Introduction to

Bioscience and
Technology
(ID2L002)
L-T-P-C: 2-0-0-2

Dr. Soumendra Rana

IIT Bhubaneswar Image Credits: Books, Journals, Google
4 THE NATURE OF BIOTECHNOLOGY

Fig. 1.1 The interdisciplinary

nature of biotechnology.

Bioscience and
Biotechnology ?

The integrated use of both

basic and engineering
sciences in order to achieve
technological or industrial
application capabilities, by
recruiting various agents
of biological origins are
generally regarded as
“Biotechnology”.

Unlike a single scientific discipline, biotechnolog

wide array of relevant fields, such as microbiology, bioc
biology, cell biology, immunology, protein engineerin
 Biotechnology Industries
THE NATURE OF BIOTECHNOLOGY

Table 1.3 Types of companies involved with biotechnology

Therapeutics Pharmaceutical products for the cure or control of human diseases,

including antibiotics, vaccines, gene therapy.
Diagnostics Clinical testing and diagnosis, food, environment, agriculture.
Agriculture/Forestry/Horticulture Novel crops or animal varieties, pesticides.
Food Wide range of food products, fertilisers, beverages, ingredients.
Environment Waste treatment, bioremediation, energy production.
Chemical intermediates Reagents including enzymes, DNA/RNA, speciality chemicals.
Equipment Hardware, bioreactors, software and consumables supporting
biotechnology.

Biotechnology is a demanding industry that requires a skilled work-

force and a supportive public to ensure continued growth. Economies that
encourage public understanding and provide a competent labour force
should achieve long-term benefits from biotechnology. The main types of
How to
Understand a
Biological
System
Better?
How Different Are They?
Molecular
Complexity
of Organisms
 Cell: The Building Block

• Cells Are the Structural and Functional Units of All Living Organisms.

• All cells have, for at least some part of their life, either a “nucleus” or a
“nucleoid”, in which the genome — the complete set of genes, composed of
DNA—is stored and replicated.

• Cells with nuclear envelopes are called eukaryotes.

• Cells without nuclear envelopes (bacterial cells) are called prokaryotes.

• Animal and plant cells are typically 5 to 100 μm in diameter, and many
bacteria are only 1 to 2 μm long.
 E. Coli vs. Others

• The DNA that is the entire genetic material of E. coli is a single

molecule containing 4.64 million nucleotide pairs.

• An embryo with all its 35,000 genes, is constructed of 3 billion

nucleotide pairs, intact.
Cell: The
Building
Block
Molecular
Components
of E. Coli
Cell

Escherichia coli
The Biomolecules

Lipids

Sterols
The Five Senses of
Life
• Biopolymers like Proteins control myriad of function in
Human body
Proteins ?

• Functionally versatile biological

macromolecules made of one or more chains
of amino acids. A short chain of amino acids is
often called as polypeptide
• Protein has two components: main chain &
side chain
• In proteins amino acids are joined by an amide
/ peptide bond
Main chain • Amino acids are usually L-chiral in proteins,
whereas Sugars are D-chiral in DNA
• Stereochemistry defines and critically limits
Protein the scope of conformational possibility
(limited morphology) in proteins of poly-L
amino acids
• “Rod” and “Flat ribbon” like morphology;
Side chain ~1000 topological variants.
Amino acids ?
• Organic molecule having, Amine
(-NH2) and Carboxylic (-COOH)
functional group. Otherwisely
called as a-amino carboxylic acid
• Name and property of amino
acids change with different R-
groups
• 23 amino acids are found in
proteins; 20 are standard amino
acids (genetic code) and 3 are
non-standard amino acids (post
translational modification)
• Out of 20 standard amino acids,
19 are L-chiral, 1 is achiral. D-
chiral amino acids are found in
some bacterial species Cahn-Ingold-Prelog rules
 Nutrition Synthesized in the Body

Classification
of Standard
Amino acids

Non-standard
1. Pyrrolysine
2. Selenocysteine
3. N-Formylmethionine
 • Simplest Peptide
Glycylglycine • Emil Fischer and Ernest Fourneau in 1901
The Building Blocks of Proteins
Types of
Amino acids
Small Amino acids

Non-polar aliphatic R-groups

Hydrophobic Amino acids

Non-polar aliphatic R-groups

Nucleophilic Amino acids

Polar uncharged R-groups

Amino acids with Amides

Polar uncharged R-groups

Basic Amino acids

Positively charged R-groups

Acidic Amino acids

Negatively charged R-groups

Aromatic Amino acids

Aromatic R-groups
Classification based on Metabolic Fate

q Glucogenic amino acids: Amino acids

that serve as precursors for biosynthesis
of glucose (Gluconeogenesis).

q Ketogenic amino acids: Amino acids

that breakdown to form ketone bodies.

q Both glucogenic and ketogenic amino

acids: Amino acids that breakdown to
form precursors for both ketone bodies
and glucose.
Isoelectric Point (pI)

Anionic Form Cationic Form

 Isoelectric Point (pI)

• The isoelectronic point (pI) is the pH at which the amino acid

does not migrate under the influence of an electric field.
• pI is the pH at which the amino acid is neutral, i.e. the zwitterion
form is dominant.
• The pI is given by the average of the pKas that involve the
zwitterion.
 Example

Amino acids with neutral side chains: Characterised

by two pKas : pKa1 (-COOH) and pKa2 (-NH2). The pI
will be the average of these two pKas, i.e. pI = 1/2
(pKa1 + pKa2).

q At very acidic pH (below pKa1) the amino acid will

have an overall +ve charge and at very basic pH
(above pKa2 ) the amino acid will have an overall -ve
charge.

q For amino acid like Glycine (G), pKa1= 2.34 and

pKa2 = 9.6. Thus, pI = 5.97
 Example

Amino acids with acidic side chains: Introduction of a third acid dissociation constant : pKa3 (-R
group)

q The pI will be at a lower pH because the acidic side chain introduces an "extra" negative charge. So the
neutral form exists under more acidic conditions when the extra -ve has been neutralised.

q For amino acid like aspartic acid (D), the neutral form is dominant between pH 1.88 and 3.65. Thus, pI
= 1/2 (pKa1 + pKa3) = 2.77.
Example

Amino acids with basic side chains : pKa3 (-R group)

q Histidine (H) has three acidic groups of pKa's 1.82 (-COOH), 6.04 (pyrrole -NH)
and 9.17(ammonium-NH).
q Depending on the pH [from acidic pH (top) to basic pH. (bottom)], Histidine can
exist in four forms.
q As base is added, the most acidic proton is removed first (-COOH), then the pyrrole
NH then finally the amino NH.
q At pH < 1.82, A is the dominant; B is dominant between 1.82 < pH < 6.02; C is
dominant between 6.02 < pH < 9.17, and when pH > 9.17, D is the major form in
solution.
q The pI of Histidine will be at a higher pH, as the basic side chain introduces an
"extra" +ve charge. So the neutral form exists under more basic conditions when the
extra +ve has been neutralised.
q Thus, for Histidine, the pI = 1/2 (pKa2 + pKa3) = 7.59
 Methyl Red (MR)

HMR= [H+] + [MR-]

The acid dissociation constant K can be given by
Ka = [H+] [MR-] / [HMR]

The negative log of the above equation will provide

pKa = pH – log [MR-] / [HMR]

• HMR form has a bright red colour in acidic solution, which changes to yellow in its basic form
MR-. Both HMR and MR- absorb strongly in the visible range. Thus, the ratio of [MR-]/[HMR]
can be determined spectroscopically, which will help in calculating the pKa of MR at a
given pH.
Properties of
Amino acids
Electrophoresis
Why study proteins at pH ~7.4?

q At a pH higher than the pKR, the carboxylic side chains

of the amino acids lose the H+ ion and thus can
maintain a net negative charge.

q Similarly, at a pH lower than the pKR, the aspartic acid

and glutamic acid side chains can remain uncharged.

q At a pH lower than the pKR, the lysine, arginine and

histidine side chains accept an H+ ion and thus can
maintain a net positive charge.

q At a pH higher to their respective pKR, the amine side

chains are uncharged.

q Helps in achieving the overall neutrality of the system.