Biotechnology

Humans have been making use

of biotechnology since they
discovered farming. This use
N
oo

extended from the planting of

n
Ac
ad

seeds to the control of plant

em
y

growth and crop production.

These techniques were used to
enhance product quality and
to meet specific requirements.

Page 1 of 70
 INTRODUCTION OF
BIOTECHNOLOGY
Biotechnology is defined as
the use of living organisms
N

in processes for the

oo
n
Ac

manufacture of useful
ad
em

products or for services.

y

Although the term

biotechnology is new, the
discipline itself is very old.

Page 2 of 70
 Example of natural or old
biotechnology
Fermentation and other
such processes, which are
N
oo
n

based on the natural

Ac
ad

capabilities of organisms,
em
y

are commonly considered

as old biotechnology.

Page 3 of 70
 Genetic engineering
Genetic engineering i.e. the
artificial synthesis,
modification, removal,
addition and repair of the
N
oo
n

genetic material (DNA) is

Ac
ad

considered as modern
em
y

biotechnology. It is done to
alter the characteristics of
organisms.

Page 4 of 70
 History of genetic
engineering
The work on genetic
engineering started in
1944 when it was proved
that DNA carries the genetic
N
oo

information. Scientists
n
Ac
ad

isolated the enzymes of DNA

em
y

synthesis and then prepared

DNA outside cells. In 1970s,
they were able to cut and
paste the DNA of organisms.

Page 5 of 70
 Human Insulin Project
In 1978, scientists
prepared human insulin by
inserting the insulin gene in
N
oo
n

bacteria. Human growth

Ac
ad

hormone was also

em
y

synthesized in bacteria.

Page 6 of 70
 Human Genome Project
In 1990, the Human
Genome Project was
launched to map all the
N
oo
n

genes in human cell. The

Ac
ad

complete map of human

em
y

genome was published in

2002.

Page 7 of 70
 Important information
In Scotland, in 1997, an
embryologist Ian
N
oo
n

Wilmut produced a sheep

Ac
ad

(Dolly) from the body cell of

em
y

an adult sheep.

Page 8 of 70
 Scope and Importance
of Biotechnology
In recent years,
biotechnology is growing as a
separate science. It has
attracted the attention of
N
oo

many intellectuals from

n
Ac
ad

diverse fields like agriculture,

em
y

medicine, microbiology and

organic chemistry. The scope
for biotechnology is so wide
that it is difficult to
recognize the limits.
Page 9 of 70
 Biotechnology in the
Field of Medicine
In the field of medicine,
biotechnologists synthesized
insulin and interferon
(antiviral proteins) from
N
oo
n

bacteria and released for

Ac
ad

sale. A large number of

em
y

vaccines and antibodies;

human growth hormone
and other medicines have
also been produced.

Page 10 of 70
 Biotechnology in the
Field of Medicine
Various enzymes are being
synthesized for medicinal as
well as industrial use. Gene
therapy (treatment through
N
oo
n

genes) has become

Ac
ad

important in recent years.

em
y

Biotechnology also proved

much beneficial in forensic
medicine. The study of DNA
helps in the identification of
criminals.
Page 11 of 70
 Biotechnology in the
Field of Food
Fermented foods (e.g.
pickles, yogurt), malted
foods (e.g. powdered milk: a
mixture of barley, wheat
N
oo
n

flour and whole milk),

Ac
ad

various vitamins and dairy

em
y

products are produced by

using microorganisms. Wine
and beer are produced in
beverage industry.

Page 12 of 70
 Biotechnology in the
Field of Agriculture
Biotechnology has also
revolutionized research
activities in the area of
agriculture. Transgenic
N
oo
n

(organisms with modified

Ac
ad

genetic set-up) plants are

em
y

being developed; in which

desirable characteristics are
present e.g. more yields and
resistance against diseases,
insects and herbicides.
Page 13 of 70
 Transgenic animals
Transgenic goats, chickens,
cows give more food and
milk etc. Many animals like
N
oo
n

mice, goats, cows etc. have

Ac
ad

been made transgenic to

em
y

get medicines through their

milk, blood or urine.

Page 14 of 70
 Biotechnology and
Environment
Biotechnology is also being
used for dealing with
environmental issues, like
pollution control,
N
oo
n

development of renewable
Ac
ad

sources for energy, and

em
y

biodiversity conservation.
Bacterial enzymes are used
to treat sewage water to
purify.

Page 15 of 70
 Microbes
Microbes are being
developed to be used as
biopesticides, biofertilizers,
biosensors etc. Such
N
oo
n

transgenic microorganisms
Ac
ad

are also used for the

em
y

recovery of metals, cleaning

of spilled oils and for many
other purposes.

Page 16 of 70
 Fermentation
Fermentation is the process
in which there is incomplete
oxidation-reduction of
glucose. Fermentation has
N
oo
n

been in the knowledge of

Ac
ad

man since centuries, but it

em
y

was believed that it is

purely a chemical process.

Page 17 of 70
 History of fermentation
In 1857, Pasteur convinced
the scientific community
that all fermentations are
the results of microbial
N
oo
n

activity. He showed that

Ac
ad

fermentation is always
em
y

accompanied by the
development of
microorganisms.

Page 18 of 70
 Classification of
fermentation
There are many kinds of
fermentation and each kind
is a characteristic of
N
oo
n

particular microbial group.

Ac
ad

Fermentations are classified

em
y

in terms of the products

formed.

Page 19 of 70
 Carbohydrate fermentation
The initial steps of
carbohydrate fermentation
are identical to those of
respiration. The process
N
oo
n

begins with glycolysis, in

Ac
ad

which the glucose, molecule

em
y

is broken into two

molecules of pyruvic acid.

Page 20 of 70
 Carbohydrate fermentation
and its products
N
oo
n
Ac
ad
em
y

Page 21 of 70
 1. Alcoholic Fermentation
(by yeast)
This fermentation is carried
out by many types of yeast
N
oo
n

such as Saccharomyces
Ac
ad

cerevisiae. This process is

em
y

quite important and is used

to produce bread, beer,
wine and distilled spirits.

Page 22 of 70
 Process of Alcoholic
fermentation
In this process, carbon
dioxide is removed from
pyruvic acid. The product
N
oo
n

i.e. acetaldehyde is then

Ac
ad

reduced to ethanol. The

em
y

carbon dioxide produced

during this fermentation
causes the rise of the bread.

Page 23 of 70
 Alcoholic fermentation
N
oo
n
Ac
ad
em
y

Page 24 of 70
 2. Lactic Acid
Fermentation (by bacteria)
In this process, pyruvic acid
is reduced to lactic acid.
It is carried out by many
bacteria e.g. Streptococcus
N
oo
n

and many Lactobacillus

Ac
ad

species. It is quite important

em
y

in dairy industry where it is

used for souring milk and
also for production of
various types of cheese.

Page 25 of 70
 Lactic Acid Fermentation
N
oo
n
Ac
ad
em
y

Page 26 of 70
 Fermentation In
Biotechnology
In beginning, the meaning of
fermentation process was
the use of microorganisms
for the production of foods,
N
oo
n

beverages (beers, wines) and

Ac
ad

spirits. However, in
em
y

biotechnology the term

“fermentation” means the
production of any product
by the mass culture of
microorganisms.
Page 27 of 70
 Applications of
Fermentation
In fermentation, maximum
growth of an organism is
obtained for the production
N
oo
n

of desired products of
Ac
ad

commercial value.
em
y

Traditionally, only food and

beverage products were
produced by using
fermentation.

Page 28 of 70
 a- Fermented Foods
Fermentation often makes
the food more nutritious,
more digestible and tastier.
N
oo
n

It also tends to preserve the

Ac
ad

food, lowering the need for

em
y

refrigeration.

Page 29 of 70
 Groups of fermented
foods
Cereal products: Bread is
the commonest type of
N

fermented cereal product.

oo
n
Ac

Wheat dough is fermented

ad
em

by S. cerevisiae along with

y

some lactic acid bacteria.

Page 30 of 70
 Dairy products: Cheese and
yogurt are important
fermentation products.
Cheese is formed when a
milk protein is coagulated.
N
oo
n

This happens when the acid

Ac
ad

produced by lactic acid

em
y

bacteria reacts with milk

protein. Yogurt is made
from milk by different
lactic acid bacteria.

Page 31 of 70
 Fruit and vegetable
products: Fermentation is
usually used, along with salt
N
oo
n

and acid, to preserve pickle,

Ac
ad

fruits and vegetables.

em
y

Page 32 of 70
 Beverage Products: Beer
is produced from cereal
grains which have been
malted, dried and ground
into fine powder.
Fermentation of the powder
N
oo

is done by yeast. This

n
Ac
ad

process breaks the glucose

em
y

present in powder into

pyruvic acid and then into
ethanol. Grapes can be
directly fermented by yeasts
to wine.
Page 33 of 70
 b- Industrial Products
Aspergillus: Formic acid is
used in textile dyeing,
leather treatment,
electroplating, rubber
N
oo
n

manufacture
Ac
ad

Saccharomyces: Ethanol is
em
y

used as solvent; used in the

production of vinegar and
beverages

Page 34 of 70
Saccharomyces: Glycerol is
used as solvent; used in the
production of plastics,
cosmetics and soaps; used
N
oo
n

in printing; used as
Ac
ad

sweetener
em
y

Bacillus: Acrylic acid is used

in the production of plastics

Page 35 of 70
 Fermenter
Fermenter is a device that
provides optimum
environment to
microorganisms to grow
N
oo
n

into a biomass, so that they

Ac
ad

can interact with a

em
y

substrate, forming the

product. Fermentation is
carried out in fermenters,
in the following two ways.

Page 36 of 70
 Batch Fermentation
In this process, the tank of
fermenter is filled with the
raw materials to be
fermented. The
N
oo
n

temperature and pH for

Ac
ad

microbial fermentation is
em
y

properly adjusted, and

nutritive supplements are
added.

Page 37 of 70
 A batch fermenter

Pure
Raw
culture of
material
bacteria
N
oo
n
Ac
ad
em
y

Fermentation

products and

bacterial culture

Page 38 of 70
 Process of batch
fermentation
All the material is steam
sterilized. The pure culture of
microorganisms is added to
fermenter from a separate
N
oo

vessel. Fermentation proceeds

n
Ac

and after the proper time the

ad
em

contents of fermenter are

y

taken out. Fermenter is cleaned

and the process is repeated.
Thus, fermentation is a
discontinuous process divided
into batches.
Page 39 of 70
 Continuous Fermentation
In this process, the
substrate is added to
fermenter continuously at a
fixed rate. This maintains
N
oo
n

the microorganisms in
Ac
ad

growth phase.
em
y

Fermentation products are

taken out continuously.

Page 40 of 70
 A continuous fermenter

Raw

material
N
oo
n
Ac
ad
em
y

Bacterial

culture

Fermentation

products

Page 41 of 70
 Advantages of using
Fermenters
 Controlled environment
 Production of materials
in bulk quantities
N
oo
n

 Optimizes the growth of

Ac
ad

the organisms
em
y

 Inexpensive production
of materials

Page 42 of 70
 Genetic Engineering
Genetic engineering or
recombinant DNA
technology involves the
N
oo
n

artificial synthesis,
Ac
ad

modification, removal,
em
y

addition and repair of the

genetic material (DNA).

Page 43 of 70
 Revolution in genetic
engineering
Genetic engineering
developed in the mid-
1970s when it became
possible to cut DNA and to
N
oo
n

transfer particular pieces

Ac
ad

of DNA from one type of

em
y

organism into another. As

a result, the characteristics
of the host organism could
be changed.

Page 44 of 70
 Becterium as an example
If host organism is a
microorganism, such as a
bacterium, the transferred
DNA is multiplied many
N
oo
n

times as the
Ac
ad

microorganism multiplies.
em
y

Consequently, it is possible
to obtain millions of copies
of a specific DNA inside a
bacterial cell.

Page 45 of 70
 Objectives of Genetic
Engineering
 Isolation of a particular
gene or part of a gene for
various purposes such as
gene therapy
N
oo

 Production of particular
n
Ac
ad

RNA and protein molecules

em
y

 Improvement in the
production of enzymes,
drugs and commercially
important organic
chemicals
Page 46 of 70
 Objectives of Genetic
Engineering
 Production of varieties
of plants having particular
desirable characteristics
N
oo
n

 Treatment of genetic
Ac
ad

defects in higher
em
y

organisms

Page 47 of 70
 Basic Steps in Genetic
Engineering
1. Isolation of the gene of
interest
In the first step, the genetic
engineer identifies the gene
N
oo
n

of interest in a donor
Ac
ad

organism. Special enzymes,

em
y

called restriction
endonucleases, are used to
cut the identified gene from
the total DNA of donor.

Page 48 of 70
2. Insertion of the gene into
a vector
A vector is selected for the
transfer of the isolated gene
of interest to the host cell.
N
oo
n

The vector may be a

Ac
ad

plasmid (the extra-

em
y

chromosomal DNA present

in many bacteria) or a
bacteriophage.

Page 49 of 70
 Recombinant DNA
The gene of interest is
attached with the vector
DNA by using endonuclease
(breaking enzymes) and
N
oo
n

ligase (joining enzymes). The

Ac
ad

vector DNA and the

em
y

attached gene of interest

are collectively called
recombinant DNA.

Page 50 of 70
 3. Transfer of recombinant
DNA into host organism
Recombinant DNA is
transferred to the target
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oo
n

host. In this way, host

Ac
ad

organism is transformed
em
y

into a genetically modified

organism (GMO).

Page 51 of 70
 4. Growth of the GMO
The GMO are provided
suitable culture medium for
N
oo
n

growth to give as much

Ac
ad

copies of the gene of

em
y

interest as needed.

Page 52 of 70
 5. Expression of the gene
The GMO contains the gene
of interest and
N
oo
n

manufactures the desired

Ac
ad

product, which is isolated

em
y

from culture medium.

Page 53 of 70
 Achievements of Genetic
Engineering
Human insulin
Human insulin gene was
transferred into bacteria.
N
oo
n

The genetically modified

Ac
ad

bacteria became able to

em
y

synthesize insulin. Diabetics

are now receiving this
insulin.

Page 54 of 70
 The Human growth
hormone
In 1977 an E. coli
bacterium was created that
N
oo
n

was capable of synthesizing

Ac
ad

the human growth

em
y

hormone.

Page 55 of 70
 The hormone thymosin
The hormone thymosin
which may prove effective
against brain and lung
N
oo
n

cancer has been produced

Ac
ad

by genetically modified
em
y

microorganisms.

Page 56 of 70
 Beta-endorphin
Beta-endorphin, a pain
killer produced by the
brain, has also been
N
oo
n

produced by genetic
Ac
ad

engineering techniques.
em
y

Page 57 of 70
 Vaccines
Genetic engineers produced
a safe vaccine against the
foot and mouth disease (a
viral disease in cattle, goats
N
oo
n

and deer). Similarly many

Ac
ad

vaccines have been

em
y

produced against human

diseases such as hepatitis B.

Page 58 of 70
 Interferons
Interferons are anti-viral
proteins produced by cells
infected with viruses. In
N
oo
n

1980, interferon was

Ac
ad

produced in the genetically

em
y

modified microorganisms,
for the first time.

Page 59 of 70
 The enzyme urokinase
The enzyme urokinase,
which is used to dissolve
N
oo
n

blood clots, has been

Ac
ad

produced by genetically
em
y

modified microorganisms.

Page 60 of 70
 Haemophilia
Now it has become possible
to modify the genes in the
human egg cell. This can
N
oo
n

lead to the elimination of

Ac
ad

inherited diseases like

em
y

haemophilia.

Page 61 of 70
 Blood diseases
Genetic engineering
techniques can also be used
to cure blood diseases like
thalassemia and sickle-cell
N
oo
n

anaemia, which result from

Ac
ad

defects in single genes.

em
y

Normal genes could be

transferred into the bone
marrow.

Page 62 of 70
 Nitrogen Fixation
Genetic engineers have
developed plants that can
fix nitrogen directly from
N
oo
n

the atmosphere. Such

Ac
ad

plants need less fertilizer.

em
y

Page 63 of 70
SINGLE-CELL PROTEIN
Single-Cell Protein (SCP) refers
to the protein content
extracted from pure or mixed
cultures of algae, yeasts, fungi
or bacteria. For the production
N

of single-cell proteins, the

oo
n
Ac

microorganisms are grown in

ad
em

fermenters. These
y

microorganisms utilize a
variety of substrate like
agricultural wastes, industrial
wastes, natural gas like
methane etc.
Page 64 of 70
 Novel protein
Microorganisms grow very
vigorously and produce a
high yield of protein. The
N
oo
n

protein content produced

Ac
ad

by microorganisms is also
em
y

known as novel protein or

minifood.

Page 65 of 70
 SCP History
For a better management
of food shortage problems
the use of microbes as the
producers of single-cell
N
oo
n

proteins has been successful

Ac
ad

on experimental basis. This

em
y

technique was introduced

by Prof. Scrimshow of
Massachusetts Institute of
Technology.

Page 66 of 70
 Significance of SCP
 Scientist and food
technologists believe that
single-cell proteins will
substitute the other
N
oo
n

proteinrich foods in
Ac
ad

human and animal feeds.

em
y

 The microorganisms
grow very vigorously and
produce a high yield.

Page 67 of 70
 Significance of SCP
 When single-cell proteins
are produced by using
yeasts, the products also
contain high vitamin
content.
N
oo
n

 In the production of
Ac
ad

single cell proteins,

em
y

industrial wastes are used

as raw materials for
microorganisms. It helps
in controlling pollution.

Page 68 of 70
 Significance of SCP
 The use of single-cell
proteins has good
prospects in future
because they contain all
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oo
n

essential amino acids.

Ac
ad

 Moreover, the
em
y

production of single-cell
proteins is independent of
seasonal variations.

Page 69 of 70
 Examples of protein
productions
 It has been calculated that
50 kilogram of yeast
produces about 250 tons of
protein within 24 hours.
N

 Algae grown in ponds

oo
n
Ac

produce 20 tons (dry weight)

ad
em

of protein per acre/year. This

y

yield of protein is 10-15

times higher than soybeans
and 20-50 times higher
than corn.

Page 70 of 70