BIOLOGY FOR LIFE SCIENCES-I

2022-2023, 1ST SEMESTER

MAJOR ORGANIC CONSTITUENTS OF THE CELL

1 INTRODUCTION

A typical cell is an incredibly complex array of chemical molecules. Moreover, cells manufacture
variety of organic compounds. The main types of compounds contained in and synthesized by
cells are carbohydrates, lipids, proteins and nucleic acids. Some of these are used as components
of cell parts; other provide energy for cellular activities; and still others are of importance for
the regulation of cellular chemical activities (metabolism). The main chemical components of
cells are structures called macromolecules. Macromolecules are built up of individual ‘building
blocks’ which are connected in specific ways. A single building block is called a monomer and
the macromolecule is called a polymer and the process of connecting the monomers together is
called polymerization. There are only a few types of polymers important in cell biochemistry,
and each is made up of a characteristic of monomers. Monomers are organic compounds
containing up to 30 carbon atoms and are grouped in classes according to their chemical
properties. There are four classes of monomers to be considered here: sugars, the monomeric
constituents of polysaccharides; fatty acids, the monomeric units of lipids (or fats); amino acids,
the monomeric constituents of the proteins; and nucleotides, the basic units of nucleic acids
(DNA or RNA).

1.1 CARBOHYDRATES
Carbohydrates (sugars), also known as saccharides are organic compounds containing carbon,
hydrogen and oxygen usually in the ratio 1:2:1. The structural formula for glucose, the most
abundant sugar is C6H12O6 (Fig 1). The most biologically relevant carbohydrates are those
containing 4, 5, 6 and 7 carbon atoms. The C5 sugar (pentose), ribose and deoxyribose, are of
special significance because of their roles as the structural backbones of the nucleic acids. C6
sugars (hexoses) are the monomeric constituents of cell polymers (glycogen, cellulose, starch)
and energy reserves.
 Figure 1. The structural formula for glucose and fructose

Figure 2. Formation of α and β- glycosidic bonds.

Polysaccharides are defined as high molecular weight carbohydrates containing many

(hundreds or thousands) monomeric units connected to one another by a type of covalent bond
referred to as a glycosidic bond through a condensation reaction (Fig 2). The glycosidic bond can
exist in two different orientations, referred to as alpha (α) and beta (β) as indicated in Figure 2.
If two sugar units (monosaccharides) are joined by a glycosidic linkage the resulting molecule is
called a disaccharide. The addition of one more monosaccharide would yield a trisaccharide,
several more an oligosaccharide and an extremely long chain of monosaccharides in glycosidic
linkages is called a polysaccharide (Fig 3). Polysaccharides can be broken down into their
constituting monosaccharides by hydrolysis (through addition of water molecule and breakage
of the glycosidic bonds), which can be achieved enzymatically or chemically.

Figure 3. General structure of some common polysaccharides

1.2 LIPIDS
Like carbohydrates, lipids a second major group of biological compounds are composed
principally of carbon, oxygen, and hydrogen but they may also contain other elements
particularly phosphorus and nitrogen. Unsubstituted lipids, being non-polar, are relatively
insoluble in water, but soluble in organic solvents such as ether. Fats among the best-known
lipids are the natural fats. Important as energy-storage molecules in living organisms, the fats
also provide insulation, cushioning and protection for various part of the body. Each molecule of
fat is composed of building-block compounds: an alcohol called glycerol and fatty acids. Glycerol
(also sometimes called glycerin) has a backbone of three carbon atoms, each carrying a hydroxyl
(-OH) group (Fig 4). Fatty acids, like all organic acids, contain a –COOH group (called a carboxyl
group). Organic acids and alcohols have a tendency to combine via ester bond through
condensation reactions (elimination of water molecule). Since glycerol has three alcoholic
groups, it can combine with three molecules of fatty acid to form a molecule of fat (Fig 4). Hence
fats are sometimes also called triglycerides.
 Figure 4. Synthesis of a fat.

1.3 PROTEINS
Far more complex than either carbohydrates or lipids, proteins are fundamental to both the
structure and function of living materials. Like carbohydrates and lipids, proteins are composed
of relatively few, simple building block compounds. Amino acids are the monomeric units of
proteins. Most amino acids consist only of carbon, hydrogen oxygen and nitrogen, but two of the
20 common amino acids found in the cell also contain sulfur atoms as well. All amino acids (Figure
5) contain two important functional groups, a carboxyl group (-COOH) and an amino group (-
NH2). The groups are functionally important because covalent bonds between the carbon of the
carboxyl group and nitrogen of the amino group (with elimination of a water molecule i.e.
condensation) forms the peptide bond, a type of covalent bond characteristic of proteins (Fig 6).

Figure 5. General structure of an amino acid ‘R’ indicates the side chain.

Proteins are comprised of linear polymers of various lengths containing defined sequences of
amino acids covalently bonded by peptide linkages. Two amino acids connected together
constitute a dipeptide, three amino acids a tripeptide and so on. Many amino acids covalently
linked via peptide bonds constitute a polypeptide. Proteins consist of one or more polypeptides.
The number of amino acids varies from one protein to another. Proteins with as few as 20 and
 as many as 10.000 amino acids are known. Since proteins may vary in their composition,
sequence, and number of amino acids, it is easy to see that enormous variations in protein types
is possible. Proteins play key role in cell function. Two kinds of proteins are recognized in cells:
Catalytic proteins (enzymes) and Structural proteins. The proteins called enzymes serve as
catalysts for the wide variety of chemical reactions which occur in the cell. Structural proteins
are those which become integral parts of the structures of cells, in membranes, cell walls and
cytoplasmic components. In essence, a cell is what it is because of the kinds of proteins which it
contains.

Figure 6. Peptide bond formation.

1.4 NUCLEIC ACIDS

The nucleic acids, DNA and RNA, are polymers of monomers called nucleotides. DNA and RNA
are thus both polynucleotides. DNA carries the genetic blueprint for cell and RNA acts as an
intermediary molecule to convert the blueprint into defined protein sequences. Despite their
important functions, nucleic acids are composed of a relatively few simple building blocks. Each
nucleotide is composed of three separate units: a five-carbon sugar, a nitrogen base and a
molecule of phosphate as shown in Figure 7.
 Figure 7. General structure of a nucleotide

Nucleic acids are long polymers in which nucleotides are covalently bonded to one another in
a defined sequence. The backbone of the nucleic acid is a polymer in which sugar and
phosphate alternate (Fig 8). In biological terms nucleic acids are composed of nucleotides
covalently attached via phosphate residues from carbon 3 of one sugar to carbon 5 of the
adjacent sugar. The phosphate linkage is chemically a phosphodiester, since a single
phosphate is connected by ester linkage to two separate sugars.

Figure 8. Structure of a part of a DNA chain.

2 OBJECTIVES

After completing this exercise, you will be able to:

1. Define reducing and non-reducing sugars, condensation and hydrolysis
reactions, monomer, polymer, glycosidic bond, ester bond, phospodiester bond,
peptide bond, polysaccharide, polypeptide, glycerol, fatty acid, amino acid,
 nucleotide, polynucleotide
2. Discuss the chemical properties and the main functions of carbohydrates,
proteins, lipids and nucleic acids.
3. Identify sugars, proteins and lipids by simple test
4. Demonstrate the presence of sugars, proteins and lipids in biological materials.

3 MATERIALS AND METHODS

A detailed chemical analysis of living matter in a task of considerable technical and theoretical
difficulty, especially when it is desired to determine not only what specific compounds are
present but also precisely how much of each. However, if the analyst is not after detail and exact
quantity but wishes merely to identify the general classes of cellular compounds; he can
successfully employ a number of rather simple tests. Several such tests will be examined and
used in the following exercises.

3.1 TEST FOR SIMPLE SUGAR

Benedict’s Test: This test is based on the reducing property (the presence of free aldehyde or
keto group) of sugars. This test is performed under mildly alkaline conditions and is a very
sensitive test. Monosaccharides and some disaccharides which possess a potential aldehyde or
keto group are called reducing sugars. Such sugars will reduce the cupric ions of copper sulfate
solution (Benedict’ Reagent) and an insoluble cuprous oxide formation will serve as an indication
for a positive test. The color and density of the precipitate (cuprous oxide) gives a rough
estimation of the amount of sugar present. A green precipitate means relatively little reducing
sugar, a brown or red precipitate means more reducing sugar is present.
Test-1:
1. Place in a test tube 2 ml of a 1 % (w/v) glucose solution and 5 ml of Benedict’s reagent.
Shake and heat the contents of the tube in a boiling water bath on hot plate for 2 minutes,
and allow the tube cool.
2. Observe the color and texture of the precipitate formed.
Test-2:
1. Place in a test tube 2 ml of a 1 % (w/v)
(1) distilled water (control)
(2) sucrose
(3) starch
 solution and 5 ml of Benedict’s reagent. Shake and heat the contents of the tube in a
boiling water bath on hot plate for 2 minutes, and allow the tube cool.
2. Observe the color and texture of the precipitate formed.
Test-3:
1. Cut a cube of apple (1 cm3), grind it up in a mortar with a small quantity of water.
2. Transfer the material to a test tube and add water to make a total volume of about 5 ml.
3. Add an equal volume of Benedict’s reagent and heat in boiling water bath on hot plate and
allow the tube cool.
4. Observe the color and texture of the precipitate formed.
Test-4:
1. Place 5 ml of starch suspension in a test tube and add 1 ml of 10 % (w/v) HCl.
2. Place the tube in a boiling water bath on hot plate for 10 min. (Remember that starch is a
polymer of glucose units interconnected by glycosidic bonds and this chemical treatment
will help in breaking these bonds and releasing free glucose)
3. Let the tube to cool and neutralize the solution by the addition of 1 ml of 10 % (w/v) NaOH
solution. Test the solution with Benedict’s reagent as before.
4. Compare and discuss the results obtained in these two tests.

3.2 TEST FOR LIPIDS

To test for the presence of fats or lipids, place the test material in water in a test tube, and add a
drop of Sudan III stain. The Sudan dyes are highly soluble in alcohol, as well as in fats and oils of
all types, but much less in water. When an alcoholic solution of the dye is added to the water
preparation of material to be examined, the dyes quickly concentrated in any oil droplets
present.
3.3 TEST FOR PROTEINS
The Biuret Test: This test is very sensitive and important for proteins and for polypeptides
generally. The biuret test should always be accompanied by a control test in which distilled
water instead of protein is used, every other step being identical. Note that a positive result for
proteins is a distinctly violet color; a blue color is NOT indicative of proteins. The color of biuret
appears to be due specially to the atomic grouping of the peptide bond, hence the biuret test will
be positive for compounds containing such groupings. This is the case for proteins, which
contain peptide linkages. Actually, the biuret test is sensitive only for polypeptides longer than
dipeptides.

Figure 9. Biuret-urea formation

1. To 2 ml egg albumin (egg white) add 3 ml of 10 % (w/v) NaOH solution and shake well.
2. Then add several drops of 0.5 % (w/v) CuSO4 solution and mix. A purple color will
develop.
3. This test is named after the compound BIURET, formed by the union of two molecules of
urea.