Block 4

Unit 13 Amino Acids and Peptides
UNIT 13
Amino Acids and Peptides
Structure
13.1 Introduction 13.5 Structure of Peptides
Expected Learning Outcomes 13.6 Synthesis of Peptides
13.2 Structure and Physical Protection and Deprotection of
Properties of Amino Acids Amino Group
Structure of Amino Acids Protection and Deprotection of
Carboxy Group
Zwitterionic Nature
Peptide Bond formation using
Isoelectric Point and
Carboxy Activating Groups
Electrophoresis
13.7 Merrifield Solid-Phase
Stereochemistry of Amino Acids:
Synthesis
Optical Activity
13.8 Lab Detection of Amino Acids
13.3 Synthesis of 2-Amino Acids
13.9 Summary
Gabriel Phthalimide Synthesis
13.10 Terminal Questions
Strecker Synthesis
13.11 Answers
From 2-Halo Acids
13.4 Reactions of Amino Acids
13.1 INTRODUCTION
This is the first unit of Block 4 of this course. In this Unit, you will study about
the structure and chemistry of amino acids and peptides.
Amino acids are the compounds which contain both an amino group and a
carboxy group in their molecules. They constitute a particularly important class
of bifunctional compounds as they are the building blocks of proteins. Here,
you will first study about the amino acids and then the structure and synthesis
of peptides will be described.
We will begin this unit with a discussion on the structure and physical
properties of common amino acids. Then, we will explain the zwitterionic
nature of amino acids. This will be followed by description of the synthesis
of the amino acids.
Then, the structure of peptides will be illustrated. The synthesis of peptides

using N-protection, C-protection and C-activating groups will be discussed in
5
Block 4 Peptides, Proteins and Carbohydrates
detail. You will also study about Merrifield solid - phase synthesis which is a
versatile technique for the synthesis of peptides. Finally, the methods of
laboratory detection of amino acids will be explained.
Expected Learning Outcomes

After studying this unit and having the experiments performed, you should be
able to:
 list some amino acids and write their structures;
 give the common and IUPAC names of amino acids;
 discuss the zwitterionic nature of amino acids;
 explain the isoelectric point and importance of electrophoresis;
 give various methods of preparation of 2- amino acids;
 discuss the reactions of amino acids;
 briefly explain the structure of peptides;
 describe the methods of synthesis of peptides using N-protection,

C-protection and C-activating groups;
 discuss Merrifield solid-phase synthesis and highlight its importance;

and
 explain the laboratory detection of amino acids.
13.2 Structure and Physical Properties of Amino

Acids
Before studying the chemistry of amino acids, let us first understand their
structure.
13.2.1 Structure of Amino Acids

The general structure of a 2-amino acid can be represented as follows:
NH2
R CHCOOH
These are more often referred as α-amino acids.
While several hundred different amino acids are known to occur naturally, 20
of them deserve special mention as they are mainly present in proteins. These
amino acids are listed in Table 13.1. As given in the Table, for amino acids
trivial names are commonly used.
The convention to use a three letter code, as an abbreviation, for each amino
acid is also given in the table. These abbreviations are particularly useful in
designating the sequence of amino acids in peptides and proteins. The last
column of the table given below also shows a single letter code to denote the
amino acids. You will study more about peptides and proteins in Unit 14.
6
Table 13.1: Physical Properties of Amino Acids
Amino Acid, Name Abbreviation Abbreviation
NH2 Three letter One letter

Code Code
R CHCOOH
NH2
glycine Gly G
H CHCOOH
NH2
alanine Ala A
CH3 CHCOOH
CH3 NH2 valine Val V

CH3CH CHCOOH
CH3 NH2
leucine Leu L
CH3CHCH2 CHCOOH
CH3 NH2 Ile I

isoleucine
CH3CH2CH CHCOOH
NH2
methionine Met M
CH3SCH2CH2 CHCOOH
CH2
NH
CH2 proline Pro P
CHCOOH
CH2
NH2
CH2 CHCOOH phenylalanine Phe F
NH2
CH2 CHCOOH
tryptophan Trp W
N
H
NH2
serine Ser S
HOCH2 CHCOOH
OH NH2
threonine Thr T
CH3CH CHCOOH
NH2
cysteine Cys C
HSCH2 CHCOOH
7
NH2
HO CH2 CHCOOH tyrosine Tyr Y
O NH2
asparagine Asn N
H2NCCH2 CHCOOH
O NH2
glutamine Gln Q
H2NCCH2CH2 CHCOOH
O NH2
aspartic acid Asp D
HOCCH2 CHCOOH
O NH2
glutamic acid Glu E
HOCCH2CH2 CHCOOH
NH2
lysine Lys K
H2NCH2CH2CH2CH2 CHCOOH
NH2 NH2
arginine Arg R
H2NCNHCH2CH2CH2 CHCOOH
NH2
N
CH2 CHCOOH histidine His H
N
H
Amino acids can be classified as , , ,… etc., depending upon the location
of the amino group on the carbon chain containing the carboxy function. Some
examples are illustrated below:
2 1 3 2 1 4 3 2 1
H2 N CH2COOH H2N CH2CH2COOH H2N CH2CH2CH2COOH
2-aminoethanoic acid 3-aminopropanoic acid 4-aminobutanoic acid
(-amino acid) (-amino acid) (-amino acid)
Thus, the amino acids listed in Table 13.1 are -amino acids or 2-amino acids.
Now, answer the following SAQ.
SAQ 1
Give one example each for a 2-amino acid which contains the following in
the side chain:
i) Sulphur
ii) An aromatic ring
iii) A carboxyl group
8
13.2.2 Zwitterionic Nature

Because amino acids contain both carboxy and amino groups in their
molecules, they are amphoteric in nature, i.e. they behave both as acids and
bases. Amino acids actually exist as inner salts, called zwitterions. A
zwitterionic structure is possible for amino acids because the amino group is
basic in nature and can accept a proton from the acidic carboxy group. A
zwitterion can be represented as shown below.
COOH COO
H2 N H H3 N+ H
R R
a zwitterion
The highly polar nature of zwitterion allows the formation of strong crystal
lattices similar to the ionic compounds. Amino acids, therefore, resist
conversion from solid to liquid state and do not melt but decompose on
heating.
The zwitterionic nature is also reflected in their higher solubility in water and
low solubility in nonpolar solvents. In addition to the above observations, large
dipole moments also indicate the zwitterionic nature of amino acids.
13.2.3 Isoelectric Point and Electrophoresis

Let us now study the zwitterionic form of amino acids in more detail. You can
see in the zwitterion shown above that the amino group is protonated and the
carboxy group exists as the carboxylate anion. Thus, the acidic group is a
substituted ammonium ion and the basic group is the carboxylate anion. As a
result, in strongly acidic medium i.e., at low pH, the carboxylate group will be
protonated to yield the following species, I.
COO COOH
+
H
H3 N+ H H3 N+ H
+
H
R R
a zwitterion I
species present in
strongly acidic medium
Let us next consider the species present in strongly basic medium, i.e. at
higher pH of the solution. Under these conditions, the proton will be removed
from the +NH3 group to yield the following species.
COO COO
H+
H3 N+ H H2 N H
+ H+
R R
a zwitterion II
species present in
strongly basic conditions
9
Thus, we can write a combined equation for the acid-base behavior of the
acids as shown below.
COOH COO COO
H+ H+
H3 N +
H H3 N+ H H2 N H
+ H+ + H+
R R R
I a zwitterion II
low pH high pH
(acidic conditions) (basic conditions)
You can see that at low pH, species I has a net positive charge and has two
acidic sites (+NH3 and COOH). On the other hand, at high pH, species II has a
net negative charge and has two basic sites (NH2 and COO).
It is clear from the above equation that the amino group will be first protonated
and then the carboxylate anion. Also at some intermediate pH, the amino acid
exists as a zwitterion with no net charge. The pH at which this occurs is known
as isoelectric point, pHi of the amino acid. At this pH, the amino acid is
stationary in an electric field, i.e., it migrates neither to the negative pole nor to
the positive pole because the charges on it are balanced.
At low pH, there are two acidic sites in the amino acid I and therefore, it has
two pKa values. The pKa value corresponding to the more acidic site is referred
to as pKa1 and that corresponding to the less acidic site as pKa2. Thus, for the
simplest amino acid, glycine, we can write the two equilibria as follows:
+ pKa1 = 2.4 +
H3NCH2COOH + H2O H3NCH2COO + H3O+
+ pKa2 = 9.8
and H3NCH2COO + H2O H2NCH2COO + H3O+
At this stage you can compare the pKa1, with pKa of ethanoic acid which is
equal to 4.76. This leads to the conclusion that due to the electron withdrawing
nature of the protonated amino group, the acidity of amino acid is increased as
compared to ethanoic acid. Table 13.2 lists the pKa values and pHi of some
amino acids.
Table 13.2: pKa and pHi values of some amino acids

Amino acid pKa1 pKa2 pHi
Glycine 2.34 9.60 5.97
Alanine 2.34 9.69 6.00
Valine 2.32 9.62 5.96
Leucine 2.36 9.60 5.98
Isoleucine 2.36 9.60 6.02
Methionine 2.28 9.21 5.74
Proline 1.99 10.60 6.30
Phenylalanine 1.83 9.13 5.48
Tryptophan 2.83 9.39 5.89
Asparagine 2.02 8.80 5.41
Glutamine 2.17 9.13 5.65
Serine 2.21 9.15 5.68
Threonine 2.09 9.10 5.60
10
You can verify from Table 13.2 that

pKa1  pKa 2
pHi 
2
The amino acids having acidic and basic side chains are characterised by
three pKa values. The third pKa value, i.e., pKa3 reflects the nature of the
functional group present in the side chain.
Electrophoresis
It is a technique which involves the migration of charged particles using

an electric field. The charged particles have different mobilities
depending upon their shape, size and charge. Thus, different amino
acids and in turn, the peptides and proteins which are made up of these
amino acids, can be separated using electrophoresis. Since different
amino acids and peptides (or proteins) have different acidic and/or basic
groups, they have different isoelectric points. Thus, at a particular pH, in a
solution, they move towards the cathode or the anode at different rates.
In this way, electrophoresis can be used for the separation and analysis
of various amino acids, peptides or proteins when present in a mixture.
13.2.4. Stereochemistry of Amino Acids:

Optical Activity
With the exception of 2-aminoethanoic acid (glycine), the 2-amino acids have
at least one chiral centre.
According to the older D, L system of specifying the configuration (discussed

in Sec. 11.3, Unit 11, Block 3 of BCHCT-131 Course), the 2-amino acids
derived from animals or higher plants were found to have L configuration, i.e.,
they have the same relative configuration as L-glyceraldehyde. Thus, we can
write the following Fischer projection formula of an L amino acid.
COO Enzymes use up one

+ enantiomer
H 3N H preferentially.
R
an L amino acid
On the basis of your knowledge about assigning the absolute configuration,

can you attempt the following SAQ?
SAQ 2
What is the absolute configuration (R or S) of the following amino acids?
COO COO
+ +
a) H3 N H b) H3 N H
CH2OH CH2SH
L-serine L-cysteine
11
Having learnt about some general aspects of the structure of amino acids, let
us now focus our attention on the synthesis of 2-amino acids.
13.3 SYNTHESIS OF 2-AMINO ACIDS

2-Amino acids can be synthesised by using the methods given in Table 13.3.
Table 13.3: Methods of Synthesis of 2-Amino Acids
1. By N-alkylation of 1,2-benzenedicarboxylic imide (phthalimide) anion

O
COOCH2CH3 COOH
- +
NK +H C Br CH2 NH2
COOCH2CH3
O diethyl 2-bromopropanedioate
2-substituted amino acids can also be prepared.
2. From aldehydes : Strecker Synthesis
NH2
R CHO + NH3 + HCN R CH COOH
3. From 2-halo acids
X NH2
R CHCOOH + NH3 R CHCOOH
13.3.1 Gabriel Phthalimide Synthesis

This method is a modification of the Gabriel synthesis of amines which has
been discussed in Unit 11, Sec. 11.4.
It involves the N-alkylation of 1,2-benzenedicarboxylic imide (phthalimide)

anion with diethyl 2-bromopropanedioate as shown below:
O O
COOCH2CH3
COOCH2CH3
 +
HCBr + :N: K N CH
KBr
COOCH2CH3 COOCH2CH3
O O
diethyl 2-bromo- potassium 1,2-benzene- N-alkylated product
propanedioate dicarboxylic imide (85%)
(diethyl 2-bromo- (potassium phthalimide)
malonate)
The advantage of this method is that the alkylated product obtained in the
above reaction can be further alkylated to yield a variety of substituted amino
12 acids by the following sequence of reactions.
O O
COOCH2CH3  +
COOCH2CH3 Diethyl
1. CH3CH2O Na
N CH 2. RX N C R 2-bromopropanedioate
COOCH2CH3 COOCH2CH3 can be prepared by the
O O bromination of diethyl
propanedioate.
H+, H2O, D
2 CH3CH2OH
O O
H H COOH
+
H , H2O CO2
H 2N C R N C R N C R
COOH COOH
COOH
O O
2-amino acid
Thus, we can get a variety of amino acids depending upon the nature of R.
A variation of the above mentioned method utilises diethyl

2-(N-ethanoylamino)propanedioate instead of the imide derivative. The
sequence of reactions involved is shown below:
O COOCH2CH3 O COOCH2CH3
1. Na+ OC2H5
CH3CHNCH 2. RX CH3CHNCH C R
COOCH2CH3 COOCH2CH3
diethyl 2-(N-ethanoylamino)propanedioate
H+, H2O, D
O COOH
CH3COH + H2N C R
COOH
(not isolated)
CO2
H
H2 N C R
COOH
2-amino acid
13.3.2 Strecker Synthesis

It was pointed out in sub-Sec. 14.4.1, Unit 14, Block 3 of BCHCT-133 Course
that aldehydes on reaction with hydrogen cyanide yield a cyanohydrin. But,
when the same reaction is carried out in the presence of ammonia, the first
step is probably the initial formation of an imine from the reaction of the
aldehyde with ammonia. The addition of hydrogen cyanide to the imine
furnishes the corresponding 2-amino nitrile which on acidic or basic hydrolysis
yields the 2-amino acid. This is also known as Strecker synthesis. The
sequence of reactions involved in this synthesis is given below.
O NH NH2 NH2
NH3 HCN H+, H2O
CH3CH CH3CH CH3 C CN CH3CHCOOH
H2O
H
ethanal imine 2-aminopropane nitrile 2-aminopropanoic acid
(55%)
13
13.3.3 From 2-Halo Acids

In Unit 9, Sec. 9.5, you have studied that 2-halo acids can be obtained from
carboxylic acids using the Hell-Volhard-Zelinsky reaction. These 2-halo acids
on nucleophilic substitution by NH3 yield 2-amino acids as shown below:
H2O
CH3CHCOOH + 2 NH3 CH3CHCOOH + NH4Br
Br NH2
2-bromopropanoic acid 2-aminopropanoic acid

(65-70%)
Enzymes preferably It is also worthwhile to mention here that the amino acids obtained by
use one enantiomer synthesis using the methods discussed above are racemic mixtures.
Enantiomerically pure amino acids can be obtained by resolution of the
racemic mixtures or by chiral synthesis using enzymes.
SAQ 3
Which alkyl halide will you use for the synthesis of methionine in the Gabriel
phthalimide synthesis?
13.4 REACTIONS OF AMINO ACIDS

Amino acids undergo many of the reactions characteristic of the amino and
carboxy groups. For example, a typical reaction of carboxy group is
esterification and that of the amino functional group is alkanoylation. Let us
now study these reaction in detail.
1. Esterification
The carboxy group of an amino acid can be esterified in the normal way using
Methyl, ethyl and
benzyl esters are excess of an alcohol under acidic conditions.
used as intermediates
in the synthesis of + +
peptides. NH3 NH3
HCl
CH3CHCOO + CH3CH2OH CH3CHCOOCH2CH3Cl
(90-95%)
hydrochloride salt
of amino acid ester
Neutralisation of the hydrochloride with alkali yields the ester.
Esters of amino acids undergo intermolecular cyclisation to yield cyclic amides

shown below:
O H 2N O
C OC2H5 CH2 C NH
+ 2 C2H5OH CH2
CH2 C2 H5 O C CH2
O
NH C
NH2 O
2,5-diazacyclohexane-1,4-dione
(diketopiperazine)
14
2. Alkanoylation of Amino Acids
Alkanoylation of the amino group of an amino acid is carried out under basic
conditions so that the free amino form is present in substantial concentration.
Alkanoylation can be carried out by alkanoyl halides (acid chlorides) or
carboxylic acid anhydrides. The product is finally obtained by acidifying the
reaction mixture.
O
+
NH3 O NHCC6H5
 1. OH
(CH3)2CHCHCOO + C6H5CCl (CH3)2CHCHCOOH
H2O
valine benzenecar- 2 hr, 277 K N-benzenecarbonylvaline
bonyl 2. H+, H2O (N-benzoylvaline)
chloride (80%)
(benzoyl
chloride)
O O O
+
H3NCH2COO + CH3COCCH3 CH3CNHCH2COOH + CH3COOH
glycine ethanoic N-ethanoylglycine

anhydride (89-92%)
3. Reaction with Ninhydrin
When the aqueous solution of a 2-amino acid is treated with triketohydrindene

hydrate (ninhydrin), a blue-violet colour is obtained.
O
OH pH=9
2 + H3N+ CH CO2
OH
R
O
ninhydrin
O& O Ninhydrin test is given

by amino acids
N + R CH O + CO2 + H+ containing primary
amino group.
O O
Ruhemann's purple max = 570 nm
The blue-violet coloured compound formed is also known as Ruhemann’s

purple.
This is an important reaction used in the detection of small amounts of amino

acids.
4. Formation of Lactones
Some amino acids undergo cyclisation to yield cyclic amides, called lactams.
See Sec. 10.7, Unit 10 for nomenclature of lactams.
O
O O
+ D
H2NCH2CH2CH2CO H2NCH2CH2CH2COH :NH + HOH
H
(86%)
a lactam
15
5. Formation of Peptides
In addition to the above reactions, amino acids constitute the structural units of
peptides and proteins about which you will study in the forthcoming sections.
But before studying the formation of peptides, you must be curious to know
about the structure of peptides.
13.5 STRUCTURE OF PEPTIDES

Peptides are biologically important polymers in which 2-amino acids are joined
by the amide linkages, formed by the reaction of the carboxy group of one
amino acid with the amino group of another amino acid. These amide linkages
are also called peptide bonds. The general structure of a peptide is shown
The detailed structure
below:
of peptides will be 2
O R O
dealt in Unit 14. 12 o 153 pm
12
... NH C ...
1
C pm CH pm NH
5
o
114o 7 2
14 13
CH NH C CH
124 pm
123o 123o
1 3
R O R
peptide bonds
General Peptide Structure
Peptides can be classified as dipeptides, tripeptides and tetrapeptides,

depending on whether the number of amino acids is two, three or four,
respectively. Peptides containing up to 50 amino acids are called
polypeptides. Bradykinin is an important naturally occurring nonapeptide which
is present in blood plasma and is involved in the regulation of blood pressure.
Remember that a
three letter code to Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
represent the amino bradykinin
acids was given in
Table 13.1, Sec. 13.2
of this Unit. 13.6 SYNTHESIS OF PEPTIDES
The synthesis of peptides poses a challenge because even if you want to
synthesise a dipeptide from the starting amino acids A and B, we get a mixture
of following four dipeptides:
A-A, B-B, A-B and B-A
For example, the reaction of glycine and alanine will give the following four
The amino acids in a dipeptides:
peptide are also
known as amino acid Glycyl glycine , Alanyl alanine, Glycyl alanine and Alanyl glycine
residues.
Here, the convention is to write the N- terminal amino acid on the left and the
C- terminal amino acid on the right.
Therefore, for the selective synthesis of a particular peptide, we have to

16 protect
i) the N-terminal of one amino acid and
ii) the C-terminal of the other amino acid, and then
iii) combine or react the two protected amino acids to give the desired
dipeptide using the carboxy group activation .
activating group
protecting group
O O O O
X&NHCH &C& Z + H 2N&CHC &OY X &NHCH &C&NH &CH &C&OY+ HZ
R1 R2 R1 R2
protecting group amino acid 1 amino acid 2
peptide bond
dipeptide
The higher peptides can be synthesised by extending the chain of the

dipeptide either from the N-terminus side or the C-terminus side. For that we
have to deprotect and remove the protecting groups X or Y and repeat the
above sequence using the third amino acid.
Let us now understand the protection and deprotection of amino- and

carboxy- groups.
13.6.1 Protection and Deprotection of the Amino Group

The protection of amino group reduces its nucleophilicity. The following
reagents and groups are used for the protection of the amino group:
i) N-protection by Benzyloxycarbonyl and tert-Butoxycarbonyl

groups
Reagent Protecting group
O O
CH 2O&C&
CH 2OCCl
Phenylmethyl chloroformate Benzyloxycarbonyl

(Benzyloxycarbonyl chloride) (Cbz group or & Z group in older literature)
O O O
(CH3)3 COCOCC(CH 3)3 (CH3)3 COC &
tert-butoxycarbonyl
bis (1,1-dimethylethyl) dicarbonate
(Boc-group)
di-tert-butyl dicarbonate
The protection of the amino group of an amino acid by benzyloxycarbonyl

group can be represented as shown below:
O O
O
+ &
CH 2CCl + H 3NCHC &O 1. NaOH CH 2OC &NHCHCOOH
2. HCl, H2O R
R
III
(Z-protected amino acid)
(a carbamate) 17
Similarly, we can write for the protection by Boc group and get the
Boc- protected amino acid.
O
(CH 3)3COC &NHCHCOOH
R
(Boc-protected amino acid)
When required, after the formation of the required dipeptide, the deprotection
of the protected amino group can be carried out as follows:
1. By hydrogenolysis
The Boc benzyloxycarbonyl group can be removed from the Cbz- protected
amino acid by reaction with H2 in the presence of a transition metal catalyst.
The reaction gives toluene and the carbamic acid which on instantaneous
decarboxylation gives the unprotected amino group in the peptide.
CH 2H
O O
O O
H 2,Pd-C
CH 2OC &NH CHCONHCHCY
+ HOCNH & CHCONHCHCY + CO2
R1 R2
R1 R2
toluene
(Benzyloxycarbonyl protected peptide) O
carbamic
acid part H2N-CHCONHCHCY
R1 R2
2. By treatment with acid in mild conditions
The Boc carbamate group is stable in dilute base but it can be removed in mild
acidic conditions which do not affect the peptide bond. The deprotection of
Boc- protected amino group can be done by treatment with HBr in CH3COOH
or HCl.
Thus,
O
HBr +
(CH3)3COCNH - peptide CH2 = C(CH 3)2 + CO2 + H3N - peptide
CH3COOH
ii) N-protection by Phthaloyl (Phth) Group
The reaction of 1, 2-dicarboxylic acid anhydrides with amino acids yields

imides. Imides being stable to acids and hydrogenolysis, are good protecting
groups as they can be used in variety of synthetic conditions. Phthalic acid
derivatives can, thus, be used for N-protection.
Earlier, phthalic anhydride was used as phthaloylating agent which involved

fusion with the amino acid. These conditions were harsh and caused
racemisation. Even the use of solvents such as benzene, dioxane etc. could
not eliminate the problem of racemisation.
The protection of NH2 group can be done by the following phthaloylating

18 reagents:
O O
O
OH
N C OCH 2CH 3
OCH 2CH 3
O
O
monoethylphthalate
N-(ethoxycarbonyl) phthalimide
O
OCH 3
Cl P
OCH 3
O
3-chloro-3-(dimethoxyphosphoryl)isobenzofuran-1(3H)-one
The phthaloylation using the above reagents can be carried out under mild
conditions and the racemisation does not occur under these conditions. This is
shown below:
R R
O O
O
aq. Na 2CO 3
N C OCH 2CH 3 + o N COOH
H2N COOH 0 C,5 min or RT, 30 min
O O
Deprotection
The phthaloyl group can be removed by the following methods:
i) By hydrazinolysis
It involves treatment with hydrazine hydrate under reflux conditions

using methanol or ethanol as the solvent.
ii) Reductive opening of the ring followed by acid- catalysed lactonisation

of the formed hydroxyl compound.
R
R
O O
1. NaBH 4, isopropanol / H 2O, 24 hr NH CO

N
o
2. AcOH, 80 C, 2 hr
CH 2 OH
O
R
O
O + NH CO
2
CH 2
13.6.2 Protection and Deprotection of the Carboxyl

Group
The carboxyl group of the amino acids can be protected by converting it to an
ester. The methyl, ethyl or benzyl esters can be prepared. 19
Remember that methyl and ethyl esters can be conveniently prepared by

Fischer esterification.
O O
+
H ,D
H2N&CH &C&OH + R" OH H2N&CH &C&OR"
R' R'
R"= CH 3&, C2H5& ester
or CH 2 &
Deprotection: The ester group can be converted back to the carboxyl group
by the following methods at appropriate stage of the peptide synthesis.
i) Hydrolysis using aqueous base
O O
H2N&CH &C&OR" 1. aq. NaOH H2N&HC &C&OH + R"OH

+
2. H ,H2O R
R
But this method is not preferred because base can cause racemisation.
ii) The benzyl esters can be also deprotected by the following methods:
a) Hydrogenolysis with H2 using Pt or Pd as the catalyst, or
b) Treatment with HBr in acidic conditions in acetic acid.
13.6.3 Peptide Bond formation using Carboxy

Activating Groups
The most commonly used reagent for this purpose is dicyclohexylcarbodimide
(DCC).
DCC is the anhydride of N, N-dicyclohexylurea (DCU). DCC on reaction with

water gets converted to DCU
O
N C N + H2O NH C NH
1, 3-Dicyclohexylcarbodiimide N, N'-Dicyclohexylurea
DCC DCU
DCC activates the carbonyl group of the N-protected amino acid by taking
away a proton in the first step.
Step 1
O O
.. .. &
BocNH &CH &C&O &H + C6H11&N C N&C6H11 BocNH &CH &C&O
R R
+ ..
+ C6H11&N C N&C6H11
20 H
Step 2
In the next step, the carboxylate ion attacks the carbon of the C=N bond to
yield an O-acyl isourea.
C6H11
O O NH
& +
BocNH &CH &C&O + C6H11&N &
&C &
& N-C 6H11 BocNH &CH &C&O&C
H R
R N
C6H11
O-acyl isourea
Step 3
Nucleophilic addition of amino group of the carboxyl protected amino acid to

the carbonyl group of O-acyl isourea.
C6H11 C6H11
&
O NH O O NH
..
BocNH &CH &C&O&C + + H2N&CH &C&OR" BocNH &CH &C&O&C
+
R N R NH 2 N
R'
C6H11 CH-R' C6H11
O-acyl isourea
O C
OR"
tetrahedral intermediate
Step 4
Regeneration of carbonyl group and displacement of DCC as DCU.

C 6H 11
&
O NH O C 6H 11
BocNH &CH &C &O &C BocNH &CH &C NH
+
R HN H N R NH + O C
CH-R' C 6H 11 CH-R' HN
O C O C C 6H 11
OR" OR"
tetrahedral intermediate dipeptide DCU
The tetrahedral intermediate breaks down to give the dipeptide and

N, N’-dicyclohexylurea. Hence, we have synthesised a dipeptide which is
having both the amino end and the carboxy end protected with their respective
protective groups.
If we want of synthesise a tripeptide from this, we have to deprotect the amino

group and react it with another N-protected amino acid and repeat the rest of
the steps. Alternatively, to get a dipeptide from it, we can deprotect both the
protecting groups present at the amino end and the carboxy end. 21
Having understood the above sequence of steps to be followed for the

synthesis of a dipeptide, answer the following SAQ.
SAQ 4
Write the steps for the preparation of the peptide Gly-Ala.
SAQ 5
What could be the major problems associated with the above discussed
method of synthesis of peptides?
SAQ 6
Write all the possible structures of the dipeptides formed by the amino
acids glycine and valine.
13.7 MERRIFIELD SOLID PHASE SYNTHESIS

A major advancement in the syntheses of polypeptides was introduced by
R. Brace Merrifield in 1962. This technique is also known as polymer
supported synthesis and uses a polymer. Merrifield used a polystyrene in
which about 5% of the phenyl groups carried a chloromethyl (-CH2Cl) group in
their para positions. This functionalisation of the phenyl ring can be done by
Friedel -Crafts alkylation as given below:
Robert Bruce
Merrifield
(15th July, 1921- &CH &CH 2&CH &
14th May, 2006)
He was awarded ClCH 2OCH 2CH 3, SnCl 4

Nobel Prize in
Chemistry for the &CH 3CH 2OH
year 1984.
polystyrene
Polystyrene is a &CH &CH 2& CH &

polymer of styrene
which is
ethenylbenzene.
CH CH 2
CH 2Cl
functionalised polystyrene
The chloromethyl groups are reactive towards nucleophilic substitution

styrene reactions.
22 The solid phase synthesis of a dipeptide involves the following steps:

Step 1: The amino protected amino acid is attached to the polystyrene chain.
O R
&
(CH 3)3COC &NHCHCOO + Cl&CH 2 polystyrene chain
O R O
(CH 3)3COC &NHCHC &OCH 2 polystyrene chain
Step 2: The deprotection of the amino terminus is done by treatment with

trifluoroacetic acid.
O R O
(CH 3)3COC &NHCHC &OCH 2 polystyrene chain
CF 3COOH, CH 2Cl 2
R O
NH 2CHC &OCH 2 polystyrene chain
Step 3: Coupling with second N-protected amino acid is done using DCC.
The peptide (or amide) bond formation takes place in solution similar
to the previous method discussed in last sub-section. Here, the
difference is that the dipeptide formed is attached to the insoluble
resin.
R O O R'
NH 2CHC &OCH 2 polystyrene chain + (CH 3)3COC &NHCHCOOH
DCC
O R' O R O
(CH 3)3COCNHCHCNHCHCOCH 2 polystyrene chain + DCU
The DCU formed is removed by washing.
Step 4: Deprotection of the amino terminus of dipeptide
O R' O R O
(CH 3)3COCNHCHCNHCHCOCH polystyrene chain
2
CF 3COOH, CH 2Cl 2
R' O R O
NH 2CHCNHCHCOCH polystyrene chain
2
23
Step 5: If we want to stop at the dipeptide stage and do not wish to synthesise
higher peptides, the benzyl ester bond is broken using HF to release
the dipeptide from the polymer chain.
R' O R O
NH 2CHCNHCHCOCH polystyrene chain
2
HF
R' O R
+ &
H3NCHCNHCHCOO + FCH 2 polystyrene chain
dipeptide
Alternatively, if we wish to synthesise higher peptides, then coupling with third

N-protected amino acid is done using DCC .This is followed by deprotection of
the amino protected group, as explained above in Steps 3 and 4, respectively.
These steps, i.e. Steps 3 and 4, are repeated till we get the desired peptide;
and then finally, Step 5 is performed to get the product.
You have now studied two methods of synthesis of peptides. At this stage, you
may be curious to know which one is more advantageous. Obviously, the solid
phase synthesis has the following advantages over the previous method.
The polymer beads carrying the peptide chain are insoluble in the solvents
used in the synthesis. Hence, the product can be purified by simply filtering
and washing to remove the excess reagent and the byproducts.
In 1969, the enzyme ribonuclease was synthesised by Merrifield using solid

support. He designed a machine by which the steps required for the synthesis
could be performed automatically. This made the synthesis of peptides much
simpler and less time consuming.
The synthesis of ribonuclease required the addition of 124 N-protected

amino-acids and coupling reactions and 11,931 operations. The automated
machine made it possible in much shorter time and the intermediate stages of
isolation were also not required.
Similarly, synthesis of insulin involving 51 amino acids in two separate chains

comprising more than 5000 operations was achieved only in few days using
the automated procedure.
Automation in the protein synthesis has helped in the confirmation of the

structure of polypeptides obtained by degradation of chains. Also, new
proteins and polypeptides can be easily syntheised which are useful and have
better activity as compared to the ones available in nature. These new
synthetic options find applications in the field of medicine and biology.
13.8 LAB DETECTION OF AMINO ACIDS

The amino acids can be tested in the laboratory by the following methods.
1. Complexation with Cu2+ Ions
Aqueous solution of amino acids on addition of copper sulphate solution gives

24 a deep blue colour. The complex formed has the following structure:
H2N
O CO
RCH
Cu CHR
CO O
H2N
2. Ninhydrin Test
In Sec. 13.4 above, you have read about the reaction of amino acids with
ninhydrin.
Ninhydrin was accidentally discovered by Siegfried Ruhemann in 1910. He

was an German- English chemist .The reaction of amino acids and ninhydrin
was observed by him in the same year.
Ninhydrin is a white solid soluble in water, ethanol, and acetone etc. It reacts
with ammonia, primary and secondary amines, and peptides to form a blue-
purple coloured compound which is also known as Ruhemann's purple.
The mechanism of the formation of the blue colour is given below: Siegfried Ruhemann
th
4 January,1859–
O September, 1943)
O
C
C OH
&H2O N&CHCOOH
C
+ H2N&CHCOOH C This is a very
C R sensitive test and is
C OH also given by some
O
O β-amino acids
&CO 2
ninhydrin (3- amino acids) and
O some peptides as well
O
C as proteins, especially
H2O C on warming.
CH&NH 2 + CHO
CH &N CH
C
R C R
O
ninhydrin &H2O O Ninhydrin is also used
as a reagent for the
O O detection of latent
O& O fingerprints.
C C
+ C C
CH &N C &H
C &N C
C C
C C
O O
O O
blue-violet
13.9 SUMMARY
In this Unit, you have studied that
 20 amino acids occurring in proteins are L amino acids.
 The convention to use a three letter code, as an abbreviation, for each

amino acid. The single letter code are also used to denote the amino
acids.
 Amino acids exist as inner salts called zwitterions.
 At isoelectric point, pHi , the amino acid is stationary in an electric field,

i.e., it migrates neither to the negative pole nor to the positive pole
because the charges on it are balanced. 25
 2- Amino acids can be synthesised from potassium

1,2-benzenedicarboxylic imide, aldehydes (Strecker synthesis) and
2-halo carboxylic acids.
 The reactions of amino acids include the usual reactions of the carboxy
and the amino group. For example, they undergo esterification
(characteristic of the carboxy group) and alkanoylation (characteristic of
the amino group) reactions.
 Amino acids give a blue-violet color with ninhydrin.
 In peptides, 2-amino acids are joined by the amide linkages which are
formed by the reaction of the carboxy group of one amino acid with the
amino group of another amino acid. These amide bonds are called peptide
bonds.
 For the selective synthesis of a particular peptide, we have to protect i) the

N-terminal of one amino acid and ii) the C-terminal of the other amino acid,
and then iii) combine or react the two protected amino acids to give the
desired dipeptide using the carboxy group activation.
 Various protecting groups such as benzyloxycarbonyl, tert-butoxycarbonyl

and phthaloyl groups are used for protecting the amino group of the amino
acids.
 The polymer supported solid phase synthesis is advantageous because

the product can be purified by simply filtering and washing to remove the
excess reagent and the byproducts.
 Amino acids can be detected in the laboratory by complexation with

Cu2+ ions and ninhydrin test.
13.10 TERMINAL QUESTIONS

1. Which properties of amino acids indicate their zwitterionic nature?
2. Write the structures of leucine and isoleucine.
3. Name the amino acids which contain a basic group in the side chain.
4. Outline the Strecker synthesis of tyrosine.
5. Which protecting groups can be used for the protection of amino group of
the amino acid?
6. How can benzyloxycarbonyl group be removed from the N-

protected amino acid/peptide?
13.11 ANSWERS
Self-Assessment Questions
1. i) Methionine or cysteine
ii) Phenylalanine or Tyrosine or Tryptophan
26 iii) Glutamic acid or Aspartic acid

2. a) The order of priority of substituents is
+
H3N > COO > CH2OH > H
The given Fischer projection can be converted to another Fischer

projection given below by interchanging twice two substituents.
N + H3

OOC CH2OH
Overlooking H and moving from the substituents of highest priority

to lower priority, the direction is anticlockwise, so the configuration
is S.
b) Similarly, we can arrive at the configuration by following the steps

given in 2 a) above and get the configuration as R.
3. ClCH2CH2SCH3
O CH 3 O
4. (CH 3)3COCNHCH 2COOH + H2N CH &COCH 2C6H5 DCC
Boc &Gly Ala &OCH 2C6H5
O O CH 3 O
(CH 3)3COCNHCH 2CNHCH &COCH 2C6H5

Boc &Gly &Ala &OCH 2C6H5
+
1. H , H2O
2. H2, Pd&C
O CH 3
+ & + C6H5&CH 3 + CO2 + CH2 C(CH 3)2
H3NCH 2CNHCHCOO
Gly &Ala
5. It involves a large number of steps such as protection, activation,

coupling and deprotection. This is time consuming and involves
purification at each step. The overall yield obtained is also low.
6. i) C(CH 3)2 ii) H2N&CHCO&NH &CH2COOH

H2NCH 2CONH &CHCOOH C(CH3)2
iii) H2NCH2CONH&CH2COOH iv) H2N&CHCONH &CHCOOH

C(CH 3)2 C(CH 3)2
Terminal Questions
1. i) They form strong crystal lattices like ionic compounds.
ii) They decompose and not melt on heating. 27

iii) They are more soluble in water as compared to non-polar

solvents.
iv) They have large dipole moments.
2. Structures of leucine and isoleucine are given below.
CH3 NH2
CH3 NH2
CH3CH2CH CHCOOH
CH3CHCH2 CHCOOH
leucine isoleucine
3. Lysine, arginine and histidine
O NH2
NH3, HCN H+, H2O
4. HO CH 2CH HO CH 2CHCN
NH2
HO CH 2CHCOOH
tyrosine
5. Benzyloxycarbonyl, tert-butoxycarbonyl and phthaloyl groups are some

such protecting groups.
6. By hydrogenolysis i.e. reaction with H2 in the presence of a transition

metal catalyst.
28
Unit 14 Structure of Peptides and Proteins
UNIT 14
STRUCTURE OF
PEPETIDES
AND PROTEINS
Structure
14.1 Introduction Partial Hydrolysis
Expected Learning Outcomes End Group Analysis
14.2 Overview of Primary, 14.4 Summary
Secondary, Tertiary and 14.5 Terminal Questions
Quaternary Structures of
Proteins 14.6 Answers
14.3 Determination of Primary
Structure of Peptides and
Proteins
14.1 INTRODUCTION
In the previous unit, i.e. Unit 13, you studied about the chemistry of amino
acids as well as the structure and synthesis of peptides. We will further
elaborate on the structure of peptides and proteins in this unit.
Proteins are large polypeptides containing from about 50 to more than 8,000
amino acids per molecule. They have diverse biological functions. Proteins
catalyse and regulate various reactions occurring in our body as enzymes and
hormones. As skin and hair, they give outer covering to our body, and as
muscles they help in movement. In the form of antibodies, they protect us from
various diseases. The oxygen present in the air we breathe, is transported by
the protein hemoglobin. The nucleoproteins in the genes supply and transmit
the genetic information in cell division. In addition, proteins also provide
structural support in combination with other substances. After having an idea
about the importance of proteins, you must be curious to know about the
structure of peptides and proteins.
We will begin this unit with a discussion on the overview of structure of

peptides and proteins and explain different levels at which their structure is
understood. We will then elaborate on the details of the primary structure.
29
Here, we will keep our focus on the determination on the primary structure and
explain the methods used for this purpose.

able to:
 explain the primary, secondary, tertiary and quaternary structures of
peptides and proteins;
 explain the primary, secondary, tertiary and quaternary structures of
peptides and proteins;
 highlight the important features of α-helix and -pleated sheet
arrangements of amino acids in a polypeptide;
 describe the methods used for the determination of the primary structure
of peptides and proteins; and
 arrive at the primary structure of simple peptides from the data obtained
by their hydrolysis and end group analysis.
14.2 OVERVIEW OF PRIMARY, SECONDARY,

TERTIARY AND QUATERNARY
STRUCTURES OF PROTEINS
You are now aware about the importance of peptides and proteins. In this
section, you will study about what we broadly mean about their structure.
Since peptides and proteins contain a number of amino acids linked together, the
first step in the determination of their structure requires a knowledge about
which amino acids are present and how they are linked together as well as any
disulphide links present in them. The order in which the amino acids are joined is
The terms  and  called the primary structure.
refer to two
characteristic X-ray
Let us now study their secondary structure, i.e., the spatial arrangement of
diffraction patterns. the peptide chains.The description of the conformational relationship of the
The -type pattern nearest amino acids of a peptide is called its secondary structure. Two
was associated with conformational arrangements called α-helix and -pleated sheet are
right handed helix particularly stable. The -helix conformation is shown in Fig. 14.1.
and -type with the
pleated sheet.
In a right handed
helix, the chain turns
in clockwise direction.
30 Fig. 14.1: -helix.

In this the polypeptide chain forms a right handed coil having 3.6 amino acids
per turn. This allows hydrogen bonding between each carbonyl oxygen and an
amide hydrogen which stabilises the conformation. -Helix is important in
structural proteins such as -keratins which constitute proteins of skin, nails,
hair and feathers.
The -pleated sheet is shown in Fig.14.2 a). The adjacent chains in the
β-pleated structure may be aligned either parallel of anti-parallel to each other
as shown inFig.14.2 b).
a)
N-terminus N-terminus N-terminus C-terminus
NH 2 NH 2 NH 2 COOH
CH R CH R CH R R CH
C C O C N H
O O H
H N C
N N H O
R CH R CH R CH CH R
C C O C O H N
H O
H N
N H N C O
CH R CH R CH R R CH
C C O C N H
O O H
N H N C
N H O
R CH R CH CH R
R CH
C C O O H N
O C
H H
N C O
N H N
CH R CH R CH R R CH
C C O C N H
O O H
N H N C
N H O
C-terminus C-terminus C-terminus N-terminus
b)
Fig. 14.2: a) The -pleated sheet in a polypeptide and b) parallel and anti-
parallel arrangement of-pleated sheets
Note that the hydrogen bonds are formed between the carbonyl oxygen and
the amide hydrogen of two chains in -pleated sheet. You can see that the
side chains alternate above and below the planes of the pleated sheets.
In β-pleated sheet structure, there are two possibilities: parallel β-pleated

sheets or antiparallel β-pleated sheets. In parallel β pleated sheet
arrangement, polypeptide chains run in the same direction (i.e. from N- to
C-terminus) whereas in antiparallel β-pleated sheet, neighbouring chains 31
extend in opposite directions, see Fig.14.2 b). The hydrogen bonding present
in the two types of arrangements is different which makes antiparallel
arrangement more stable than the parallel one.
Tertiary structure refers to further folding of the peptide chain leading to its
three-dimensional shape. Folding affects the physical and biological
properties. Tertiary structure depends upon a variety of factors such as
hydrogen bonding, van der Waals forces and electrostatic forces. A protein
adopts the tertiary structure in such a way that favourable interactions are
maximised and the unfavourable ones are minimised.
The side chains of The tertiary structure of a protein is important in the sense that it defines an
some amino acids are active site wherein a substrate can bind (fit) as lock and key. This allows the
hydrophilic and in specificity of enzyme action. The tertiary structure of fibrous proteins show a
others, these are
super helix in which several -helices are coiled, see Fig. 14.3.
hydrophobic. In the
body, in an aqueous
solution, the
hydrophilic side
chains orient towards
the water while the
hydrophobic ones
turn away from the
water (towards inside
of the protein). These
interactions create
the looping and
folding of the protein.
Fig. 14.3: A super helix: helix within a helix.
Pronounced folding is observed in globular proteins in which the tertiary

structure allows them to be spherical. The globular proteins perform a crucial
Hemoglobin is
present in the red role in chemical transport.
blood cells and
The tertiary structure of a single polypeptide chain present in hemoglobin is
carries oxygen
throughout our body. illustrated below in Fig.14.4.
In sickle cell anemia,

a mutation results in
the change in one
amino acid in the
primary structure
which leads to
changes in the
secondary, tertiary Fig.14.4: Tertiary structure of a single polypeptide chain of hemoglobin.
and quaternary
structures of All proteins do not have a quaternary structure. Some proteins such as
hemoglobin. hemoglobin have a quaternary structure in which two or more peptide
chains combine to form an assembly. The manner in which these subunits are
organised is referred to as the quaternary structure of the protein.
Hemoglobin, contains four polypeptide chains where two chains are alpha and
two chains are beta chains. The quaternary structure of polypeptides present
32 in hemoglobin is shown below in Fig. 14.5.
Fig.14.5: Quaternary structure of polypeptides present in hemoglobin.
Photo Credits:
https://commons.wikimedia.org/wiki/File:1904_Hemoglobin.jpg
SAQ 1
The artificial sweetener, aspartame is as partylphenylalanine methyl ester.
Write its structure.
SAQ 2
What is meant by the primary structure of a peptide or a protein?
14.3 DETERMINATION OF PRIMARY

STRUCTURE OF PEPTIDES AND PROTEINS
Determination of the primary structure involves oxidation of the disulphide
bridges linking the chains in peptides or proteins to sulphonic acids using
peroxymethanoic acid, as shown below:
HCOOH
S S SO 3H HO 3S Disulphide links hold
peptide chains
together in a protein
molecule.
Chain A Chain B
Chain A Chain B
Proteins can be degraded into peptides by partial hydrolysis using either

dilute hydrochloric acid or enzymes. Peptides on complete hydrolysis by
heating with 6 N HCl for 24 hours yield a mixture of all the amino acids present in
it. This mixture is then separated, taking advantage of the acid-base properties
of amino acids, in an apparatus known as amino acid analyser. 33
The separation involves ion-exchange chromatography. This analysis, thus,

provides information about the amino acids present and their relative
amounts.
The sequence of amino acids present in a peptide chain can be determined either
by analysing the products of partial hydrolysis or by end group analysis. Let
us understand these methods with the help of examples.
14.3.1 Partial Hydrolysis
Frederick Sanger In a particular case, partial hydrolysis of a tetrapeptide containing Ala, Gly, Phe
(13th August 1918- and Val yielded a tripeptide Gly-Phe-Val and a dipeptide Ala-Gly.
19thNovember 2013)
hydrolysis
He was awarded two Tetrapeptide Gly Phe Val + Ala Gly
Nobel prizes in
Chemistry. One in the Since the dipeptide shows that Ala is linked to Gly, the amino acids in the
year 1958 and the tetrapeptide are linked in the following sequence:
other in 1980. He
shared the second Ala Gly Phe Val
Nobel prize
(a) (b)
with Walter Gilbert
(one-fourth to each),
for their contributions In the above structure, cleavage at (a) gives the tripeptide and that at (b)gives
related to the the dipeptide.
determination of base
sequences in nucleic 14.3.2 End Group Analysis
acids, and Paul
Berg (one-half) for his Let us first understand what is an end group? By convention, peptide
work on recombinant structures are written in such a way that the amino group is at the left and the
DNA. carboxy group is at the right. Thus, the amino end is called the N-terminus and
the carboxy end is called the C-terminus, the end groups being amino and
carboxy groups.
We will now discuss how to identify the N-terminus and C-terminus of a

peptide.
1. N-Terminal Identification by Degradation
The following two methods are used for this purpose:
i) Sanger Method
ii) Edman degradation
The amino groups of all the amino acids, except the N-terminal amino acid,
are involved in the amide bond formation. Therefore, the amino group of the N-
terminalamino acid is free and can react as a nucleophile.
The Sanger method of identifying N-terminal amino acid involves the reaction
of the free amino group in the peptide with l-fluoro-2,4-dinitrobenzene to yield
a peptide in which the N-terminal nitrogen is tagged with a 2,4-dinitrophenyl
group. After complete hydrolysis of the peptide, the tagged amino acid is
identified by chromatographic methods.
34 This procedure is illustrated below.

NO 2
O O O

O2 N F + H2NCHC NHCHC NHCH 2C NHCHCOO
CH CH 2C6H5 CH3
1-fluoro-2,4-dinitrobenzene
H3C CH3
ValPheGlyAla
Sanger utilised this

NO 2 method in the
O O O
determination of

O 2N NHCHC NHCHC NHCH 2C NHCHCOO sequence of amino
acids in insulin and
CH CH 2C6H5 CH3
was awarded Nobel
H3C CH3 Prize in 1958 for this
DNPValPheGlyAla
pioneering work.
Acid hydrolysis cleaves the amide bonds

of the 2,4-dinitrophenyl-labeled peptide, Insulin is a protein
giving the 2,4-dinitrophenyl-labeled H+, H2O which acts as a
N-terminal amino acid and a mixture
of unlabeled amino acids. hormone that
regulates sugar
NO 2 content in blood.
+ + +
NO 2 NHCHCOOH + H3NCHCOOH + H 3NCH 2COOH + H 3NCHCOOH
CH CH 2C6H5 CH3
H3C CH3
DNPVal Phe Gly Ala
The drawback of this method is that complete degradation is required after

the polypeptide is once tagged.
A more useful method is by the selective removal of the tagged terminal

amino acid and leaving the remaining chain intact so that it can be again
tagged with the reagent. One such method which enables identification of one
amino acid in the sequence at a time is the Edman degradation.
In Edman degradation, the terminal amino group adds to phenyl

isothiocyanate, C6H5N=C=S to yield a thiourea derivative. Treatment with mild
acid gives the tagged amino acid as a phenylthiohydantoin and the remainder
of the peptide chain. The tagged amino acid is identified and the remaining
peptide is again subjected to Edman degradation. This sequence is repeated
till all the amino acids in the peptide are identified. An example of Edman
degradation using the peptide Ala-Leu-Gly is shown below:
Edman degradation of a peptide

CH3 Pehr Victor Edman
th
(14 April 1916-
CH 2CHCH 3 H th
O H O 19 March 1977)
H2 N C C N C
C N C C OH
CH 3 H H O H H
Ala-Leu-Gly
peptide
35
+
B H
:
: :
O NH peptide
:S: C
: :
N C S
:O : C CHCH 3
+
:
N
:
N
:
H N CHCNH peptide
rest of peptide chain H H H
H CH3
B:
nucleophilic attack by amino group on isothiocyanate
with subsequent protonation of nitrogen deprotonation
+
H B
:O:
:
:
NH peptide
: :
:
O NH peptide
:S : C
:S : C +
CHCH 3 + H B
C
C CHCH 3
:
N N+
:
N N
H H
H H
cleavage of the peptide bond nucleophilic attack by sulphur
on carbonyl group
:O
: O O
:
:S  +
+ Cl H3NCHCNHCH 2COH + :B
+
CH3
:
N N CH2CHCH 3
H H CH3
LeuGly
unstable intermediate
D, rearrangement
S
N
N
O H
CH3
phenylthiohydantoin
derived from alanine
2. C-Terminal Identification with Carboxypeptidase Enzymes
The amino acid present at the C terminus is identified by enzymatic

hydrolysis. The enzymes which are used are called peptidases or
proteases. Thus, carboxypeptidases sequentially cleave the C-terminus
amino acids and the amino acids so cleaved are identified with time to know
the sequence.
Certain peptidases which allow controlled hydrolysis by cleaving certain

specific amide bonds, can also be used in sequence analysis. For example,
trypsin, present in intestine, catalyses the hydrolysis of the peptide bonds
involving carboxy groups of lysine or arginine. Similarly, chymotrypsin allows
cleavage of the amide bonds of amino acids containing aromatic groups in
their side chains, namely phenylalanine, tyrosine and tryptophan.
36
SAQ 3
Write the structure of the phenylhydantoin formed by valine when present in
the dipeptide Val-Gly.
SAQ 4
A tripeptide contains the following sequence of amino acids:
Met-Leu-Phe.
Which reagent will you use for identification of amino acid present at its
C-terminus?
14.4 SUMMARY
In this Unit, you have learnt that
 The order in which the amino acids are joined is called the primary structure.
 The description of the conformational relationship of the nearest amino

acids of a peptide is called its secondary structure.
 Two conformational arrangements called α-helix and -pleated sheet are

particularly stable.
 Tertiary structure refers to further folding of the peptide chain leading to its
three-dimensional shape. Folding affects the physical and biological
properties.
 The manner in which two or more peptide chains combine to form an

assembly is called the quaternary structure of the protein.
 Proteins can be degraded into peptides by partial hydrolysis using either

dilute hydrochloric acid or enzymes.
 Peptides on complete hydrolysis by heating with 6 N HCl for 24 hours yield a

mixture of all amino acids present.
 The sequence of amino acids present in a peptide chain can be determined

either by analysing the products of partial hydrolysisor by end group
analysis.
 The following methods are used for N-Terminal identification by

degradation:
 Sanger Method
 Edman degradation
 The amino acid present at the C terminus is identified by enzymatic

hydrolysis. The enzymes which are used are called peptidasesor
proteases. 37

1. A heptapeptide contained three units of alanine, two units of glycine and
one unit each of serine and valine. It yielded the following mixture of
tripeptides and dipeptides on partial hydrolysis:
Ala-Gly-Ala, Ala-Val-Ser, Ser-Ala and Ala-Gly
What is the primary structure of the heptapeptide?
2. Which type of structure governs the overall the three-dimensional folding of

a polypeptide?
3. Why are globular proteins spherical in shape?
4. Why do some proteins have quaternary structure while the other do not
have?
5. How is the degradation of a polypeptide by Sanger method different from

the Edman degradation?
6. Illustrate different levels of structure of polypeptides and proteins using a

diagram.
14.6 ANSWERS
Self Assessment Questions
1. It is a dipeptide with the methyl ester. So the structure is
O NH2 O O
HOCCH2 CH C NH CH C O CH3
aspartic acid part CH2
methyl ester
phenylalanine part
2. The sequence of amino acids present in a peptide or protein indicates its

primary structure.
3. Phenylhydantoin formed by valine has the following structure:
S
N
N
O H
CH
CH 3 CH 3
38 4. Chymotrypsin
Terminal Questions
1. Ala-Gly-Ala-Val-Ser-Ala-Gly or Ala-Val-Ser-Ala-Gly-Ala-gly
2. Tertiary structure
3. Pronounced folding in globular proteins due to their tertiary structure make

them spherical.
4. Because quaternary structure of a protein results only when two or more

peptide chains are present in it.
5. In Sanger method, we can identify the amino acid present at N-terminal of

the polypeptide whereas in Edman degradation, the sequence in which the
amino acids are present can be identified.
6.
Photo Credit: https://commons.wikimedia.org/wiki/File:225_Peptide_Bond-01.jpg
39
UNIT 15
CARBOHYDRATES-I:
MONOSACCHARIDES
Structure
15.1 Introduction Absolute Configuration of
Glucose and Fructose
Mutarotation
15.2 Classification of
Carbohydrates 15.6 Ascending and Descending
of Chains in
15.3 General Properties
Monosaccharides
15.4 Structure of Glucose and
15.7 Summary
Fructose
15.8 Terminal Questions
15.5 Configuration of
Monosaccharides 15.9 Answers
15.1 INTRODUCTION
In the last two units of this Block, i.e. Units 13 and 14, you have studied about
the chemistry of amino acids and structure of peptides and proteins.
In this unit and in the next unit, you will study about another such category of
compounds viz. carbohydrates. Carbohydrates, proteins, nucleic acids and
fats occur in almost all organisms and play an important and primary role in
metabolic processes. These natural products are called primary metabolites.
We will begin this unit with a discussion on the importance and classification of
carbohydrates which will give you an idea about their vast variety and diverse
roles. Here, in this unit, we will keep our focus on monosaccharides only and
discuss about disaccharides and polysaccharides in the next unit. The general
properties including the reactions exhibited by monosaccharides will be
explained. Then, we will describe how the structures of monosaccharides are
represented. We will specifically elaborate on the structures and absolute
configuration of the familiar monosaccharides-glucose and fructose.
Finally, the methods of ascending and descending of carbon chains in

40 monosaccharides will be discussed.
Unit 15 Carbohydrates - I: Monosaccharides

able to:
 classify a carbohydrate as a monosaccharide, oligosaccharide

(disaccharide, trisaccharide and so on) or a polysaccharide;
 give some examples of monosaccharides and classify them as an

aldose or a ketose;
 explain some general reactions exhibited by monosaccharides;
 write the structures of various monosaccharides in different

representations such as Fischer projections and Haworth projections;
 explain mutarotation; and
 discuss the methods of ascending and descending of carbon chains in

monosaccharides.
15.2 CLASSIFICATION OF CARBOHYDRATES

Carbohydrates received their name because of their general formula
Cx(H2O)y, according to which they appear to be hydrates of carbon.
Carbohydrates are wide spread in nature. In plants, they constitute up to80%

of the dry weight. Carbohydrates occurring in plants include cellulose (which
gives structural support to plants), starch (which serves as the reserved
energy source) and sugars (like sucrose and glucose). Sugars are
crystalline substances
Glucose is an essential constituent of blood in higher animals and occurs in having sweet taste
polymeric form as glycogen, in liver and in muscles. Carbohydrates also occur and are soluble in
water.
in adenosine triphosphate which is involved in biological energy storage and
transport systems. They are also present in the nucleic acids which control the
production of enzymes and the transfer of genetic information.
In nature, carbohydrates are synthesised by a process called photosynthesis.

In this process, sunlight impinging on chlorophyll present in the green plants is
absorbed and the photochemical energy, thus obtained is used to convert carbon
dioxide and water into carbohydrates and oxygen. The overall process can be
represented as follows:
sunlight, chlorophyll
x CO 2 + y H2O Cx(H2O)y + x O 2
Carbohydrates are polyhydroxy aldehydes and ketones and substances which

hydrolyse to polyhydroxy aldehydes and ketones.
The simplest carbohydrates are called sugars or saccharides, (Latin:

Saccharum, sugar). Carbohydrates can be classified as monosaccharides,
oligosaccharides and polysaccharides.
Let us now study about the monosaccharides in detail.

41
Monosaccharides
Monosaccharides are the simplest carbohydrates which cannot be hydrolysed

into smaller and simpler carbohydrates. A monosaccharide may be further
classified as an aldose or a ketoseif it contains an aldehydeor a ketogroup,
respectively. The simplest monosaccharides are 2,3-dihydroxypropanal, an
aldose(glyceraldehyde)and1,3-dihydroxypropanone, a ketose. Their structures
are as shown below:
CHO CH 2OH
H C OH C O
CH 2OH CH 2OH
2,3-dihydroxypropanal 1,3-dihydroxypropanone
(an aldose) (a ketose)
Depending upon the number of carbon atoms in the chain, sugars are called
trioses (3 carbons), tetroses(4 carbons), pentoses(5 carbons), hexoses(6
carbons) and so on. Therefore, 2, 3-dihydroxypropanal is an aldotrioseand 1, 3-
dihydroxypropanone is a ketotriose.
It was pointed out in Sec.11.3, Unit11 of Block 3 of BCHCT-131 Course that

monosaccharides can be classified as D or L depending upon whether the
position of the hydroxyl group on carbon next to primary alcoholic group is right
or left in the Fischer projection of the molecule projected vertically in such a
way that the aldehyde function is on the top. The structures of aldoses
belonging to D-family are given below in Fig.15.1.
CHO
Most of the naturally
occurring sugars H C OH
belong to D family. CH 2OH

D-(+)-glyceraldehyde
Remember that (+)
and () signs indicate CHO CHO
the optical activity. H C OH HO C H
H C OH H C OH
CH 2OH CH 2OH
D-(-)-erythrose D-(-)-throese
CHO CHO CHO

CHO
OH HO C H H C OH HO C H
H C
H C OH H C OH HO C H HO C H
H C OH H C OH H C OH H C OH
CH 2OH CH 2OH CH 2OH CH 2OH

D-(-)-ribose D-(-)-arabinose D-(+)-xylose D-(-)-lyxose
CHO CHO CHO CHO CHO CHO CHO CHO
H C OH HO C H H C OH HO C H H C OH HO C H H C OH HO C H
H C OH H C OH HO C H HO C H H C OH H C OH HO C H HO C H
H C OH H C OH H C OH H C OH HO C H HO C OH HO C H HO C H
H C OH H C OH H C OH H C OH H C OH H C OH H C OH H C OH
CH 2OH CH 2OH CH 2OH CH 2OH CH 2OH CH 2OH CH 2OH CH 2OH

D-(+)-allose D-(+)-altrose D-(+)-glucose D-(+)-mannose D-(-)-gulose D-(-)-idose D-(+)-galactose D-(+)-talose
42 Fig.15.1: The D-family Aldoses

Each of the D sugars shown above has an enantiomeric L-counterpart.
As in the case of D-aldoses shown in Fig.15.1, the Fischer projections

of monosaccharides belonging to D-ketose series are shown in
Fig.15.2.
CH 2OH
O
CH 2OH
1,3-dihydroxypropanone
CH 2OH
O
H OH
CH 2OH
D-(-)-erythralose
CH 2OH CH 2OH
O O
H OH HO H
H OH H OH
CH 2OH CH 2OH
D-(+)-ribulose D-(+)-xylulose
O O O O
H OH HO H H OH HO H
H OH H OH HO H HO H
H OH H OH H OH H OH

D-(+)-psicose D-(-)-fructose D-(+)-sorbose D-(-)-tagatose
Fig.15.2: The D-ketoses
Having seen the large variety in the structures of monosaccharides, let us

understand their general properties.
But before that answer the following SAQ to check your understanding.
SAQ 1
Fill in the blanks in the following with the words given below:
five, pentose, hexose, four, ketose, aldose
i) Ribose is a…………..while fructose is a …………….
ii) Xylulose is a …………having a ……….carbon chain.
iii) D (-)-erythrose is a………having…….. carbon atoms in the chain.
43
15.3 GENERAL PROPERTIES

Tollens reagent is
freshly prepared by first Monosaccharides are colourless crystalline solids which are soluble in water.
adding a few drops of The exhibit the following reactions:
dilute NaOH and then
aqueous ammonia to Reactions of Monosaccharides
aqueous silver nitrate
solution. Initially, the
1. With Tollens reagent- containing [Ag+(NH3)2–OH]. Tollens reagent is
brown solid silver oxide reduced by aldoses and α-hydroxyketones. The corresponding
forms which dissolves on monosaccharide gets oxidised to the aldonic acid. The silver formed is
addition of aqueous this reaction gets deposited on the walls of the test tube forming a silver
ammonia to yield
mirror.
diammine silver (I)
+
complex, [Ag (NH3)2] in
2. With Fehling’s Solution (which contains a cupric tartrate complex in
the clear solution.
alkaline solution) and Benedict’s reagent (which contains a cupric ion
complexed with citrate ion).
Fehling’s solution A is
made by dissolving Aldoses and ketoses (α-hydroxyketones) reduce both Fehling’s solution
69.28 g CuSO4. 5H2O in
water and making
and Benedict’s solution. Hence, the inital blue colour of the complex
solution to 1 litre volume. present in these solutions changes to brick red due to the formation of
Cu2O. In alkaline medium, aldoses undergo isomerisation reactions and
Fehling’s solution B is
prepared by dissolving equilibrium exists between an aldose, enediol form, ketose and isomeric
sodium potassium (epimeric) aldose.
tartrate,
H O H OH OH
C4H4O6NaK.4H2O,
Rochelle salt (346g) and C C H2C
sodium hydroxide (120

g) in water and making H C OH C OH C O
the solution up to 1 litre
volume. H (OH)n (H OH ) n (H OH)n
Both Fehling’s solution A CH3 CH2OH

CH2OH
and B are mixed in equal
ketose
volume and used as aldose enediol
intermediate
Fehling’s solution.
Benedict’s reagent is a H O
solution of copper C
sulphate pentahydrate,
sodium hydroxide and HO C H
tartaric acid.
(H OH)n
Benedict’s reagent
contains copper sulphate CH2OH
(173 g), sodium epimeric aldose
carbonate (100 g) and
sodium citrate (173 g) in Carbohydrates which contain the hemiacetal group give positive Tollens
1000 ml of solution in and Benedict’s tests and hence, these are called reducing sugars.
water. Sodium
carbonate keeps the This is so because in the aqueous solution, there is an equilibrium
solution alkaline while
between the hemiacetal and the open chain structure of the sugar. It is
sodium citrate i.e. citrate
ions, complex with Cu the open chain form which gets oxidised and the equilibrium keeps on
(II) ions and stop their shifting to give more of the open chain form.
conversion to Cu (I) ions
on storage. However, formation of an acetal linkage does not allow such equilibrium
and those sugar glycosides which contain acetal linkages instead of
hemiacetals do not give positive tests with Tollens’ reagent and Benedict’s
44 reagent are called non-reducing sugars.
You will study more about reducing and non-reducing nature of sugars
when you study the structures of disaccharides in Unit 16.
3. Reaction with Bromine Water
Since bromine water is mildly acidic with pH 6.0, no isomerisation takes

place when a monosaccharide is oxidised with it to yield an aldonic acid in
which only the -CHO group gets oxidised to -COOH group.
CHO COOH
Br2
(CHOH)n (CHOH)n
H2O
CH2OH CH2OH
an aldose an aldonic acid
(glyconic acid)
4. Reaction with Nitric Acid
The reaction of an aldose with dil. nitric acid, which is a stronger oxidising
agent than bromine water, oxidises both -CHO and -CH2OH groups and
yields an aldaric acid as the product.
CHO COOH
HNO3
(CHOH)n (CHOH)n
CH2OH COOH
an aldaric acid a dicarboxylic acid
5. Reduction of Monosaccharides
Both aldoses and ketoses can be reduced using sodium borohydride or H2

and Pt catalytic reduction to the corresponding alcohols.
CHO CH2OH
NaBH4
(CHOH)n (CHOH)n
CH2OH CH2OH
an aldose an alditol
The alditol formed by D-glucose is called D-glucitol, commonly also known

as D-sorbitol. Can you write its structure?
Yes, it is like this.

CH2OH
H OH
HO H
H OH
H OH
CH2OH
D-glucitol
(D-sorbitol) 45
6. Reaction with Phenylhydrazine
You have already studied in your second semester course, BCHCT-133,

that aldehydes react with phenylhydrazine to give phenylhydrazones.
Monosaccharides also react with phenylhydrazine (3 molar equivalents) to
yield a phenylosazone.
H H
C O + 3 C6H5NHNH 2 C NNHC 6 H5 + C6H5NH 2 + NH3 + H2O
CHOH phenylhydrazine C NNHC 6H5
an aldose phenylosazone
(an osazone)
Note that two pheylhydrazone groups are present at C-1 andC-2 in the
phenylosazone. The third molecule of phenylhydrazine is used to oxidise
the -CHOH group to C=O group and gets converted to aniline and
ammonia which are present in the products.
Osazones being solids, could be easily isolated and purified. Their

crystalline forms helped in their identification also.
You can also see that in a phenylosazone,

H H
C O C NNHC 6H5
3 C6H5NHNH2
(CHOH)n C NNHC 6H5
CH2OH (CHOH)n
unaffected portion
aldose CH2OH
phenylosazone
The crystals of the

C-2 carbon is no more a chiral centre. Hence, irrespective of the starting
osazones are very aldose having a different configuration at C-2, same phenylosazone
characteristic e.g. should result provided the configuration at other carbon atoms is the
osazone of maltose same.
forms sunflower
shaped crystals while Thus, D-(+)-glucose and D-(+)-mannose which differ in configuration only
that of lactose forms
has powder puff- at C-2 result in the same phenylosazone as shown below:
shaped. H H H
1 1 1
The crystals of the C O C NNHC 6H5 C O
osazones of
2 2 2
galactose are H C OH C NNHC 6H5 HO C H
rhombic-plate shaped
3 3 3
while those of HO C H 3 C6H5NHNH2 HO C H 3 C6H5NHNH2 HO C H
osazone of glucose 4
4 4
are needles arranged H C OH H C OH H C OH
in a broom stick 5
5 5
shape. H C OH H C OH H C OH
6 6 6
CH2OH CH2OH CH2OH
D-(+)-glucose phenylosazone D-(+)-mannose

46 (same osazone)
This reaction of D-(+)-glucose and D-(+)-mannose leading to the same

phenylosazone was studied by Emil Fischer who then concluded that Many glycosides occur
these two monosaccharides have same configuration at C-3, C-4 and C-5 naturally e.g.
anthocyanins, saponins,
carbon atoms; and they differ in their configuration at C-2 carbon atom vanillin etc.
only. Hence, D-(+)-glucose and D-(+)-mannose are epimers.
The name pyranose is
7. Formation of Glycosides derived from pyran, a
six-membered cyclic
Monosaccharides may exist in open chain form as well as in cyclic ether.
pyranose or furanose form. The cyclic form arises from intramolecular
addition of alcoholic group to the aldehyde or keto group of an aldose or a
ketose.This has been discussed in detail later. O
pyran
You know that an anomeric -OH group is present in the cyclic form of
Similarly, furanose is
monosaccharides. This can react with alcohols and form derivatives which derived from furan
are called glycosides. which is a five-
H membered cyclic ether.
6
4CH 2OH . ..
O.
HO
5 H
H 1 OCH 3 + H2O
HO 2
3 OH O
H H furan
H6
. .. methyl D-glucoside
4 CH 2OH O .
HO + CH 3OH As shownhere, D-(+)-
5 H
H methanol
HO 2
1 glucose forms two
3 OH glycosides on treatment
H OH
H
with CH3OH and HCl
6 gas.
D-glucopyranose 4 CH 2OH . ...
O
(or anomer) HO
5 H
H 1 H + H2O
HO A glycose having six
3 2
OH
H OCH 3 membered ring is known
as pyranose while its
methyl D-glucoside glycoside is called
pyranoside. Similarly, a
The formation of the glycoside linkage converts a hemiacetal of the glycose with a five
starting monosaccharide to an acetal in the glycoside. membered ring is known
as furnanose while its
You have studied in earlier course (BCHCT-133)that in an acetal,two - glycoside is called
furanoside.
OR(or-OR’) groups are linked to a carbon atom.
O OH OR" Glycosides formed by
+ ROH (alcohol) + R"OH (alcohol) glucose are called
R C H C OR' H C OR' + H 2O glucosides.
H
H H
aldehyde hemi-acetal acetal In glycosides, as the
anomeric -OH group is
Similarly, a hemi-ketal and ketal is formed if the starting material is a replaced by -OR group;
they are called O-
ketone.
glycosides. However,
O OH OR" if the -OH group is
+ ROH (alcohol) + R"OH (alcohol) substituted by -NR’R”,
R C R C OR' R C OR' + H 2O
R the glycoside is called
R R N-glycoside. Similarly,
ketone hemiketal ketal in C-glycosides, -OH
group is replaced by -
Remember that a keto group is present in fructose which is numbered as C-2 CRR’R”.
and it is this C-2 carbon which is the anomeric carbon in the cyclic form of
fructose. 47
1
CH2OH
An example of an N-
2 H
glycoside is uridine C O OH
whose structure is 6
OH
CH2OH .. OH
given below: 3 H H O CH2OH
..
O
..
NH2 HO C H 2
..
5 H OH OH C
H R''O R
N
H
4
C OH H 4 3 CH2OH HO R'
6 1
CH2OH N O OH H OH H
O Hemi-ketal
..
..
5
2 -N-glycosidic bond
H H 5  -D-fructopyranose
H 4 3 CH2OH
anomeric carbon H C OH Haworth projection
1
OH OH
 -D-fructofuranose
uridine 6
CH2OH
Fischer projection
D-fructose
Glycosides formed
from fructose are
called fructosides. In a glycoside, the carbohydrate part is called glycon while the
non-carbohydrate part is known as aglycon. Thus, in methyl β- D-glucoside
referred above, the glycon and aglycon parts can be shown as follows:
Glycosides are H
..
acetals; hence, they CH 2OH O
..
are stable in basic HO H
solutions. While in H OCH3
HO
acidic solutions, they OH aglycon
hydrolyse and H H
produce a sugar and glycon
The formation of the methyl -D-glucoside

acetal linkage by
substitution leads to Instead of CH3OH, the aglycon part can come from other alcohols, glycerol,
loss of properties
phenol, steroid etc.
such as mutarotation,
reduction etc. H
associated with the
CH2OH ..
hydroxyl group O
..
present at the HO H
anomeric carbon. HO H
R
OH O
H
aglycon
glycon
If the aglycon part is formed another monosaccharide, then the formation of

glyosidic bond between two monosaccharides yields a disaccharide.You will
study about disaccharides in detail in the next unit, i.e. Unit 16.
CH2OH CH2OH
:
H O O H
:
: H H
H H
:
OH H .. OH H
O
..
OH OH
H OH H OH
4-(-D-glucopyanosyl)-D-glucose
maltose
Since more than one -OH group is present in the monosaccharide acting as the
aglycon, the linkages resulting from these hydroxyl groups lead to different
48 isomeric compounds.
These glycosides played an important role in the determination of the

structure and (relative) configurations of monosaccharides. They were For removing the
benzyl groups to get
further used for various reactions for example, the -OH groups of the
back the -OH groups,
glycoside could be converted to benzyl, methyl, silyl ethers etc.
hydrogenolysis using
H 6 6 palladium catalyst is
CH 2OH . .. H CH OCH C H
5 O. 2 2. . 6 5
. used.
HO 4 H 5 O.
C6H5CH 2O 4 H
H OCH 3 C6H5CH 2Br
HO 2 H OCH 3
3 1 NaH, DMF,  C6H5CH 2O 2
OH H 3 H 5 C6H2 CO 1
H H H
methyl -D-glucoside
H 6CH OCH
2 3. ..
5 O.
(CH 3)2SO4 ( in excess) H3CO 4
H H
NaOH H3CO OCH 3
2
3 CH O 1
H
3 H
methyl -2,3,4,6-tetra-O-methyl-D-glucoside
pentamethyl derivative
When such a pentamethyl derivative is hydrolysed in aqueous acidic

medium, the acetal linkage breaks to give back -OH group; however, the
rest of the other -OCH3 groups remain unaffected.
1
CHO
H 6CH OCH H 6CH OCH H 2

OCH 3
2 3. .. 4 2 3. ..
4 O.
5 O. +
H3CO 5 CH 3O H
H3CO H3O H 3
H H H 1 OH
1 OCH 3 H3CO
H3CO 2 2 H 4 OCH 3
3 3 CH 3O
CH 3O H H H
H H OH
5
methyl -2,3,4,6-tetra-O-methyl-D-glucoside  -2,3,4,6 - tetra -O -methyl -D-glucose
CH 2OCH 3
pentamethyl derivative 6
The resulting hemiacetal or the cyclic form of the tetra-O-methyl derivative

is in equilibrium with its open chain form. This reaction was the basis of
structure elucidation indicating that the C-5 hydroxyl of D-glucose was
involved in the formation of the hemiacetal pyranose form.
8. Formation of Esters
One more important reaction used in the structure elucidation of

monosaccharides is their conversion to acetate esters. All the hydroxyl
groups of the monosaccharide get converted to the acetate groups on
treatment with excess of ethanoic (acetic) anhydride in a weak base.
H 6 6
CH2OH . .. H CH2OCOCH3
5 O. . ..
HO 4 (CH3CO)2O (in excess)
H 5 O.
H OH CH3OCO 4 H
HO 2 pyridine H
3 1 OCOCH3
OH H CH3OCO 2
H 3 CH OCO 1
H 3 H
-D-glucose
penta-O-acetyl--D-glucose
The formation of pentacetate of glucose indicated that it contained five

hydroxyl groups when its structure was being elucidated. 49
After understanding the reactions exhibited by monosaccharides, let us study

the structure of glucose and fructose in detail.
It is worthwhile to attempt the following SAQ at this stage.
SAQ 2
Write the product formed by the oxidation of D-glucose with dil. HNO3.
15.4 STRUCTURE OF GLUCOSE AND

FRUCTOSE
As given above in Figs.15.1and 15.2, the open chain structures of
D-(+)-glucose and of D-(-)-fructose are as follows:
1
H 1 O CH2OH
2 2 O
H OH
3
HO 3 H HO H
4
H 4 OH H OH
5 5
H OH H OH
6 6
CH2OH CH2OH
D-(+)-glucose D-(-)-fructose
You are aware from sub-Sec. 14.4.1, Sec. 14.4, Unit 14, Block 3 of
BCHCT-133 Course that aldehydes and ketones form hemiacetals or hemiketals
with alcohols. Since sugars are hydroxycarbonyl compounds, they are capable
of forming intramolecular cyclic hemiacetalsand hemiketals.
While in principle, any one of the hydroxyl group could add to the carbonyl
function; the formation of a six-membered ring is preferred, although five-
membered rings are also formed.
This is shown below in case of glucose.
CH 2OH CH 2OH
H
CH 2OH C O C O
HO C H O H H H H
H H H
C C C C O C C
OH OH H OH H OH
H OH H HO HO
C C C C C C new stereocentre
H OH H OH H OH
D-(+)-glucofuranose D-(+)-glucose D-(+)-glucopyranose

less stable more stable
Cyclic Hemiacetal Formation by Glucose
Similar cyclic structures are possible for fructose also. You will study about
50 them in the next section.
15.5 CONFIGURATION OF MONOSACCHARIDES

The configuration of monosaccharides refers to the spatial arrangement of
different groups present in them. Let us now explore about this with the
examples of glucose and fructose in more detail.
15.5.1 Absolute Configuration of Glucose and Fructose

The absolute configurations are described by assigning the configuration to
each stereocentre in the molecule. For example, D-(+)-glucose is
You may recall that
(2R, 3S, 4R, 5R)-2,3,4,5,6-pentahydroxyhexanal. R, S system of
assigning
You can see that the formation of a hemiacetal (or a hemiketal) turns the configurations was
carbonyl carbon into a new stereocentre. This leads to two new compounds discussed in Unit 11,
which are diastereomers having different configurations at C-1. Such isomers Block-3 of
BCHCT-131Course.
are called anomersand the hemiacetal or hemiketal carbon (C-1) is called the
anomeric carbon. The two anomers are differentiated by the Greek letters
and. Thus, we can call the anomers of glucose as-D-(+)-Glucopyranose
and-D-(+)-Glucopyranose. These anomers are represented below in the
modified Fischer projections using elongated lines to indicate the new bonds
formed.
O S R
C H
H C OH HO C H
H OH H OH H OH
Note that in -anomer
HO H O O
cyclisation HO H the OH at C-1 is
HO H
H OH written to the left and
H OH H OH that in the -anomer
H OH is written to right.
H H
CH 2OH
CH 2OH CH 2OH
-D-(+)-glucopyranose -D-(+)-glucopyranose
anomers
Since these modified Fischer projections do not give the correct picture of the
molecule in terms of bond lengths, Haworth introduced an alternate projection
formula called Haworth projections.
In Haworth projections, the cyclic ether is written as a planer pentagon or a

hexagon having the anomeric carbon on the right and the ether oxygen at the
top. The substituents located above and below the ring are joined by vertical Norman Haworth
lines to the ring carbons. (19 March 1883-
19 March 1950)
Note that in Haworth projections for the pyranose form in standard orientation,
the C-1 which is the hemiacetal carbon, is written in the right side and the ring The English Chemist
who received the
oxygen is written in the upper right corner. The numbering is then done in the
Nobel Prize in 1937
clockwise manner.
for his work in
6
C
carbohydrate
chemistry.
:
O
:
O 5
:
:
4 1
3 2
pyranose ring
51
Similarly, for the furanose structure, the hemiacetalcarbon is placed at the right
Glucose exists mainly side top position and the oxygen is placed at the centre at the top most position
in the pyranose form
as shown below along with the numbering of carbon atoms.
whereas fructose
forms an equilibrium 6 1
mixture of pyranose C C
: :
: :
O O
and furanose forms in 5 2
the ratio 70:30.
4 3
furanose ring
In aqueous solutions
of monosaccharides,
the pyranose form While showing the glycosidic linkages, these rings can be rotated suitably. Let
dominates, but in
biomolecules, the
us write the Haworth projections forα-D-glucopyranose and β-D-glucofuranose
furanose from occurs from their Fischer projections.
commonly.
For α–D-glucopyranose, the substituents on the right hand side in the Fischer
projection are shown down in the Haworth projection while the -CH2OH bearing
carbon, i.e. carbon number 6, is placed at the up position linked to C-5. Also
note that in α-form, the -OH group at C-1 is at the right side in the Fischer
In α-D-anomer, -OH projection.
group on C-1 is on
the right in the 6
1 CH 2OH
Fischer projection H OH
and it is shown down
:
2 H 5 O H
H OH
:
in the cyclic structure H
3 4
OH H 1
in Haworth projection HO H
and reverse is true for 4 OH 3 2 OH
H OH
β-D-anomer. H OH
5
H O
6
CH 2OH
Fischer projection Haworth projection
-D-glucose
Similarly, for β-D-glucofuranose, we first draw the Fischer projection of

D-glucose. The five membered ring containing the acetal linkage is formed by -
OH group at the C-4 carbon atom. Since the form is β, the -OH group at C-1 is
at the left side in the Fischer projection.
H 1 O 1
C HO H
2 2
H OH H OH
3 3
HO H HO H
4 4
H OH H O
5 5
H OH H OH
6 6
CH 2OH CH 2OH
D-glucose
Fischer Projection
Again, the Haworth projection is written by drawing the furanose ring and
placing the right hand side substituents of the Fischer Projection on the down
52 side in the Haworth projection.
6
CH 2OH
: :
HO 5C HO OH
4 OH H 1
H 3 2
H
H OH
-D-glucofuranose
Haworth projection
The envelope conformation of the furanose form can be represented as shown

below.
CH 2OH
CHOH O
H
H
OH
HO OH
H
H
-D-glucofuranose
Earlier, in 1909, according to an American Chemist C.S. Hudson, in

D-sugars, the most dextrorotatory of α/β anomers is called α and the other one
is named as β. Similarly, for L-sugars, the more levorotatory anomer is named
as α-anomer while the lesser levorotatory anomer is known as β-anomer.
But later, a system based on absolute configuration has been adopted wherein
the anomer having the hemiacetal hydroxyl on the same side as the hydroxyl
group reacting with the aldehyde or keto group (cis) is called α-anomer while
the anomer in which the hemiacetal hydroxyl is on opposite side, i.e. trans is
called β-anomer.
From Haworth projections of D-(+)-Glucose, we can write its conformations.

Since its pyranose form is a six-membered ring in which C-O-C bond angle is
IIIo, its chair conformations can be written similar to the cyclohexane molecule.
Now two alternate chair forms are possible, e.g. for β-D-(+)-glucopyranose. We
can write them as follows:
6
H 6 CH2OH OH
: :
4 CH2OH O 5 1
5
: :
H O
HO H OH H
H 1 OH H
HO 2 H
OH 4 3 2
3 H
H H OH OH
(I) (II)
(64%) (minor)
(C1) (1C)
4
C1 conformation 1
C4 conformation
-D-(+)-glucopyranose
In structure (I), carbon number 1 is down whereas carbon number 4 is up, so it

is written as 4C1 conformation and is called C1 conformation. However, in
structure (II), C-1 carbon is up and C-4 carbon is down; therefore, it is written
as 1C4 conformation and is also called 1C conformation.
Note that C-6 carbon atom bearing the primary hydroxyl group (-CH2OH) is
axial or up in the 1C conformation structure (II) whereas it is equatorial in C1
conformation structure (I). 53
You can also see in structure (I) that all bulky –OH groups are in equatorial
positions; therefore, this conformation is lower in energy and hence,more
stable due to lower interactions between the bulky groups as compared to the
conformation shown in structure (II).
As a result, the C1 conformation shown above in structure (I), predominates

and occurs to the extent of 64% in the aqueous solution of D-Glucopyranose;
however, the conformation shown in structure (II) is present in the minor form.
The other conformer which is present to the extent of 36% is α-D-

glucopyranose in 4C1 conformation as shown in structure (III) below while its
1
C4 conformation, structure (IV), is present in minor amount in the solution.
6
H CH 2OH H
6
CH2OH 1
: :
O 5
: :
4 5 H H OH O OH
HO
H H H H
HO 2 3
3 OH 1 4 2
H
H OH OH OH
(III) (IV)
(36%) (minor)
4 1
C1 conformation -D-(+)-glucopyranose C4 conformation
You can see in structure (III) that -CH2OH group is occupying the equatorial
position alongwith -OH groups at C-2,C-3 and C-4 carbon atoms. Remember
that the -OH group at C-1 has to be axial as it is the α-D-(+)-glucose.
An interesting fact about β-D-(+)-glucoseis that amongst all D-aldohexoses,

only this is the one which can have a conformation shown by structure (I) in
which all bulky groups occupy equatorial positions. Hence, it is the most
abundant form in nature.
The formation of α-anomer can be explained on the basis of anomeric effect.

The lone pair -lone pair interactions are unfavourable between the oxygen of
anomeric -OH and the ring oxygen atom in the β-anomer whereas these
interations or repulsions are reduced when the -OH group occupies the axial
position in α-anomer.
6
unfavourable interaction
H :
:
CH2OH H 6
:
:
O CH2OH
4 5 favourable interaction :
:
HO H O
4 5
H 2 H HO H OH
HO H
3 OH 1 HO 2 1
H : OH 3 OH H
H
:
-anomer  -anomer
Mutarotation is
catalysed by acids The anomeric effect is more prominent in non-polar solvents and decreases
and bases. with the inecrase in the polarity (dielectric constant) of the solvent such as
In Latin, mutare
means to change. water.
15.5.2 Mutarotation
Both -D-(+)-glucopyranoseand -D-(+)-glucopyranose are optically active but
differ widely in their optical rotations. The -D-(+)-glucopyranose has the
54 specific rotation of [α ]D298 Κ  112ο whereas -D-(+)-glucopyranose has
ο
18.7 - When dissolved in water, the optical rotation of the solution
298 Κ
[α ]D 
gradually changes with time until it reaches an equilibrium value of +52.7°.

This is because -D-(+)-glucopyranose rapidly establishes an equilibrium with
a small amount of the open-chain aldehyde form which in turn undergoes
renewed and reversible ring closure to the -anomer.
1
CHO H 6 ..
H 6 .. HOCH2 O
..
HOCH2 O
.. H 2 OH 4
4 HO 5 H
HO 5 H H OH
H H 3
+ 
H or HO HO H +
H or HO

HO 2 1
HO 2 1 3 OH H
3 OH HO 4
H OH H
H -D-(+)-glucopyranose
-D-(+)-glucopyranose 5
H OH 298 K
298 K
o []D = + 18.7o
[]D = + 112 6
CH 2OH (63.6%)
(36.4%)
aldehyde form
(0.003%)
This interconversion is called mutarotationand was first observed in 1846. It

involves change of configuration at one stereocentre in a compound having
more than one such centres.
SAQ 3
What is the relationship between α-D-(+)-Glucose and β-D-(+)-Glucose?
SAQ 4
What are anomers?
15.6 ASCENDING AND DESCENDING OF CHAINS

IN MONOSACCHARIDES
We will focus our attention on the methods which are used to convert one
aldose to another aldose which has either one more carbon atom in the chain
or one less carbon atom in the chain. The former conversion is called
ascending or lengthening of carbon chain whereas the latter is known as
descending of carbon chain. Let us first study the method for ascending the
carbon chain.
Kiliani-Fischer Synthesis
Kiliani showed that cyanohydrins obtained from aldoses could be hydrolysed

to aldonic acids.
new stereocentre
CN CN
CH O
H OH HO H
HO H
HCN HO H + HO H
H OH Heinrich Kiliani
H OH H OH
H OH University of Freiburg
H OH H OH
CH 2OH (30th October 1855-
CH 2OH CH 2OH 25th February 1945)
D-arabinose D-gluconitrile D-mannonitrile
(I) (29%) (51%)
(IIa) (IIb) 55
In this method, an aldose is treated with hydrogen cyanide to give the
cyanohydrins. Let us understand this with the example of D-arabinose (I).
Note here that the formation of the cyanohydrins has led to the generation of a
new stereocentre; therefore, the product formed is a mixture of diastereomers
(IIa and IIb) which are obtained in different amounts. These two diastereomers
are epimers and they differ in configuration at C-2 carbon.
The hydrolysis of the above mixture of cyanohydrins yields a mixture of

aldonic acids (IIIa and IIIb) as given below.
COOH COOH
H OH H OH
The mixture of aldonic
HO H HO H
acids is separated at +
this stage. H OH H OH
H OH H OH
CH 2OH CH 2OH
IIIa IIIb
aldonic acids
Fischer developed a method for the conversion of aldonic acids to aldoses by

the formation of lactones which were reduced to the aldose.
COOH COOH
H OH HO H
HO H HO H
+
H OH H OH
H OH H OH
CH 2OH CH 2OH
III a III b
_ aldonic acids
The OH groups
(diastereomers)
present at and 
positions can form H2O H2O
lactones. The 
lactone is formed is
more stable. C O C O
H OH O HO H O
HO H HO H
H H
H OH H OH
CH 2OH CH 2OH
IV a IV b
aldonolactones
(diastereomers)
Na(Hg), Na(Hg),
CO2 reduction reduction
CO2
CHO CHO
H OH HO H
HO H HO H
H OH H OH
H OH H OH
CH 2OH CH 2OH
Va Vb
aldohexoses
56 (diastereomers)
The cyanohydrins obtained (IIa and IIb) in Kiliani-Fischer synthesis outlined

above could be separated. As an alternative these on catalytic reduction in
aqueous acid yield the aldoses which have one more carbon atom in their
chain as compared to the starting aldose.
CN CN
H OH HO H
HO H HO H
H OH H OH
H OH H OH
CH 2OH CH 2OH
IIa II b
H2, Pd-BaSO4 H2, Pd-BaSO4
HC NH
HC NH
H OH HO H
HO H HO H
H OH H OH
H OH H OH
CH 2OH CH 2OH
VI a VI b
H+, H2O, H+, H2O,
NH3 NH3
CHO CHO
H OH HO H
HO H HO H
H OH H OH
H OH H OH
CH 2OH CH 2OH
Va Vb
After understanding how to increase the chain length of an aldose by one

carbon atom, let us study how we can decrease the chain length of an aldose
by one carbon atom.
Prof. Otto Ruff
One method used for this purpose is Ruff degradation. It involves oxidative
(12 December, 1871-17th
th
decarboxylation. The aldose whose length is to be shortened is first oxidised September, 1939)
to the aldonic acid by bromine water. The calcium salt of the aldonic acid is University of Danzig,
Germany
oxidised by hydrogen peroxide in the presence of ferric salt which gives the
desired aldose and the carbonate ion. 57
Let us understand these steps by the example of conversion of an aldohexose

to an aldopentose.
- 2+
CHO COOH COO ) 2 Ca
H OH H OH H OH
HO H Br2, H2O HO H CaCO3 HO H
H OH H OH H OH
H OH H OH H OH
CH 2OH CH 2OH CH 2OH
an aldohexose an aldonic acid calcium aldonate
Fe3+, H2O2
CHO
HO H
2-
H OH + CO 3
H OH
CH 2OH
an aldopentose
SAQ 5
What products will be obtained by Kiliani-Fischer synthesis of
D-(+)-glyceraldehyde?
15.7 SUMMARY
 Carbohydrates constitute an important class of compounds which have
diverse functions.
 Some common examples of carbohydrates are cellulose which gives

structural support to plants, starch which serves as the reserved energy
source and sugars such as sucrose and glucose. Glucose is an essential
constituent of blood in higher animals.
 Monosaccharides are the simplest carbohydrates which cannot be

hydrolysed into smaller and simpler carbohydrates.
 A monosaccharide may be further classified as an aldose or a ketose if it

contains an aldehyde or a keto group.
 Depending upon the number of carbon atoms in the chain, sugars are
called trioses(3 carbons), tetroses(4 carbons), pentoses (5 carbons),
hexoses(6 carbons), and so on.
 Monosaccharides undergo a variety of reactions such as oxidation, reduction

58 and formation of osazones and glycosides.
 Formation of hemiacetal (or hemiketal) turns the carbonyl carbon into a

new stereocentre. This leads to twonew compounds which are
diastereomers having different configurations at C-1. Such isomers are
called anomers and the hemiacetal or hemiketal carbon (C-1) is called
the anomeric carbon.
 The structures of monosaccharides can be represented by Fischer

projections and Haworth projections.
 Mutarotation involves change of configuration at one stereocentre in a

compound having more than one such centres.
 In monosaccharides, ascending of carbon chain can be done byKiliani –

Fischer synthesis and descending of carbon chain can be done by Ruff
degradation.

1. Draw the Haworth projection for D-(+)-xylose and D-(+)-glucose.
2. Write the Fischer projection of D-(+)-xylose and D-(+)-xylulose.
3. Write the product formed by the oxidation of D-(+)-glucose with bromine

water.
4. Which compound will form same phenylosazone as D-(+)-galactose?
5. Write the Haworth projection of α-D-fructofuranose.
6. Draw the possible chair conformations of β-D-(+)-glucose. Which one of

these conformations is more stable and why?
7. Name the product formed by Ruff degradation of D-(+)-glucose. Which

other aldose will give the same product on Ruff degradation?
15.9 ANSWERS
1. i) Ribose is a pentose while fructose is a hexose.
ii) Xylulose is a ketose having a five carbon chain.
iii) D-erythrose is an aldose having four carbon atoms in the chain.
2. CHO COOH
H OH H OH
HO H HO H
H OH HNO 3 H OH
H OH H OH
CH2OH COOH
D-glucose D-glucaric acid
3. They are diastereomers or anomers.

59
4. The diastereomeric pair of monosaccharides which differ in configuration

at C-1 carbon atom are called anomers.
5. D-(+)-erythrose and D-(+)-threose.
Terminal Questions
1. H CH2OH
5 .. 5 ..
H .O. H H .O
. H
H 1 H
4 4 1
OH H OH H
HO OH HO OH
3 2 3 2
H OH H OH
D-(+)-xylose D-(+)-glucose
2. H 1 O 1
CH 2OH
C
2
2 O
H OH
3
3
HO H
HO H
4
4 H OH
H OH 5
5 CH 2OH
CH 2OH
D-(+)-xylose D-(+)-xylulose
3. CHO COOH
H OH H OH
HO H Br2 HO H
H OH H2O H OH
H OH H OH
CH2OH CH 2OH
D-(+)-glucose D-(+)-gluconic acid
4. D-(+)-talose
5. 6 1
: :
HOH 2C O CH2OH
5 H OH 2
H 4 3 OH
OH H
-D-fructofuranose
6
CH 2OH OH
6. H 6
5
: :
4 CH2OH O 1
5
: :
H H OH O H
HO
H 1 OH H H
HO 2 3
OH 4 H 2
3
H H OH OH
equatorial bulky groups axial bulkyl groups
The first conformation is more stable because all bulky substituents are
equatorial.
7. D-(+)-arabinose. D-(+)-mannose will also give the same pentose.
60
Unit 16 Carbohydrates - II: Dissaccharides and Polysaccharides
UNIT 16
CARBOHYDRATES-II:
DISACCHARIDES AND
POLYSACCHARIDES
Structure
16.1 Introduction 16.3 Polysaccharides
Expected Learning Outcomes Starch
16.2 Disaccharides Cellulose
Maltose 16.4 Summary
Cellobiose 16.5 Terminal Questions
Lactose 16.6 Answers
Sucrose
16.1 INTRODUCTION
In Unit 15 on monosaccharides, you have studied about the basic aspects of
carbohydrates and monosaccharides. Therein, we have discussed the
important monosaccharides i.e. glucose and fructose in detail. Having
understood the basic aspects of monosaccharides and how to write Fischer
and Haworth projections of monosaccharides, let us now widen our window to
the study of other important classes of carbohydrates, i.e., disaccharides and
polysaccharides.
You already know that disaccharides contain two monosaccharides linked to

each other. In this unit, we will be discussing about some important
disaccharides such as maltose, cellobiose, lactose and sucrose. In addition to
these, we will also elaborate on the structure and importance of starch and
cellulose which are polysaccharides containing many glucose monosaccharide
units linked to each other.

able to:
 define disaccharides and polysaccharides;

61
 give examples and importance of some common disaccharides and

polysaccharides;
 explain glyosidic linkage;
 write the structures of disaccharides such as maltose, cellobiose, lactose

and sucrose;
 name the above disaccharides and discuss their important;
 properties; and
 describe the structural features and importance of polysaccharides such

as starch and cellulose.
16.2 DISACCHARIDES
You already know that disaccharides are the carbohydrates formed by the
linkage of two monosaccharide units and on hydrolysis they yield two
molecules of monosaccharides. These two monosaccharide units can be
same or different, as you will study in case of different disaccharides
discussed in this section.
The structure, properties and biological roles of carbohydrates can be easily

understood by their three dimensional shapes. Hence, we will be emphasising
on the type of linkages joining the monosaccharides.
Let us now refresh some of the concepts you have learnt in Unit 15. This will
help you to understand the forthcoming discussion on writing of structures and
nomenclature of disaccharides. Now, carefully revise the following points:
 An anomeric –OH group is present in the cyclic form of monosaccharides.

This can react with alcohols and form derivatives which are called
glycosides.
H
6
4CH 2OH . ..
O.
HO
5 H
H 1 OCH 3 + H2O
HO 2
3 OH
H H
H6
. methyl D-glucoside
4 CH 2OH O...
HO + CH 3OH
5 H
H methanol
HO 2 1
3 OH
H OH
H
6
D-glucopyranose 4 CH 2OH . ...
O
(or anomer) HO
5 H
H 1 H + H2O
HO
3 2
OH
H OCH 3
methyl D-glucoside
 In a glycoside, the carbohydrate part is called glycon while the

non-carbohydrate part is known as aglycon. Thus, in methyl D-glucosides
(α-and β-) referred above, the glycon parts are shown in the black colour
62 while the aglycon parts are indicated by the red colour.
 The formation of the glycoside linkage converts a hemiacetal of the

starting monosaccharide to an acetal in the glycoside. Remember that in
an acetal two –OR (or –OR’) groups are linked to a carbon atom.
 Glycosides are acetals; hence, they are stable in basic solutions. While in
acidic solutions, they hydrolyse and produce a sugar and the alcohol.
 A glycose having six membered ring is known as pyranose while its

glycoside is called pyranoside. Similarly, a glycose with a five membered
ring is known as furanose while its glycoside is called furanoside.
 Glycosides formed by glucose are called glucosides.
 Instead of CH3OH, aglycon part can be derived from other alcohols,

glycerol, phenols, steroid etc. If the aglycon part is another
monosaccharide, then the formation of glyosidic bond between two
monosaccharides yields a disaccharide.
 Thus, the monosaccharides can link to each other through the glycosidic
linkage.
Having refreshed the above points, let us also understand what is meant by
the term reducing and non-reducing sugars before proceeding to the study of
disaccharides.
Reducing and Non-reducing Sugars
Carbohydrates which contain the hemiacetal group give positive Tollens, The formation of the
Fehling and Benedicts tests and hence, these are called reducing sugars. acetal linkage by
substitution leads to
This is so because in an aqueous solution, there is an equilibrium between
loss of properties
the hemiacetal and the open chain structure of the sugar. It is the open chain such as mutarotation,
form which gets oxidised and the equilibrium keeps on shifting to give more of reduction etc.
open chain form. associated with the
hydroxyl group.
However, formation of an acetal linkage does not allow such equilibrium and
those sugars i.e. glycosides which contain acetal linkages instead of
hemiacetal linkages, do not give positive tests with Tollens’ reagent, Fehling
reagent and Benedict’s reagent and are called non-reducing sugars.
You will study more about reducing and non-reducing nature of sugars when
you study the structures of disaccharides.
Having refreshed the above knowledge, let us know focus our attention on
some very interesting disaccharides which are very commonly used.
16.2.1 Maltose
The hydrolysis of starch using the enzyme diastase yields maltose as one of
the products. It has molecular formula C12H22O11 and is dextrorotatory in
nature. Some of its properties are listed below:
 One mole of maltose on acid-catalysed hydrolysis or hydrolysis using the

enzyme maltase (from yeast) gives two moles of D-(+)-glucose. 63
Maltase hydrolyses only α-glycosidic linkages and not β-ones. Therefore,

in (+)-maltose, two D-(+)-glucose units are linked to each other by
α-glycoside link in which the α-anomeric –OH of one glucose unit is linked
to the –OH group of the other glucose unit.
 Maltose reduces Tollens reagent and Fehling’s solution and is called a

reducing sugar. This indicates that one monosaccharide unit in the
glucoside exists in the hemiacetal form. The presence of one hemiacetal
is also indicated by the bromine water oxidation of maltose to maltonic
acid, which is a monocarboxylic acid.
Note that the details  The methylation of maltonic acid followed by hydrolysis and methylation of
of these reactions are maltose followed by hydrolyses indicated that the –OH group at C-4 of one
beyond the scope of glucose unit is involved in the formation of glycosidic linkage and the
this course and you reducing hemiacetal linkage is present in the pyranose ring.
will study them at a
higher level. All these facts led to the following structure of maltose. Please also
understand the numbering of the carbon atoms shown below:
anomeric
carbon
6 6
CH2OH
. .. CH 2OH. .
5
O .H 5 .
H H O .H +
4 H 1 + 4
H 1 H
OH H OH H
HO OH HO OH
3 2 3 2
H OH H OH
-D-(+)-glucose -D-(+)-glucose
(monosaccharides)
6 6
CH2OH CH 2OH
5 . .. 5 . ..
H O. O.H
1 H H
4 H H 1 + H 2O
OH H O 4 OH H
HO OH
3 2 3 2
H OH H OH
orientation of this -OH

is down which indicates the
The type of linkages  linkage
present here are
-maltose
same as those (a disaccharide)
present in
amylopectin, about The –OH group at C-1 can be represented by a wavy line as shown below
which you will study
which indicates that it can take either α- or β- orientation.
in the later section i.e.
Sec.16.3.
H
CH2OH
HO ...
H H O.
HO
H
HO CH2OH
H ... ...
O. O.
H H
HO
HO
H OH
64
Thus, according to the position of the –OH group we get two anomeric forms
i.e., α-anomer and β-anomer. The specific rotations of these α-(+) and
β-(+) forms being, +168o and +112o at 25o C, respectively.
The structure of β-anomer of maltose is given below along with its name:
6
6
CH2OH CH 2OH hemiacetal
5 . .. acetal 5 . ..
OH
H O .H H O.
H 1 H 1
4 .. 4 OH
OH H H
HO O.. H
3 2 3 2
H OH H OH
4-O-( -glucopyranosyl)-  -D-glucopyranose
maltose
H 6
CH 2OH . ..
5 O.
HO 4  linkage
H  anomer of glucose
H 1 H
HO 2 H 6 (the agylcon)
3 OH . . CH 2OH . ..
H 5 O.
.O. 4
H
H OH
HO 2 1
(1,4)-glycosidic bond 3 OH
H H
4-O-( -glucopyranosyl)-  -D-glucopyranose
maltose
For understanding how to arrive at the names of disaccharides, let us study

the recommendations given below. But before that you can answer the
following SAQs.
SAQ 1
Does maltose show mutarotation?
SAQ 2
Write the structure of maltonic acid.
Nomenclature of Disaccharides:
As per the Recommendations (1996) of Joint Commission of

IUPAC and International Union of Biochemistry and Molecular
Biology, a disaccharide in which a free hemiacetal is present can
be named as a glycosylglycose. The locants of the glycosidic
linkage and the anomeric descriptor being mentioned in the name.
The locants are given in brackets between the glycosyl and glycose
units in one way of nomenclature related to oligosaccharides. The
other way is to give the locants before (in front of) the glycosyl
prefix, similar to names given to glycosides as discussed earlier.
Hence, according to these two ways, maltose can be named as:
i) α-D-glucopyranosyl-(1→4)-β-D-glucopyranose
[α-D-Glcp (1→4)-β-D-Glcp] or
65
ii) 4-O-α-D-glucopyranosyl-β-D-glucopyranose
H 6
4 CH 2OH . ..  linkage glucose unit
O.
HO 5 H
H 1 H
HO 2 H 6
3 OH 4 CH 2OH . ..
H . .. O.
O. 5 H  anomer,
H 1 OH
HO 2 as the -OH is upside
glucose unit 3 OH H
H
 glycosidic bond
The second method of nomenclature is used by Chemical Abstracts Service.

Also note that O-locant is not used in the first way of naming.
In the first way of nomenclature, you can also see that a shorthand form of
notation is given below the name in square brackets. Such a shorthand can
be easily correlated with the name given above for maltose.
You will see later that this shorthand representation is quite useful when
higher oligosaccharides (or polysaccharides) are named.
16.2.2 Cellobiose
Cellobiose is obtained by the partial hydrolysis of cellulose. It has molecular
formula C12H22O11. The cellobiose shows the properties similar to (+)-maltose
discussed above.
On acid-catalysed hydrolysis, it yields two molecules of D-(+)-glucose. It is a

reducing sugar and a sequence of reactions such as oxidation, methylation
and hydrolysis indicated that a glucoside linkage is present between the two
glucose rings in their pyranose forms, involving–OH group at C-4 carbon
atom.
It is hydrolysed by the enzyme emulsin which hydrolyses on β-glucoside

linkages; hence two glucose units are linked by a β-glucoside linkage in
(+)-cellobiose. Cellobiose is not hydrolysed by α-glucosidase enzyme maltase.
You can show the β linkage in the following two ways:

6
CH 2OH anomeric carbon;
5 .. OH is upside so  anomer
H . . OH
O
4
H
6 OH H   linkage
CH2OH . H
6 CH OH
2
5 . . O 3 5 .. 6 CH OH
.. .. H
H O 2
OH H O
. . H . . OH
H 1 H 1 5
O
4
OH H 4 OH .. 4 . .1 anomeric carbon;
H O H OH is upside so anomer
HO H -1, 4-glycosidic linkage HO H . . OH H
3 2  3 2 H
H OH H OH
H OH
4-O-(  -D-glucopyranosyl)-  -D-glucopyranose
Cellobiose also shows mutarotation; hence it can exist as α- and β-anomers.

In the above two structures, the β-anomer has been shown as the hydroxyl
group at C-1 anomeric carbon atom is up.
66
If we have to write the structure of α-anomer, we will write the anomeric –OH
at C-1carbon atom down and this is shown below:
6
CH 2OH
5 ..
H
H .. H
O OH down, - anomer
4 OH H 1
6
CH 2OH . .
5 . ..O. 3 2 OH
H O
.. H OH
H
4 OH H 1
HO
3 2 H
H OH
4-O-(  -D-glucopyranosyl)- -D-glucopyranose
Let us next study another disaccharide, lactose. But before that attempt the
Lactose in milk gets
following SAQ.
broken by the
enzyme lactase to
SAQ 3 glucose and
galactose in small
What is the name of β-anomer of (+)-cellobiose? intestine. Many
people have lactose
intolerance because
not enough lactase is
16.2.3 Lactose produced in their
bodies; the reasons
Lactose occurs in human milk, cow’s milk and milk of other mammals as well. for that may be
Its molecular formula is C12H22O11.Similar to the disaccharides studied above, different. In such
people, lactose on
it is also a reducing sugar and undergoes mutarotation. reaching to colon
mixes with bacteria
The acidic hydrolysis of (+)-lactose yields equal amounts of D-(+)-glucose and and gets fermented
D-(+)-galactose. It is also hydrolysed by emulsin enzyme which hydrolyses β- which results in
glycosidic linkages; therefore, lactose is formed by β-glycosidic linkage symptoms like gas,
diarrhoea and
between D-(+)-glucose and D-(+) galactose. bloating.
Lactose exhibited the following reactions:
 Osazone formation and its hydrolysis
 Oxidation followed by hydrolysis

The details of these
 Oxidation, methylation and hydrolysis. reactions are beyond
the syllabus and you
These reactions indicated that will study them at a
higher level.
 the two monosaccharide units - glucose and galactose exist in pyranose
forms in lactose.
 the galactose forms β-glycosidic linkage with the hydroxyl group at C-4 of
glucose unit. The anomeric –OH of the glucose, being free, exhibited
oxidation and osazone formation. The mutarotation showed by lactose
was also due to this anomeric -OH group.
67
D-(+)-glucose and D-(+)-galactose differ in their configurations at carbon

number 4 only. Thus, the structure of β-anomer of lactose can be written as
follows:
6
CH2OH
5 . . OH
6
H O..
CH2OH 4
H
OH H 1
5 .. .. H
HO O.. 1 O
.. 3 2
H
4 OH H OH
H
H
3 2
H OH
OH 6
CH2OH . . H 6
H 4 5 O
.. CH2OH . .
H .. 4 5 O
..
H 1
O
.. H
HO 2 H 1
3 HO OH
OH 2
H H 3 OH
H H
A galactoside is the  lactose
glycoside of 4O( Dgalactopyranosyl) Dglucopyranose
galactose. Refer to
naming of glycosides You can see in the above structure that a D-galactosyl unit is linked to the
again. hydroxyl group present at C-4 of the glucose. As the anomeric -OH of
galactose is linked to the -OH group at C-4 of glucose, lactose is a galactoside
not a glucoside.
SAQ 4
Write the Howarth projection for α-lactose. Also give its name.
SAQ 5
What is the other way to write the name of α-anomer of lactose?
16.2.4 (+)–Sucrose
(+)-Sucrose is the most common table sugar with which you come across
daily. This is the most widely occurring disaccharide which is obtained from
sugarcane and sugar beets.
Sucrose has molecular formula is C12H22O11 and on acid-catalysed hydrolysis

Honey contains invert its one mol yields one mol of D-(+)-glucose and one mol of
sugar where D-(+)-fructose.
honeybees
useinvertaseenzyme An interesting property of sucrose is that it itself is dextrorotatory while its
for hydrolysis. products of hydrolysis show net levorotation due to the greater value of
levorotation by D-(-)-fructose. The resulting mixture is called inverted sugar or
invert sugar.
68
hydrolysis
(+) - sucrose D- (+) - glucose + D- (-) -fructose
o
invertase enzyme
specific rotation o o
+ 66.5 +52.7 - 92.4

It does not reduce Tollens reagent and Fehling’s solution. Thus, it is a non-
reducing sugar. It does not form an osazone and does not show
mutarotation which indicates absence of free aldehyde or ketone group
which can form a hemiacetal.
Thus, the glycosidic linkage is present between the C-1 of glucose and C-2 of
fructose to give the acetal linkage.
Sucrose is hydrolysed by α-glucosidase enzyme obtained from yeast and

not by β-glucosidases. Thus, there is α-configuration at the glucosidic
(glycosidic) linkage.
Similarly, the hydrolysis by an enzyme sucrase which hydrolyses only β-

fructofuranoside linkages indicates that fructoside portion is linked by β-
glycosidic linkage to the hydroxyl group at the anomeric carbon, i.e. C-1 of
the glucose portion.
Thus, in sucrose, the glucose portion in its pyranose form is linked by α-

glycosidic linkage to fructose portion in its furanose form by β-glycosidic
linkage. Hence, the following structure emerges for sucrose.
6 1 6
CH2OH CH2OH CH2OH
mirror
5 .. .. 5 ..
H .O. H O
.. H H O
.. H D-fructose L-fructose
4
H 1 H
OH H .. 2 H HO 5 -glucose 4 OH
H
1
1 1
HO O CH2OH  - glycosidic linkage CH2OH CH2OH
3 2
.. 6 HO
2C
3 4 3 2 2C
O O
H OH OH H H HO . .
- glucosidic linkage 3
HO C H H 3C OH
6
O
.. 4 4
H C OH HO C H
CH2OH. .
O  - glycosidic linkage H 5C OH HO 5C H
..
6  - fructose5 2 6CH OH 6CH OH
H CH H HO 2 2
2OH . . H
O
1
4 3 CH2OH cy
5 .. cli
n
CH2OH 1
HO 4 ..
tio
OH H sa
H
sa
H O
.. H tio
n
cli
HO 1 H
cy
3
2 H HO 5
OH O .. 2 CH2OH 6 1
H .. 4 6 CH2OH CH2OH 6
3 .. ..
CH2OH OH
OH H O
.. mutarotation O
- glucosidic linkage ..
5 2 5
H HO H HO 2
H OH H CH2OH
sucrose 4 3 4 3 1
OH H OH H
 D - fructofuranose  D - fructofuranose
The disaccharides obtained by the formation of glycosidic linkages between

the hydroxyl groups of two anomeric groups of two monosaccharide units are
named as glycosyl glycosides, according to the recommendations of the
Also note a double
Joint Commission referred above. According to the recommendations, for headed arrow is
choosing the parent glycoside, the aldose is preferred over the ketose; placed between 2 and
therefore, sucrose is named as β-D-fructofuranosyl-α-D-glucopyranoside but 1 in the shorthand
name and think why
not as α-D-glucopyanosyl-β-D-fructofuranoside.
this is so.
Thus, the name of sucrose in shorthand notation can be written as
69
[β-D-Frucf-(2↔1)-α-D-Glcp]
Before studying the polysaccharides in the next section, it would be

worthwhile to know the nomenclature of oligosaccharides.
Nomenclature of Oligosaccharides
i) Oligosaccharides without a free hemiacetal group
Such oligosaccharides can be named as glycosylglycosyl glycoside or

glycosylglycosylglycoside,where the parent chain will be decided by rules of
nomenclature as listed in the Recommendations 2-carb-2.1.
Alternatively, an end to end sequential method is used for naming, e.g. as in

case of raffinose shown below:
6
CH2OH galactose
5 . ..  -linkage
HO O .H
4 H
OH H 1 (1 6) -O-glycosidic linkage OH 6
CH 2OH . ..
H 5 O.
3 2 H 4 H
H OH. H 1 H
.. HO 2  -linkage
O. 3 OH
.. O..
H
6 fructose
CH 2 . .  -linkage 1CH OH H 6
CH 2 . ..
H 5 O
.. H 2 .. 5 O.
H .O. H HO 4
2 H
4 OH H 1 .. H 1 H
.O. H HO HO 2
HO 5 3 OH
3 2
..
H
OH
3 4 .O. OH H 6
H OH H CH
6
2OH 3 4 CH 2OH
-linkage
glucose (1 2) -O-glycosidic bond 2 H HO 5
..
HOCH 2 O H
-D-galactopyranosyl-(1 6)--D-glucopyranosyl- -D-fructofuranoside 1 ..
ii) Oligosaccharides with a free hemiacetal group
The name of such oligosaccharides is given as glycosyl[glycosyl]n glycose.

The glycose or the reducing sugar being written on the right or down stream
portion.
H 6  - linkage
CH 2OH . .. H 6
O. CH2OH .. . H 6
HO 4 5 O. CH 2OH . .. anomeric carbon
H .. 4 5
H O H .. 5 O.
HO 1 .. H 4
2 HO 1 .O. H
H
3 OH H 2 HO 1
H 3 OH H 2
H 3 OH OH
 - linkage H
reducing sugar
 -D-glucopyranosyl-(1 4)-  -D-glucopyranosyl (1 4)-D-glucopyranose
In case of branched chains, the longest chain is regarded as the parent chain.
If two chains of equal length are possible, then the chain having lower locants
for the branch point is chosen as the parent chain.
The terms designating the branches are enclosed in square brackets. An

example to illustrate this is shown below:
70
H 6
CH2OH . ..
HO 4 5 O.
H
H 1 H
HO 2
3 OH
H 6 H
CH 2OH . . ..
5 . O
. O
. .
HO 4
H
H 1 H
HO 2 H6
3 CH 2 ..
OH .
H . 4 5 .O.
.O. H
H
HO 1
2
3 OH OH
H
-D-gulcopyranosyl-(1 4)-[ -D-glucopyranosyl-(1 6)-D-glucopyranose
(isopanose)
Ref: gmul.accuk/sbcs/iupac/2carb/37.html#373
16.3 POLYSACCHARIDES
Polysaccharides are naturally occurring polymers which contain hundreds or
thousands of monosaccharide units. The polysaccharides formed from a
single monosaccharide are called homopolysaccharides while those formed
from more than one type of monosaccharide are called heteropoly-
saccharides. There are three important polysaccharides formed from the
same monomer, i.e. glucose. These are starch, cellulose and glycogen.
These homopolysaccharides have different structures, linkages and
properties. While starch and cellulose form food reserves and structural
material, respectively in plants; glycogen is a carbohydrate reserve in animals.
Similar to the approach followed for disaccharides, here also, you will study
the structures of starch and cellulose. The knowledge of linkages present
between them will help you to understand how their properties and functions
are related to their structure.
The homopolysaccharides are also called glycans. Those ending in the name
of the monosaccharide is replaced by an ending in the polysaccharide. Thus,
starch, cellulose and glycogen are called glucans (from glucose). Similarly,
the homopolysaccharides of galactose are known as galactans. Let us now
study about starch and cellulose in detail.
SAQ 6
What is general formula for starch, cellulose and glycogen?
16.3.1 Starch
The major sources of starch are corn, potatoes, wheat, rice etc. which are
In the name, the
used as sources of carbohydrates in food by humans. It occurs in the roots,
linkages between the
tubers and seeds of plants as microscopic granules. These granules are
carbon atoms forming
insoluble in cold water but they swell in hot water, burst and release the glycosidic bonds
water-soluble fraction called amylose which constitutes about 10-20%. The precede the symbols
rest 70-80% water insoluble portions is called amylopectin. designating the
configuration of
Amylose on hydrolysis yields only (+)-maltose which further hydrolyses to sugars.
yield D-(+)-glucose. This indicates that amylose contains D-(+)-glucose units 71
linked to each other by α-glycosidic bonds; similar to the one you studied
above in case of (+)-maltose. Remember that the C-1 anomeric hydroxyl
group forms a glycosidic bond with the -OH group at C-4 of the other glucose
unit. Thus, a part structure of amylose can be represented as shown below:
H 6
CH 2OH . ..
. .. 5 O.
O. 4 H
H 1 H
HO 2
3 H 6
OH CH 2OH . ..
H . .. 5 O.
O. 4 H  - linkage
H 1 H
HO 2
 - linkage 3 H 6
ma OH CH 2OH . ..
ltos H . O .. 5 O.
e lik . 4
e st H
ruc H 1 H
ture HO 2
3 OH
H ..O..
[ 4 )-D-Glcp-(1 ] n
Amylose (part structure)
(1 4)--D-glucopyranan (as per glycan nomenclature)
Ref: Fig. ctri.in/modifiedstarches/intriomodifiedstarches.aspe
The next question then arises, how many such D-(+)-glucose units are joined
together to form a chain. The studies about physical and chemical properties
have shown that chains contain more than 1000 units to 4000 units and the
molecular weight ranges between 1,50,000-6,00,000. Such a chain is shown
below:
H 6 ..
CH 2OH .O
5 .
H 1 H 6
HO 4 H
.
CH 2OH O.
HO 2 5 ..
OH .. H 6
3 4 1 H
..
H .O. H CH 2OH .O
HO .. 5 .
2 OH 4 H 1
3 .O. H H 6CH OH . .
H HO 2 2 .O.
3 OH .. 5
4 H 1
H .O. H
n
- (1 4) -glycosidic linkage HO 2 OH
3 OH
H
Thus, there is almost no branching and these chains are folded in a left
handed helical structure, as given below in Fig.16.1.
 linkage
H
H
..O.. CH 2OH
...
H
..
2O
H O O.
OH ..
CH
H
HO H H
H H
 linkage  linkage
OH 1
4
The helical structure .. ..
H H
.. H
H O OH O..
gives a compact 4
CH 2OH
shape to amylose OH
.. .. HO
molecules. O H
H ...
H O.
OH
H H H
CH2
 linkage
OH OH
4 ... .. H
H O. O..
left handed helix
Fig. 16.1: Amylose: helical structure, left handed helix
72
SAQ 7
What kind of interactions are responsible for helical structure?
Similarly, the hydrolysis and other reactions along with the molecular weight
determination studies of amylopectin showed that it has a branched structure.
Its molecular weight ranges between 1 to 6 million.
Thus, amylopectin contains D-(+)-glucose units linked by α(1→4) glycosidic

linkages similar to that in amylose but in addition, it has several branches
which have 20-25 glucose units. Such branches are linked by α(1→6)
linkages, i.e. C-1 hydroxyl of the branch forms a glycosidic linkage with the
hydroxyl group at C-6 position of the main chain. This is shown below in
Fig. 16.2 illustrating the part structure of amylopectin.
H 6
CH 2OH . ..
5 O.
O 4 H H
H 1  linkage
HO 2
3 H 6
OH CH 2OH . ..
H . .. 5 O.
O 4. H H
H 1  linkage branch
HO 2 6
3 H CH OH . .
OH 2 .
H . .. 5 O.
O. 4 H
H
HO 1 H
3 2
OH H 6
H CH 2OH . ..
H 6 . . O.
.O . 4 5 Glycogen, the energy
CH 2OH . .. H
.
O 4 5 O H 1 H  linkage
H
H
1
H  linkage HO
3
2 source present in the
HO OH
3
2 H 6
CH 2OH . ..
H cells of animals has
OH ..O ..
H . .. 5 O.
O. 4 H H highly branched
H 1
HO H 6 structure. It has
2 CH 2 . ..
3 OH . .. 5 O. th
H O. 4 H H  linkage branches at every 6
H 1
HO
3 2 H 6
CH 2OH . ..
unit. It also has high
OH
main Chain 5 O.
H . ..
O . 4 H H molecular weight, i.e.
H 1
HO
3
2 about 100 million.
OH . ..
H O.
Fig. 16.2: Amylopectin (part structure).
Thus, amylopectin has highly branched structure having several hundred short
chains of 20-25 D-glucose units which are interconnected. This is shown below
in the Fig. 16.3.
Fig. 16.3: Branched Structure of Amylopectin.
SAQ 8
Write the part structure of amylopectin in Haworth projections.
73
16.3.2 Cellulose
Cellulose forms the main structural material in plants and gives them the
rigidity. For humans, the important uses of cellulose include the structural
material in the form of wood for buildings, cotton and rayon for making clothes,
Cotton is almost pure paper and packaging. Some of the derivatives of cellulose include cellulose
cellulose. trinitrate (gun cotton) which is used in explosives and cellulose xanthate from
which cellulose can be regenerated to give rayon fibre. The treatment of
regenerated cellulose with glycerol softens it and yields the cellophane sheets.
Cellulose is tasteless and insoluble in water. The molecular weight for

cellulose ranges between 2,50,000 to 1,000,000 or even greater than this.
Various chemical reactions and analysis by physical methods indicated that in
cellulose, the D-glucopyranoside units are linked by β-glycosidic linkages at
1→4 positions forming long unbranched chains. The structure thus formed
can be written as follows:
6 CH
2OH
H 5 . ..
 - glycosidic linkage O .OH
4 H 1
OH H
H
6 CH . ..
2OH 3 2
5 . .. O. H OH
H O.
4 H 1
OH H
H
6 CH
2OH
. .. 3 2
. .. O. H OH
5
H O.
4 H
OH 1 n
H
HO H
3 2
H OH
Note that in the β-glycosidic linkage, as shown in the Haworth structure

above, -OH group at C-1 of one D-(+)-glucose unit which is shown upside,
forms the linkage with the -OH group at C-4 of another D-(+)-glucose
molecule which is placed down. This is clearly evident in the part structure of
cellulose shown above. Alternatively, β-glycosidic linkages are also
represented in the way they are shown in the following part structure of
cellulose.
6  linkage
CH2OH
5 .. 6
H CH2OH
O
. . .. 6
4 H
5 CH2OH
OH H 1 . .. H O.. ..
H O. 4 H
5
HO .. H O
. . OH
3 2 OH H 1
H OH H .. 4 H
O
OH H 1
3 2
H OH 2
H
3
n H OH
When represented as the chair conformation, the above structure can be

represented as follows:
H 6
H 6 CH2OH . ..
H 6 CH2OH . .. 5 O. . ..
H 6 CH2OH . .. 5 O. . .. 4 H O.
6 . . 5 O. . .. 4 H O . HO H 1
H CH 2 OH . . H 1
CH2OH . .. 5 O. . .. 4 H O HO 2
. 4 H 1 2 3
5 O . .. H O . HO 3
OH
. .. 4 H O . HO H 1 2 OH H H
O . HO H 1 3 OH H
3 2 H
2 OH H H
3 OH H
H H
H
 - glucosidic linkage
74
Another way of drawing the above part structure is by turning alternate
pyranose rings of above structure by 180o to give the following representation
for cellulose.
Scienceinschool.org/content/cellulose-trees-treats
H6 H H 6
CH 2OH . .. 3 CH 2OH . ..
O. OH
5 2 5 O. ..
. .. 4 H . . HO H 1 .. 4 H O. .
O . HO H 1 O.. 4 5 H. . O. . HO H 1
2 O 2
3 .. 3
OH OH
H H H CH
6 2
OH
H H
Note that by turning the alternate ring, the positions of -OH and -CH2OH
groups do not change and they remain at equatorial position in both the
representations. You can also see that in the above structure, -OH groups on
alternate rings are placed on either side of the chain and hence, are uniformly
distributed on both the sides of the chain. When such long chains lie side by
side, these -OH groups form hydrogen bonds as shown below in Fig. 16.4.
H H H6 H H H6
3 OH CH2OH . .. OH CH2OH . .
. .. 3 .
2 5 O. 2 . .. 4
5 O.
. .HO H 1 O. 4 H HO 1 O
. H ..
. . .. H
O. 4 5 H. .
. HO H 1 O. 4 5 H. .. HO H 1 O..
O. 2 O. 2
3 OH 3
H 6CH2OH H H H 6CH2OH H
OH
H
OH
H H6 H H 6
OH CH2OH . .. CH2 . ..
3 1 .. 3 OH
2 .. 5 O. 2 . .. 5 O.
. ..HO H O 4 H . .. HO H 1 O. 4 H . ..
O. 4 H H 1 H. .
.. HO O. 4 5
. HO H 1 O.
O
5 .. 3
2 O. 2
H CH2OH OH 3 OH
6 H H H CH2OH H H
6
Fig.16.4: Hydrogen bonding between the chains
Many such chains, say about 40, lie laterally to form a sheet. The hydrogen
bonding takes place between the hydroxyl groups across the sheets also.
Such sheets stack and give a rigid, highly insoluble material which forms cell
wall in plants.
6
OH CH2OH H
H .. 3 1 .. H OH
.. 2 . . .. . .. 3 1 ..
O O. 4 O. 2
O.
.
. .. O.
5 . .. 5 . .. 5
O. O. HO 1 O. . ..
4 2 4 O.
CH2OH 3 OH CH2OH
6 H 6
H 6
OH CH2OH H
3 2 1 OH 1 ..
. .. . .. . .. 3 2
HO O. .
. .. O. 4 HO. O.
5 . .. 5 . ..
O. HO 1 O. 5 . ..
O. 2 O.
4 4
3 OH
CH2OH CH2OH
6 H 6
H 6
OH CH2OH . OH H
3 1 4 .. 1
2 . .. 3 . ..
O. 2
HO O. HO O.
. .. 5 H . ..
O. 5 . .. 1 . ..
O. HO O. O.
4 2 5
3 4
CH2OH OH CH2OH
6 H 6
H
6
OH CH2OH H
3 1 4 OH 1 .
2 . .. . .. 3 2 ..
HO O. O. HO .
. .. O
O. 5 . .. 5 . .. 5
HO O. . ..
4 O. O.
2 4
CH2OH 3 OH 1 CH2OH
6 H 6
Fig: 16.5: Sheets showing H-bonding between chains. 75

The side by side arrangement of chains form bundles which twist to give rope
like structures in fibres.
An interesting fact about cellulose is that humans cannot use cellulose as

food whereas animals such as cows and termites can. The reason behind
this is that humans do not have enzymes which can break β(1→4) linkages
whereas symbiotic bacteria in some animals (as mentioned above) provide
such enzymes to them.
16.4 SUMMARY
In this unit, you have learnt the following important concepts.
 Glycosides are formed when the anomeric -OH of a monosaccharide

reacts with alcohols, phenols or other monosaccharide unit.
 Glycosides are named according to the monosaccharide forming them,

i.e. glycoside of glucose is known as a glucoside and that formed by
fructose is called fructoside and so on.
 Disaccharides are formed when two monosaccharide units are linked by a

glycosidic bond.
 Disaccharides may be formed by the combination of the same

monosaccharides or by two different monosaccharides.
 Maltose has two D-(+)-glucose units linked by an α-(1→4) linkage and it is

a reducing sugar;
 In cellobiose, the two D-(+)-glucose units are joined by a β-glycosidic

linkage.
 Lactose, present in the milk of mammals has a D-glactose unit linked to a

D-glucose unit by a β-glycosidic linkage. It is also a reducing sugar.
 Sucrose, the table sugar is a non-reducing sugar.
 In sucrose the D-(+)-glucose unit is linked by α-glucosidic bond to a

fructofuranoside unit which is linked by the β-glycosidic bond.
 Polysaccharides, calledglycans are important both for plants and animals;

their examples being starch, cellulose and glycogen.
 Starch consists of amylose (10-20%) and amylopectin (80-90%).
 Both amylose land amylopectin have D-(+)-glucose units linked to each

other by α-(1→4) glycosidic linkages. The difference between the
amylose and amylopectin is that the amylose does not have any branches
but amylopectin has branching at C-6 -OH group to which the glucose unit
of the branch is joined by the α-glycosidic linkage.
 Amylose can form the helical structure but amylopectin is having a

branched structure and has no helices.
 The pictorial representation of amylose, amylopectin and glycogen can be

shown as given below.
76
 Cellulose forms the structural material in plants which gives them rigidity.
 Cellulose is also a polymer of D-(+)-glucose unit in which these units are

linked by β-(1→4) linkages. The resulting long chains lie side by side to
form bundles which have H-bonding between different chains.Many such
bundles form sheets. These sheets are stacked one over the other to
form the structural materials in plants.
 Thedifferent properties of the disaccharides and polysaccharides arise

due to their different structural features.Thus, their functions can be
correlated to their properties and structures.

1. Write the structure of α-anomer of (+)-maltose and give its name.
2. Draw the structures for the conversion of α-form of maltose to β-form of
maltose.
3. Write the β-anomer of (+)-cellobiose in chair form.
4. Write the chair representation of α-lactose. Also give its name.
5. Why is a double headed arrow placed between 2 and 1positions in the
shorthand name of sucrose?
6. How are amylose and amylopectin different from each other?
7. What is the difference between amylose and cellulose?

8. Name the monosaccharide which forms cellulose. What type of linkages
joins these monosaccharide units?
9. Which carbohydrate is represented by the following part structure?
6 6
CH 2OH CH 2OH
5 . ..
5 . .. H H
H O. H O. H
H
.. 4 1 4
O OH H .. OH H 1  -(1,6)-linkage
.. O .. O..
2
.. 3 2
3
H OH H OH 6
6 6 6
CH2 OH CH 2 CH2 OH
CH2 OH
. ..
5 . .. 5 . .. H5 . .. H H5 O. H
H O. H
H O. H H
O. H
H H 4 ..
.. 4 1 4
H 1 4
OH H
1
.. OH H 1
OH H .. OH .. .O.
.O. O
.. O
.. O
..
2 3 2
3 2 3 2 3
H H OH H OH
H OH OH
 -(1,4)-linkage 77
16.6 ANSWERS
1. Yes
2. Structure of maltonic acid

6 CH 6
2OH
. .. CH2OH
..
H 5 O. H H . .H
O 5
4 H 1 .. H COOH
OH H OH H 1
O
.. 4
HO
3 2 3 2
H OH H OH
3. 4-O-(β-D-glucopyranosyl)-β-D-glucopyranose
4. Howarth projection for α-lactose:

6
CH2OH
5 . ..
6
H O .H
CH2OH 4 H
. .. OH H 1
5 . .. OH
HO O.
O. 3 2
4
H H OH
OH H 1
H H
3 2
H OH
β-D-galactopyranosyl-(1→4)-α-D-glucopyranose
5. β-D-galactopyranosyl-(1→4)-α-D-glucopyranose
or β-D-Galp-(1→4)-α-Glcp.
6. (C6H10O5)n– since all of them contain the hexose, D-(+)-glucose.
7. The hydrogen bonding between the hydroxyl groups stabilises the helical
structure.
8. part structure of amylopectin in Haworth projections:
6 6 6
CH 2OH CH 2OH CH 2OH
5 . .. 5 . .. 5 . ..
H O .H HH O.H H O.H
4 H 4 1 4 H
.. OH H 1
.. OH H .. OH H 1
O
.. O O . ..
.. .. (1,6) glucoside linkage
3 2 3 2 3 2 O.
H OH H OH H HO
glucoside linkage
6 6 6 6 6
CH2OH CH2OH CH2OH CH2 CH2OH
5 . .. 5 . .. 5 . .. 5 . .. 5 . ..
H O .H H O .H H O .H H O .H H O .H
H H H H H
4 1 4 4 4 4
. .. 1 1 1 1
OH H .. OH H .. OH H .. OH H OH H ..
O. ..
O
.. O
.. O
.. O O
..
3 2 3 2 3 2 3 2 .. 3 2
H OH H OH H OH H OH H OH
Structure of amylopectin
78
Terminal Questions
1. H 6CH OH
2 . ..
5 O.
HO
4 H
H H  anomer of glucose,
HO H 6
2 the aglycon
3 OH 1 CH 2OH . ..
O .
H . . 5
4 H
.O. H
HO H 1
 (1,4) - glycosidic bond 3
2
OH
H OH
Name: 4-O-(-D-glucopyranosyl)--D-glucopyranose
 glycosidic
2. linkage glycosidic
linkage
6 CH 2OH 6 CH 2OH
6 CH 2OH 6 CH 2OH
5 . .. . ..
H O. H H 5 O. H 5 . .. H 5 . ..
H 1 4 H H O.1 H O . OH
4 H H 1 H 4 H 1
OH . .. OH 4 H
HO . OH
OH H . .. OH
O HO O. H  orientation
3 2 3 2
H OH H OH 3 2 3 2
H OH H OH
 maltose  orientation
 maltose
6 CH2OH 6 CH2 OH
5 . .. H 5 H
HH O. H OH 1
. ..
4 OH 4 H C
O.
H 1 . .. OH H
HO O.
3 2 3 2
H OH H OH
aldehyde intermediate
3. H 6
 - (1, 4) - glycosidic bond
H
CH 2OH
... 4 6
HO 4 5 O. ... CH 2OH
H O.
H 5
HO
2
1
HO ...  - anomer of glucose,
HH
3 OH
3 O. (the aglycon)
H H H
2
HO 1 OH
 - D - glucopyranosyl unit
H
4. 6
HO CH OH . . HO 6
2 . CH 2OH . ..
5 O. . .. 4 5 O.
4 H H
H O. H
HO H 1 HO H 1
2 2
3 OH 3 OH
H H H OH
- lactose
 -D-galactopyranosyl-(1 4)- -D-glucopyranose
5. Because these positions are linked by the glycosidic bonds formed by the
anomeric hydroxyls placed at these positions of the respective
monosaccharide units linked in that order.
6. Amylose has D-(+)-glucose units linked by α-(1→4) glycosidiclinkages with

no branches. But, in amylopectin, branches occur at every 20-25 units and
are linked by α-(1→6) linkages.
79
7. In amylose, D-(+)-glucose units are linked by α-(1→4) glycosidiclinkages

while in cellulose, these units are linked by β-(1→4) glycosidic linkages.
8. D-(+)-glucose units. These units are linked by β-(1→4) linkages.
9. The structure represents the part structure of amylopectin.
References:
1. Useful website for polysaccharides – starch and cellulose
pslc.ws/macrog/starlose.htm
80

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Unit 13 Amino Acids and Peptides

Then, the structure of peptides will be illustrated. The synthesis of peptides

Expected Learning Outcomes

 list some amino acids and write their structures;

 give the common and IUPAC names of amino acids;

 discuss the zwitterionic nature of amino acids;

 explain the isoelectric point and importance of electrophoresis;

 give various methods of preparation of 2- amino acids;

 discuss the reactions of amino acids;

 briefly explain the structure of peptides;

 describe the methods of synthesis of peptides using N-protection,

 discuss Merrifield solid-phase synthesis and highlight its importance;

 explain the laboratory detection of amino acids.

13.2 Structure and Physical Properties of Amino

13.2.1 Structure of Amino Acids

These are more often referred as α-amino acids.

Table 13.1: Physical Properties of Amino Acids

Amino Acid, Name Abbreviation Abbreviation

NH2 Three letter One letter

CH3 NH2 valine Val V

CH3 NH2 Ile I

13.2.2 Zwitterionic Nature

13.2.3 Isoelectric Point and Electrophoresis

Table 13.2: pKa and pHi values of some amino acids

You can verify from Table 13.2 that

It is a technique which involves the migration of charged particles using

13.2.4. Stereochemistry of Amino Acids:

According to the older D, L system of specifying the configuration (discussed

COO Enzymes use up one

On the basis of your knowledge about assigning the absolute configuration,

13.3 SYNTHESIS OF 2-AMINO ACIDS

Table 13.3: Methods of Synthesis of 2-Amino Acids

1. By N-alkylation of 1,2-benzenedicarboxylic imide (phthalimide) anion

2-substituted amino acids can also be prepared.

2. From aldehydes : Strecker Synthesis

3. From 2-halo acids

R CHCOOH + NH3 R CHCOOH

13.3.1 Gabriel Phthalimide Synthesis

It involves the N-alkylation of 1,2-benzenedicarboxylic imide (phthalimide)

A variation of the above mentioned method utilises diethyl

13.3.2 Strecker Synthesis

13.3.3 From 2-Halo Acids

2-bromopropanoic acid 2-aminopropanoic acid

13.4 REACTIONS OF AMINO ACIDS

Neutralisation of the hydrochloride with alkali yields the ester.

Esters of amino acids undergo intermolecular cyclisation to yield cyclic amides

2. Alkanoylation of Amino Acids

glycine ethanoic N-ethanoylglycine

3. Reaction with Ninhydrin

When the aqueous solution of a 2-amino acid is treated with triketohydrindene

O& O Ninhydrin test is given

The blue-violet coloured compound formed is also known as Ruhemann’s

This is an important reaction used in the detection of small amounts of amino

13.5 STRUCTURE OF PEPTIDES

General Peptide Structure

Peptides can be classified as dipeptides, tripeptides and tetrapeptides,

A-A, B-B, A-B and B-A

Therefore, for the selective synthesis of a particular peptide, we have to

i) the N-terminal of one amino acid and