose molecules.

OH

Maltose

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Additional info:
 What is a reducing sugar?
o A reducing sugar “reduces” certain chemicals – the sugar acts as a ‘reducing agent’ by being
oxidised.
o In Benedict’s test, Cu2+ is reduced to Cu+ by a reducing sugar.
o Recall Benedict’s Test:
- tests for reducing sugar where the presence of a reducing sugar is indicated by a coloured
precipitate (green brick red with increasing concentration of reducing sugars in a solution).
o What makes a sugar reducing?
- reducing ability of aldose (only in the chain form) is due to the proton (-H) on carbonyl carbon
which can be oxidised to –OH. Thus, oxidation of aldose gives them the ability to reduce Cu(II)
to Cu(I).
- ketose cannot be oxidised directly (which means that they cannot act directly as a reducing
sugar). However, in the presence of a base (present in the Benedict’s reagent), ketose is
converted to aldose, which have reducing abilities.
E.g.: fructose is a ketose, and does not possess a proton at the carbonyl carbon. However in the
presence of the base in Benedict’s reagent, fructose isomerises into glucose or mannose (both
aldoses), which has reducing abilities.
- hence all monosaccharides will test positive in the Benedict’s Test.
H R1
.. base ..
aldose (aldehyde sugar) C O C O ketose (ketone sugar)
.. ..

R R2
Glucose Fructose Ring Open chain
CH2OH
OH
H
OH
O
OH

The residue at this end (label ‘ring’)

alternates between the ring form and open
1 chain form. When in open-chain form, the
2 carbonyl group is exposed for reducing
Both carbonyl
action. Likewise for lactose.
groups used in
formation of
glycosidic
bond

Lactose
NON-REDUCING REDUCING
(sucrose) (maltose, lactose)

 Examples of reducing and non-reducing sugars:

(a) Maltose is a reducing sugar because it has a free carbonyl group on one of the glucose
residues. Lactose is also a reducing sugar.
(b) Sucrose is a non-reducing sugar because it has no free carbonyl group. Both carbonyl
carbon groups of glucose and fructose are linked together during the formation of glycosidic
bond. Hence it loses its reducing function and is a non-reducing sugar.

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 Tests for reducing and non-reducing sugars:

Benedict's test: Test for reducing sugar (all monosaccharides and most disaccharides)
Principle Ability of reducing sugars to reduce Cu2+ to Cu+
 reducing ability of a reducing sugar is due to free carbonyl groups
 alkaline solution of copper (II) sulfate (CuSO4) [blue] is reduced to
insoluble copper (I) oxide (Cu2O) [red ppt]
heat

R C = O + 2CuSO4 + 4NaOH RCOOH + Cu2O + 2H2O + 2Na2SO4

H
Reducing sugar Benedict’s reagent Red ppt
(blue solution)

Cu2+ Cu+
Blue Red

Procedure (a) Place 2 cm3 of test solution in a test tube.

(b) Add equal volume of Benedict's reagent.
(c) Shake the mixture.
(d) Heat it by immersing the tube in boiling water bath for 3-4 minutes.

Observations and Presence of reducing sugar is indicated by a brick-red precipitate.

deductions Benedict’s test is partially quantitative.
Concentration of reducing sugar is determined by the amount of
precipitate formed which will determine the colour of suspension.
Overall colour suspension is determined by colour of ppt and the blue
colour of Benedict’s solution

[Reducing sugar] Increasing conc of reducing sugar   

Colour of Green Yellow Orange Brick red
suspension

Acid hydrolysis test, then Benedict’s test: Test for non-reducing sugar (eg: sucrose)
Principle Non-reducing sugar has to be hydrolysed into its reducing sugar
components by boiling with dilute hydrochloric acid first or by
digesting with hydrolytic enzymes.

Procedure If a negative result for Benedict’s test is obtained for the test solution,
then
(a) boil equal volume of a new sample of test solution with dilute
hydrochloric acid for about 1 minute to hydrolyse disaccharide to
monosaccharides,
(b) cool contents of tube,
(c) neutralise acidic content with sodium bicarbonate solution, and
(d) carry out Benedict's test for reducing sugar.
Observations and
Observation Deduction
deductions
A blue solution remains when Benedict’s test is first Non-
carried out. reducing
After acid hydrolysis, Benedict’s test is carried out sugars
again. present.
This time, a green/ yellow/ brown/ brick red ppt forms
in the blue solution

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(D) POLYSACCHARIDES
 Characteristics:
(a) Polysaccharides are formed by condensation of numerous monosaccharides.
Glycosidic bonds are formed between the monosaccharides, e.g. α(1-4), α(1-6), β(1-4)
etc.

The process of condensing many similar molecules to form a large molecule is called
polymerisation. Each monosaccharide component in the polymer is called a monomer.

(b) The general formula for polysaccharides is (C6H10O5)n where n ranges from 100s to 1000s.

 Classification:
Polysaccharides

Storage Structural

Starch Glycogen Cellulose

(α-glucose monomers) (α-glucose monomers) (β-glucose monomers)

Amylose Amylopectin

Polysaccharides can be classified according to their functions, either as storage (e.g. starch,
glycogen) or structural (e.g. cellulose) role.

(a) storage polysaccharides - starch

o starch is stored in plants as granules in chloroplasts and amyloplasts.
o enables plants to store surplus glucose which is a respiratory substrate oxidised
during cellular respiration to yield energy in the form of ATP (adenosine triphosphate).
Maltose is first released from this carbohydrate bank by hydrolysis of the glycosidic
bonds by enzyme amylase. Glucose is then released by hydrolysis of the dissacharide,
maltose, by enzyme maltase.
o starch from plants are made of 20% amylose and about 80% amylopectin.

Starch: the two forms of starch are amylose (unbranched) and amylopectin (branched). White ovals
in the micrograph are granules of starch within a chloroplast of a plant cell.

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Point of Amylose Amylopectin

Comparison
Monomer/ Made up of α-glucose monomers Made up of α-glucose monomers
branching forming unbranched polymers. forming branched polymers.

Bond(s) Glucose monomers are linked by Glucose monomers are linked by

α(1-4) glycosidic bonds. α(1-4) glycosidic bonds within a
branch and α(1-6) glycosidic bonds
at branch points.

No. of Each molecule consists typically of Each molecule contains up to 106

units/shape 200 to 20,000 glucose units. glucose monomers.

Each amylose molecule is coiled into Similar in structure to amylose

a helix as each residue is bent in (helix), except that there are helical
one direction with respect to the side chains attached at
previous residue. branchpoints (~every 20-30
glucose units) along each helix.

 test for starch:

Test for starch Iodine test (Iodine in potassium iodide test)
Principle  iodine (yellowish-orange) is not very soluble in water, therefore
iodine reagent is made by dissolving iodine in aqueous potassium
iodide. This makes a linear triiodide ion complex which is soluble.
 triiodide ion fits into centre of each turn of amylose helix, forming
a starch-iodine complex which gives a blue-black colouration.

Starch helix

Procedure Add a few drops of iodine solution to 1 cm3 of test solution (or a
piece of test specimen).

Observations Presence of starch is indicated by blue-black coloration.

and conclusion

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(b) storage polysaccharides – glycogen

o glycogen is an animal storage polysaccharide.
o structure very similar to that of amylopectin, except that it is even more extensively
branched.
o α-glucose monomers are linked by α (1-4) glycosidic bonds within a branch and α (1-6)
glycosidic bonds at the branch points. Branchpoints occur about every 12 glucose
monomers.
o each glycogen molecule is coiled into a helix as each residue is bent in one direction with
respect to the previous residue.
o humans and other vertebrates store glycogen mainly in liver and muscle cells. Hydrolysis of
glycogen in these cells gives glucose. Extensive branching allows for more ends for
enzymes to work on i.e. hydrolyse glycogen to form glucose.

(a) (b) (c)

Glycogen: (a) Extensive branching of glycogen. (b) Animal cells stockpile glycogen as granules
within liver and muscle cells. (The micrograph shows part of a liver cell.) (c) The extensively
branched structure of glycogen.

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 How do structures and properties of amylose, amylopectin and glycogen determine their
function as storage molecules?
1 All are made up of many glucose residues
 large energy store, can be hydrolysed ultimately to numerous monosaccharides
when required. Glucose is the major respiratory substrate to obtain ATP. Enzymes
(amylase) that hydrolyse these bonds are commonly available.
 large molecule that is insoluble in water, hence will not affect water potential (ψw)
of cells

2 All comprises of helices

 excellent for packing many subunits per unit volume,
i.e. compact molecule
 intramolecular hydrogen bonding and projection of
hydroxyl groups into the core of the helix formation.
 hence, relatively fewer hydroxyl groups are available for Intramolecular
hydrogen bonding with water making it insoluble in hydrogen
water. Will not affect ψw of cells. (note: starch and glycogen bonding
are hydrophilic and can still be hydrated but not dissolved).

3 Amylopectin and glycogen are branched

 multiple branch ends allow multiple hydrolytic enzymes (amylases) to work at the same
time i.e. hydrolyse amylopectin to two glucose units (maltose). Maltase further
hydrolyses maltose to glucose. Branching increases energy generation per unit time
(NOT per unit mass)

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(c) structural polysaccharides – cellulose

o cellulose is a plant structural polysaccharide.
o made up of β glucose monomers, linked via β(1-4) glycosidic bonds
o alternate β glucose monomers are rotated 180° (inverted) (NOT bond angle) with respect
to each other.
o Results in a linear, unbranched molecule with hydroxyl groups projecting out in both
directions.
o cellulose chains which are parallel to each other are held together by intermolecular
hydrogen bonds which form between these hydroxyl groups.
o many of these cross-linked linear cellulose molecules come together to form a microfibril. A
meshwork of such criss-crossing microfibrils form cellulose cell wall of plant cells.
o The cellulose cell wall
 has a porous structure due to gaps between microfibrils, making it freely permeable to
water and solutes. It allows free movement (DO NOT WRITE ‘allows regulation’) of
substances in and out of cells.
 is a strong and rigid structure as the meshwork distributes the stresses in all directions.
It serves to enclose plant cells and protect them from physical damage and bursting due to
osmotic stress.

β(1-4) glycosidic bonds between glucose monomers of a long, linear cellulose molecule.

o Structural organisation of cellulose cell wall

β-glucose cellulose microfibril

residue chain
(also called
molecule)
form β(14) -OH groups form
glycosidic bonds hydrogen bonds with parallel
with each other cellulose molecules

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o How do the structures in cellulose contribute to its properties and its function as a structural
molecule?

(a) As adjacent β glucose monomers are rotated 180o with respect to each other cellulose
molecules are linear.
The hydroxyl groups of each cellulose molecule forms hydrogen-bond with hydroxyl
groups of adjacent cellulose chains lying parallel to it forming cable-like units called
microfibrils.
 Hence microfibrils have high tensile strength.

(b) Microfibrils have relatively fewer hydroxyl groups available for hydrogen bonding with
water as only the surface of microfibril is exposed to water and many of OH groups are
already involved in forming hydrogen bonds with OH groups projecting from parallel
cellulose molecules. (note: cellulose is hydrophilic and can be hydrated but not dissolved)
 Hence microfibrils are insoluble in water.

Hence cellulose is a good structural polysaccharide as it can form microfibrils with high tensile
strength and that are insoluble in water.

Note: Cellulose is a food source for only certain bacteria, fungi, insects and mammals that have
genes that express the enzyme cellulase or live symbiotically with those that have. Cellulases
are not commonly available in organisms hence cellulose (compared to starch and glycogen) is
not readily hydrolysed as a respiratory substrate.

o Synthesis of cellulose molecules

Cellulose is synthesised by the enzyme, cellulose synthase. Cellulose synthase forms part
of a larger enzyme complex called the enzyme rosette that is embedded in the cell surface
membrane of plant cells. Bundles of cellulose microfibrils are laid down on the exterior of
the cell as the enzyme rosette moves along the membrane.

Q. Why is cellulose synthesised in the cell surface membrane and not inside the cell?

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Additional info:
More information about structural polysaccharides
(a) Chitin
: structural polysaccharide used by arthropods (insects, spiders, crustracean etc) to build their
exoskeletons
: fungi use chitin instead of cellulose as the building material for their cell walls
: similar to cellulose except that the glucose monomer of chitin has a nitrogen-containing appendage

Glucose monomer

Structure of chitin
monomer (N-acetyl-D- Glycosidic bonds between
glucosamine) chitin monomers
(b) Hemicellulose
: in contrast to cellulose that is crystalline, strong, and resistant to hydrolysis, hemicellulose has a
random, amorphous structure with little strength. It is easily hydrolysed by dilute acid or base, and
there are many naturally-occurring hemicellulases (enzymes) for its hydrolysis.
: residues of hemicellulose include many pentoses (e.g. arabinose and xylose), and hexoses (e.g.
mannose, galactose and glucose).

(c) Pectin
: acidic structural polysaccharides, found in fruit and vegetables and mainly prepared from 'waste'
citrus peel and apple pomace .(Pomace essentially consists of the pulp of the fruit after oil, water, or
other juice has been pressed out, the peel, seeds and stalks).

KEYWORDS
Some of the key words you should know include:

α, β glucose
Bonds: α(1-4) glycosidic bond, α(1-6) glycosidic bond, β(1-4) glycosidic bond
Branched chains
Cell wall
Compact
Condensation
Energy store/reserve
Enzymes
Helical/ helical coil
Hydroxyl groups
Hydrogen bond (with water)
Insoluble
Intermolecular/ interchain
Inverted / rotated 180o
Large molecules
Long straight/linear chains
Microfibrils
Parallel chains
Polysaccharides
High tensile strength
Water potential

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