Introduction to Biotechnology XYAM PANT

INTRODUCTION TO BIOTECHNOLOGY
Modern biology is the most diversified of all the natural sciences, with numerous sub-disciplines like
microbiology, plant and animal anatomy, biochemistry, immunology, cell biology, molecular biology, plant
and animal physiology, morphogenesis, systematics, ecology, genetics, biotechnology and many others.

Biotechnology = Biology + Technology

The principles of the life sciences (biology) and engineering/technology for manufacturing processes
encompass biotechnology. The biological sciences provide principles/theories. The engineering sciences
mainly provide technology so as to change or modify the bioprocesses/organisms, to design the equipments,
to scale-up, purify, separate and recover the products or services. These engineering fundamentals required
to translate the discoveries of biotechnology into commercial products, thereby making biotechnology to
work, are collectively known as bioprocess engineering (National Research Council 1992). The principles
and practices of bioprocess engineering are invariably based on the biological sciences and the biotechnology
provides a foundation for the engineering of bioprocesses for production of various products. The bioprocess
engineering/technology is therefore required for operating the biotechnology. The bioprocess engineering
applies the technology in biotechnology for the followings:
1.For exploiting biochemical processes
2.For designing the equipment and facilities to manufacture the bioproducts.
3.For identification of genes, and the products that result from them
4.For scaling-up, separate, purify and recover the products,
5.For application of molecular biology to obtain transformed microorganisms, cells or animals and their
products.
6. For the development of biological sensors coupled to microprocessors or computers for process control and
monitoring of biological systems

There is no universal definition of biotechnology; however, some widely accepted definitions of

biotechnology are as under:
1. The application of biological organisms, systems or processes to manufacturing and service industries.
2. The integrated use of biochemistry, microbiology and engineering sciences in order to achieve
technological (industrial) application capabilities of microorganisms, cultured tissue cells and parts
thereof.
3. A technology using biological phenomena for copying and manufacturing various kinds of useful
substances.
4. The application of scientific and engineering principles to the processing of materials by biological
agents to provide goods and services.
5. The science of the production processes based on the action of microorganisms and their active
components and of production processes involving the use of cells and tissues from higher organisms.
Medical technology, agriculture and traditional crop breeding are not generally regarded as
biotechnology.
6. Really no more than a name given to a set of techniques and processes.
7. The use of living organisms and their components in agriculture, food production and other industrial
processes.
8. The deciphering and use of biological knowledge.
9. The application of our knowledge and understanding of biology to meet practical needs.
10. The European Federation of Biotechnology (EFB) defines biotechnology as ‘the integration of
natural sciences and organisms, cells, parts thereof, and molecular analogues for products and
services’. The EFB definition is applicable to both ‘traditional or old’ and ‘new or modern’
biotechnology.

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Introduction to Biotechnology XYAM PANT

HISTORY OF BIOTECHNOLOGY
Biotechnology is not a new science but something
which represents a developing and expanding
series of technologies dating back thousands of
years, when humans first began unknowingly to
use microbes to produce foods and beverages like
bread and beer and to modify plants and animals
through progressive selection for desired traits.
From historical perspective, biotechnology can be
categorized as- traditional or kitchen
biotechnology and new or modern
biotechnology. This categorization has been based
on the microorganisms exploited (either
genetically modified or not) and products
recovered.
As a traditional technology, biotechnology
encompasses many traditional processes,
especially for the production of foods and
beverages, such as brewing, baking, wine making,
production of cheese and oriental foods such as soy
sauce and tempeh, where the use of
microorganisms has been developed somewhat
empirically over countless years. Since the
traditional biotechnology involved mainly the
production of foods and beverages, it is also

known as kitchen technology/prior

biotechnology.
The era of new biotechnology started after the
discovery of microorganisms and their use in
various industries. New Biotechnology is therefore
considered to have been evolved through the
industrial microbiology. (The branch of
microbiology which is dedicated to the use of
microorganisms in various industries is called
industrial microbiology).
However, the new biotechnology revolution
began in the 1970s and early 1980s with the
development of ‘genetic engineering’ and ‘cell
fusion’ to alter precisely the genetic constitution
of living organisms. New biotechnology is defined
as the science that enables
manipulation/alteration of the cell’s genetic
constitution through recombinant DNA techniques
and cell fusion to produce such organisms that
have the improved or desirable characteristics or
that are capable of producing useful products.
Bioprocess engineers and scientists have
collectively combined relevant biological and

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engineering knowledge and expertise to
change the genetic makeup of microbial,
plant, and animal cells for the productive
use. Thus, the directed manipulation of
genes distinguishes the new biotechnology
from prior/traditional biotechnology
Biological molecules/products, for which
there is no other means of industrial
production, are generated through new
biotechnology. Existing industrial
microorganisms are systematically altered
(i.e. engineered) to enhance their function
and to produce useful products in new ways.
The new biotechnology through genetic
engineering has fundamentally expanded the
utility of biological systems.
The new biotechnology had a profound
impact on almost all areas of traditional
biotechnology and also permitted
breakthroughs in medicine, agriculture and
industrial manufacture that would be
impossible by traditional biotechnology.
Some of the most exciting advances were
the development of new pharmaceutical
drugs and therapies, production of healthier

foods, selective pesticides, and innovative

environmental technologies. The modern
biotechnology, thus, established a huge and
expanding market, that in monetary terms
represents financial profits. The modern
biotechnology has revolutionized human and
animal food production, alternative energy
sources, waste recycling, pollution control,
agriculture, aquaculture, forestry, many
aspects of medicine, veterinary sciences,
pharmaceutics and many other fields. The new
techniques had also revolutionized many
aspects of medicine, veterinary sciences, and
pharmaceutics. The recent mapping of the
human genome, recognised as one of the most
significant breakthroughs in human history
had only been possible because of
biotechnology.

NOTE: Developments subsequent to 1995 may

be found in:
Ladisch MR (2005). Biotechnology (Bioprocess
Engineering)
http// qap2.onlinelibrary.wiley.com

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Introduction to Biotechnology XYAM PANT
BIOTECHNOLOGY: AN INTERDISCIPLINARY SCIENCE
Biotechnology is an interdisciplinary science. An ‘interdisciplinary’
science takes ideas and techniques from different disciplines but has its
own characteristic concepts and methodologies. An interdisciplinary
science opens up a entirely new spectrum of investigations. Many
aspects of biotechnology have arisen through the interaction between
various parts of biology and engineering. Unlike a single scientific
discipline, biotechnology can utilise techniques derived from a wide
range of relevant fields like chemistry, microbiology, biochemistry,
chemical engineering, computer science, molecular biology, cell
biology, immunology, protein engineering, enzymology, breeding
techniques, bioprocess technologies, etc (see Fig.). The applications of
biotechnology essentially rest upon each of the contributing disciplines
and biotechnologists, therefore, work in a close cooperation with
experts from other related fields such as medicine, nutrition, the
pharmaceutical and chemical industries, environmental protection, waste process technology, etc. One of the
major advantages of such interdisciplinary science is they help to understand the potential as well as the
limitations of the other disciplines and help in development of innovative ideas to solve the problems.
Besides being an interdisciplinary science, biotechnology is a demanding industry. The main types of
companies involved with biotechnology can be placed in seven categories (see table). Biotechnology is not
itself a product or range of products rather it is a combination of enabling technologies that have significant
application in many industrial sectors. It is a technology in search of new applications, with many benefits
still to unfold.

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BIOTECHNOLOGY: A THREE-COMPONENT CENTRAL CORE
Many biotechnological processes may be considered as having a three component central core:
1. Component I: Isolation, screening and improvement of biological agents i.e. obtaining the best biological
catalyst/agents (eg. Microbes/plant or animal cells) for a specific function or process
2. Component II: To construct optimum environment for biological agents/catalysts eg. by construction of
bioreactors and technical operation
3. Component III: Product recovery/downstream processing

AREA/SCOPE/APPLICATION OF BIOTECHNOLOGY
The scope of biotechnology is broad and deep. Biotechnology encompasses the use of chemicals to modify
the behavior of biological systems, the genetic modification of organisms to confer new traits, insertion of
foreign DNA into the organisms to produce the desirable characteristics or to compensate for genes whose
absence cause life–threatening conditions, bioinformatics, etc.

1. Food and Beverage Biotechnology:

This branch of biotechnology encompasses a wide range of
options for improved quality, nutrition, safety and preservation
of foods and deals with the production of industrially valuable
food products and beverages. The impact of biotechnology on
the food and beverage industries can be:
-agronomic, i.e. increased plant and animal yields, extended
growth range and environments from which the farmers will
mainly benefit;
-non-agronomic, i.e. improving plants and microorganisms to
provide benefits to the food producer, retailer or consumer. eg
by lowering the costs of manufacturing operations; keep fresh
longer; improve texture and taste .
The use of fermentation to produce fermented foods has a very long history. Microbial fermentation is
essential to production of wine, beer, bologna, butter milk, cheeses, kefir, olives, salami, sauerkraut,
bread, sweeteners, soy sauce, organic acids, pickles, vineger, amino acids, enzymes, mushroom,
vitamins and many more (see table). The use of microorganisms in production of foods has other advantages
as well:
i) Metabolic end products produced by the microorganisms flavor fermented foods. For example, mold-
ripened cheeses owe their distinctive flavors to the mixture of aldehydes, ketones, and short-chain fatty acids
produced by the fungi.
ii) These organisms are also of particular importance in the food fermentation industry because they produce
bacteriocins that inhibit the growth of undesirable organisms that cause food spoilage and the multiplication
of food borne pathogens. Eg. nisin produced by Lactobacillus lactis and monesin Streptomyces
cinnamonensis.
iii) Microorganisms are an excellent source of single-cell protein (SCP) because of their rapid growth rate,
their ability to use very inexpensive raw materials and the uniquely high efficiency to transform these raw
materials to protein. The term single-cell protein, or SCP, describes the protein-rich cell mass derived from
microorganisms grown on a large scale for either animal or human consumption. SCP has a high content of
protein containing all the essential amino acids. Organisms employed for the production of SCP: Fusarium,
Methylophylus, Chlorella, etc.
The new biotechnology techniques especially create possibilities of rapidly improving the quantity and quality
of food available.

2. Medical Biotechnology:
The health care sector has maximally utilized the biotechnology for the manufacturing products with
therapeutic value. Of all the biotechnology derived products, ~35% are considered to be of medical use and
for health care. The major aspects of medical biotechnology are outlined as under:

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A-Production of therapeutic
proteins and enzymes
One of the most dramatic and
immediate impacts of genetic
engineering/new biotechnology
was the production of large
amounts of hormones and
several other proteins encoded
by human genes by bacteria
through fermentation. (See
table 2.1)
i. Protein Hormones:
Insulin (was the first
genetically engineered
therapeutic agent produced by
genetically engineered E. coli
to be approved for clinical use
in humans since 1982) and
Human growth hormone,
(hGH), have been produced in
genetically engineered E. coli
in large amounts, at relatively little cost.
ii. Enzymes
Human tissue plasminogen
activator (tPA), a proteolytic
enzyme with a property for
degrading the fibrin clots, is
a therapeutic protein made
available in large amounts as
a consequence of
recombinant DNA
technology.
Production of chymosin
(rennin) in genetically
modified E. coli and
prochymosin produced in
genetically modified yeasts.
Other enzymes produced in
genetically engineered
organisms include
urokinase, superoxide
dismutase, etc
iii. Other proteins
HBsAg (Surface antigen of
Hepatitis B virus) have been
successfully produced in
genetically modified yeasts
Interferons, Interleukins,
serum albumin, etc are also
produced through
genetically engineered
organisms (see table 2.1)

B- Vaccine Production
Attenuated, Polypeptide,
Recombinant (eg. For

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Hepatitis B) and DNA vaccines
C-Production of secondary metabolites (Antibiotics)
A number of secondary metabolites produced through fermentation have been proved to be invaluable as
antibiotics, anticancer drugs, immunosuppressants, etc. Genetically modified microorganisms have been used
to produce such compounds in large amounts. Among these, antibiotics are the secondary metabolites
considered among the most important to human therapeutics, because of their selective toxicity for bacteria,
fungi, or protozoa. (See table 2.2)
D- Gene therapy
Gene therapy is a treatment strategy that involves the introduction of genes or genetic material into human
cells to alleviate or eliminate disease. The aim of gene therapy is to replace or repress defective genes that
cause the disease.
E- In-vitro fertilisation
F- Production of monoclonal antibodies and serological kits

3. Agriculture Biotechnology
Biotechnology is making new grounds in the agriculture area. Some approaches of agriculture biotechnology
are:
A- Biotechnology has become a useful tool for the production of biofertilisers and for the production of
biopesticides.
B- Biotechnology made the tissue culture techniques possible which is widely used these days for mass
propagation of plants.
C-Production of plant compounds through fermentation
Through cell culture techniques industrial production of essential oils, alkaloids, pigments, etc have been
made possible.
D-Control of pathogenic bacteria, fungi, and parasitic nematodes
The cell walls of many plant pests, such as insects and fungi, contain chitin (poly-N-acetylglucosamine) as a
major structural component. Genes encoding chitinases from several different soil bacteria have been cloned
into Pseudomonas fluorescens, an efficient colonizer of plant roots. The plant pathogens in the rhizospheric
zone are thus killed by the chitinase produced by the recombinant Pseudomonas fluorescens that has
colonised plant roots. However, the effectiveness of these recombinant strains in controlling fungal disease is
not yet known.

E- Transgenic plants
i. Resistance to insect pests
Bt plants have been produced by incorporating insecticidal toxin producing Bt genes of Bacillus thuringiensis
into the desired plants.
ii. Ability to grow in harsh environments
Several cold resistant, dessication resistant and disease resistant crop varieties have been produced. Plants that
have the ability to withstand high moisture or high salt concentrations; and resistance to iron deficiency in
very alkaline soils are also produced.
iii. Herbicide tolerance
Crop plants have been modified to become resistant to particular herbicides. These herbicides act as selective
weed killers when applied to a weed-infested field containing such genetically modified plants.
iv. Crop varieties with improved or new traits
-The crop varieties with increased productivity are produced.
-Cell culture and protoplast fusion techniques have resulted in hybrid and cybrid plants through which are not
possible through conventional hybridization techniques.
-Transgenic ‘golden rice’ with vitamin A producing genes has been produced.
-Plant biotechnology has resulted in the production of encapsulated seeds, soma clonal variants and transfer of
nod/nif genes to those plants that lack it (nod/nif genes are required for nitrogen fixation).

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4. Animal biotechnology
Animal agriculture in the form of cattle, pigs, sheep, poultry and fish represents a major aspect of food
production worldwide. Many of these animals are produced for their meat, milk and eggs and contribute to
human nutrition. In the modern world animal production is highly intensified and bio-technologically driven
to meet increasing demand for animal products.
A- The advent of recombinant DNA technology and its
application to livestock/animals has resulted in transgenic
animals with the improvements in meat production from a
wide range of farmed animals, including fish, improved milk
yields and quality, and disease-free animals (see table 10.8).
B- Molecular farming, a new concept has been introduced in
which therapeutic drugs are produced in farm animals.
Thecommercially feasible projects are the use of certain
lactating animals such as sheep, pigs, rabbits and cows to
produce novel secretions of human proteins in their milk which
can then be extracted and used pharmaceutically.

5. Environment Biotechnology
In the recent years the transformation of domestic wastes and industrial effluents, removal of xenobiotic
chemicals and treatment of sewage have drawn much attention for the protection of environment and for the
reduction of pollution. To combat these problems genetically engineered microorganisms have been
developed that can degrade the xenobiotic chemicals into non toxic forms that would otherwise remain
undegraded.
A- Wastewater Treatment
The volume of water being contaminated and the need to reclaim wastewater (sewage, industrial effluents,
agricultural runoff, storm water and urban runoff) are increasing with the growth in population and industrial
use. Treatment of wastewater is essential to prevent contamination of drinking water and the entry of
pathogens and contaminants into the food chain. Strains of bacteria, cyanobacteria and fungi have been
genetically modified which could be used for treatment of municipal and domestic sewage and industrial
discharge into non toxic forms and renew them as source of energy. eg. bacteria of Zoogloea species
(genetically modified ) play an important role in sewage treatment.

B- Bioremediation
Bioremediation is a process to clean up pollutants dispersed in the environment by the activities of living
organisms. Bioremediation organic pollutants to harmless inorganic products and is generally inexpensive
and causes little disturbance to the environment. Genetically modified microbes, have the capacity to degrade
a wide spectrum of environmental pollutants.
- For the cleanup of massive oil spills genrtically modified Pseudomonas putida (aka multi-plasmid
hydrocarbon-degrading Pseudomonas) has been used.

- Highly chlorinated aliphatic and aromatic compounds have potential adverse environmental and health
impacts and are resistant to degradation. These compounds are either fully or partially degraded by the
combined activities of various genetically modified bacteria such as Dehelobacter restrictus, Geobacter
lovleyi, Dehalococcoides sp., Syntrophus and Sytrophobacter.

- Subsurface/underground bioremediation of radioactive components such as uranium(U), strontium(Sr),

plutonium(Pu), cesium (Cs), and technetium (Tc) to prevent leaching and widespread contamination of
groundwateris increasing attention . Eg. Uranyl ion immobilization through subsurface reduction of U(VI) to
U(IV) by Geobacter Species, immobilization of Uranyl Ion mediated by Pseudomonas aeruginosa
polyphosphate metabolism.
C- Biomining (heavy metal extraction using microorganisms) utilizes naturally occurring prokaryotic
communities. Here, microorganisms are used to leach metals, principally copper but also nickel and zinc,
from low-grade sulfide- and/or iron-containing ores. The process exploits the energy metabolism of various
acidophilic chemolithoautotrophs that utilize inorganic compounds as energy sources and CO2 as the source
of carbon.
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D- Biotechnology has helped the bio-industries in bio-ethanol production, bio-hydrogen production and SCP
production from many waste products thereby appropriately managing them.

Alternatively The Main Areas Of Application Of Biotechnology Can Also Be Classified As

Bioprocess technology
Historically, the most important area of biotechnology, namely brewing, antibiotics, mammalian cell culture, etc.; extensive
development in progress with new products envisaged, namely polysaccharides, medically important drugs, solvents, protein-
enhanced foods. Novel fermenter designs to optimize productivity.
Enzyme technology
Used for the catalysis of extremely specific chemical reactions; immobilization of enzymes; to create specific molecular
converters (bioreactors). Products formed include L-amino acids, high fructose syrup, semi synthetic penicillins, starch and
cellulose hydrolysis, etc. Enzyme probes for bioassays.
Waste technology
Long historical importance but more emphasis now being made to couple these processes with the conservation and recycling
of resources; foods and fertilizers, biological fuels.
Environmental technology
Great scope exists for the application of biotechnological concepts for solving many environmental problems – pollution
control, removing toxic wastes; recovery of metals from mining wastes and low-grade ores.
Renewable resources technology
The use of renewable energy sources, in particular lignocellulose, to generate new sources of chemical raw materials and
energy – ethanol, methane and hydrogen. Total utilisation of plant and animal material. Clean technology, sustainable
technology.
Plant and animal agriculture
Genetically engineered plants for improved nutrition, disease resistance, keeping quality; improved yields and stress tolerance
will become increasingly commercially available. Improved productivity, etc., for animal farming. Improved food quality,
flavour, taste and microbial safety.
Health care
New drugs and better treatment for delivering medicines to diseased parts. Improved disease diagnosis, understanding of the
human genome – genomics and proteomics, information technology.

RISKS AND HAZARDS OF BIOTECNOLOGY

(TU exam)
In all biotechnology processes, safety is of paramount importance. When properly practiced, biotechnology is safe
and the benefits deriving from biotechnological innovations will surely lead to major improvements in the health
and well-being of the world’s population. However, biotechnology must always be subjected to sound regulations
for its successful application. The potential risks of biotechnology are manageable, and regulations have been
constructed for that management. The major hazards and risks associated with biotechnology can be summarized as:

1. Human health
The risks to human health are of major concern. Genetically modified foods may develop food allergy. The genetically
modified food may pose risk to ecology and environment as well. Genetically modified organisms (GMO) may spread
antibiotic resistance and pose a threat by developing new antigenic characters. Eg Penicillinase producing organisms
like Staphylococcus can transfer resistant gene to the organisms that lack it. Such genetic transfer can occur in the
biotechnology laboratories as well that provide a favourable environment.

2. Environment
There are threats to sensitive environmental issues. eg. Development of superweed due to horizontal or vertical gene
transfer. The development of resistance by pests against Bt plants is gaining concern. It is also possible that GMOs may
produce substances that stimulate plant pathogens instead of killing them.
Chimeric/recombinant/hybrid DNA may be taken up by the other organisms. Thus, they get genetically modified and
the natural characteristics of the organisms may alter which may cause disturbances in food chain and dis-balance
ecosystems. Besides, transgenic instability and unpredictable gene expression are potential hazard to environment.

3. Socio-economic impacts
a) Competition for land: For any biotechonology industry to produce the products it requires land. There may be
possible competition among various sectors for land acquisition.
b) Unemployment: Using biotechnological procedures it requires less employees to produce products than the
conventional industries thereby increasing unemployment.
c) Inflation: The use of biotechnology increases the cost of agriculture which may cause unpredictable inflation of the
food products.
d) Small scale farmers cannot compete with the improved varieties.

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4. Danger of new form of life
Constant working on gene and its manipulation may result in new organisms.

5. Extinction of the species

With the biotechnological applications the farmers choose improved varieties of crops. In due course of time, wild
varieties may extinct due to their un-selection. The wild varieties may not be available in the future if needed. The
misuse of biotechnology may cause a lot of mischief when trans-national companies engage in ruthless pursuit of profit.
This may lead to genetic erosion which brings a situation where base genetic material is available to only a few
multinational companies.

6. Release of genetically manipulated organisms into the environment

Recombinant microorganisms are now knowingly released into the environment e.g. for biological control, as
inoculants in agriculture, as live vaccines, for bioremediation, as baker’s and brewer’s yeast, etc. In addition, increasing
numbers of recombinant plants are taken out from research laboratories to the fields and greenhouses of the farmer and
large commercial horticulture grower. The released genetically modified microorganisms (for bioremediation,
biological control or as live vaccine) could transfer the hybrid DNA in the new microorganism altering its metabolic
activity in unpredictable and undesirable ways or may increased pathogenicity of normal microorganisms found in
environment. Similarly, there have been fears that transgenic crops move out of the field of cultivation and become
weeds. There is a possibility of gene transfer from herbicide and pest resistance transgenic plants to compatible wild
relatives making them ‘superweeds’.

7. Bioterrorism and biowarfare

There is a growing worldwide concern that pathogenic microorganisms or their products such as toxins may be used in
acts of urban terrorism. There is also potential with bio-agents to target farm animals and crops, which could cause
devastating economic effects. The production of most of the hazardous microorganisms and their harmful products is
relatively straight forward with suitable fermentation facilities.
Potential biological agents that have potential to be used as bio-weapons are assigned to three categories:
category A agents include the most serious illness– smallpox, anthrax, plague, botulism, tularaemia and viral
haemorrhagic fevers such as Ebola;
category B agents have a similar potential for large-scale dissemination but generally cause less serious illnesses –
typhus, brucellosis and food poisoning agents such as Salmonella and E. coli 0157;
category C agents include novel infectious diseases which could emerge as future threats.

8. Social conflict and ethical issues

The new biotechnology has provoked a variety of views within the public regarding biotechnology products. The
opinions are influenced by nationality, religion, ethics, morality and knowledge. In many societies the biotechnological
products have been unaccepted due social and ethical issues.
Some of the main ethical concerns relating to the food use of certain transgenic organisms are:
(a) Transfer of human genes to food animals (e.g. transfer of human gene for factor IX, a protein involved in blood
clotting, into sheep)
(b) Transfer of genes from animals whose flesh is forbidden for use as food by certain religious groups into animals
which they normally eat (e.g. pig genes into sheep would offend Jews and Muslims);
(c) Transfer of animal genes into food plants which may be of particular concern to some vegetarians
(d) Use of organisms containing human genes as animal feed (e.g. yeast modified to produce human proteins of
pharmaceutical value and the spent yeast then used as animal feed).
(e) Lard oil (an antifoaming agent, produced by application of biotechnology), is unaccepted by Muslims

9. Problems of biologically active biotechnology products

A care must be taken to prevent the indiscriminate dispersal of biologically active biotechnology products; vaccines and
antibiotics are obvious examples. Contaminants in such products may produce toxic molecules leading to poisoning or
allergenic reactions. Over-use of antibiotics in agriculture could lead to carry-over into human foods, resulting in
possible development of antibiotic resistance in human disease organisms.

(NOTE THAT: In the context of health and safety, ‘hazard’ can be defined as a substance, object or situation with a potential for
an accident or damage, and ‘risk’ is the likelihood that hazard will occur.)

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