Unit 1

BIOTECHNOLOGY: AN OVERVIEW
OVERVIEW
Overview
Modern biotechnology has integrated several disciplines, varying from physics,
chemistry, mathematics, biology, engineering, economics, law and management.
The extraordinary developments occurring in biotechnology throughout the world
has lead to significant changes in world commerce, as well as in human healthcare
and agriculture sectors. Production of vaccines employing recombinant DNA
methods is one example of its potential application. Pharmacologically active
compounds from plants and marine organisms can now be isolated effectively
using biotechnology. Progress in agricultural biotechnology will make our society
less dependent on sustained harvest, which is often subjected to the vagaries of
weather or climate.
This chapter makes you familiar with the developments in biotechnology field and
its applications in health care and agriculture.

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 1 Chapter
The development of technologies that use living organisms or substances derived from them for the
benefit of mankind led to the establishment of field of biotechnology. Thus, biotechnology is now defined
as Any technological application that uses biological systems, living organisms or derivatives thereof to
make or modify products or processes for specific uses (Food and Agriculture Organization). Humans
have for centuries practiced biotechnology. They have used biotechnology to make bread, cheese, and
beers and wines, to breed strong and productive animals, and to increase the yield of crops by selecting
seeds from particularly desirable plants. The modern biotechnology is based on the fundamental
knowledge of biosciences and thus, truly interdisciplinary in nature (Fig. 1). It has already made its
presence felt in our daily life in many ways and now providing new opportunities for career
advancement.
1.1 Historical Perspectives
The making of curd, bread, wine, and beer originated in prehistoric kitchens and the process
leading to the development of these products came to be known as fermentation. Around 2000
BC, the Egyptians knew that when crushed dates were stored, a pleasantly intoxicating material
was produced at first but if the mixture was allowed to stand for a longer time, it turned sour to
yield vinegar, the strongest acid known to antiquity. By 1500 BC, the use of germinated cereals
(malt) for the preparation of beer from bread leaven (a mass of yeast) on a cereal dough and the
formation of wine from crushed grapes, were established as technical arts in Mesopotamia,
Palestine and Egypt. The ancients also observed that the formation of beer, wine or vinegar was
followed by changes that led to the liberation of noxious odors, thus showing putrefaction for
plant materials as compared to the more rapid decay of animal or human tissues. The practical
arts of preserving animal foods drying, smoking, curing, pickling in brine, and treatment with
granular salt -- were well developed in the prehistoric Near East and Europe. The well-known
mummification procedures in Egypt used the technique of dehydration with a mixture of salts,
largely sodium carbonate.
By18th century it was observed that fermentation could be classified into three groups on the
basis of final products:
1. Evolution of gas
2. Formation of alcohol, and
3. Production of acid
Antoine Lavoisier provided the chemical basis for the nature of alcoholic fermentation by using
analytical techniques for the quantitative estimation of carbon.

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1.1.1 Microorganisms as causative agents of fermentation
In the early 19th century, Nicolas Appert, a French manufacturer of confectionery who distilled
spirits and food products, described methods for preserving foods by putting them into tightly
closed vessels that were then heated in boiling water this marked the beginning of the canning
industry. Gay Lussac examined Apperts closed heated vessels and found that they lacked
oxygen. This led to the belief that oxygen was necessary for fermentation. The construction of
achromatic compound microscope demonstrated in 1837 that the agents of fermentation are
living organisms. Charles CagniardLatour in 1838 described the involvement of brewers yeast
in alcoholic fermentation based on: (i) its constant occurrence in fermentation, (ii) cessation of
fermentation under the conditions that killed yeast, such as boiling, treatment with arsenate, etc.
and (iii) it was evoked and increased by the process itself - a phenomenon that applies to living
organisms only.
In 1857, Louis Pasteur published his first report on formation of lactic acid from sugar through
fermentation. It was already known that lactic acid is produced from sugar and that addition of
chalk to fermentation mixture markedly increases the amount of lactic acid produced. Using a
microscope, Pasteur showed the presence of lactic yeast, which is the agent for the making of
curd and confirmed the presence of L-lactic acid using a polarimeter. In 1860, he presented a
detailed report on alcoholic fermentation and concluded:

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 (i) The act of fermentation is a phenomenon, which is unique, and very complex, as it can be a
phenomenon correlated with life, giving rise to multiple products such as succinic acid, glycerin
etc. all of which are necessary.
(ii) There is never any alcoholic fermentation without there being simultaneously the organization,
development, and multiplication of the globules which are already formed.
(iii) He held the same views on lactic fermentation, butyric fermentation, fermentation of tartaric acid
etc.
(iv) He did not comment on the chemical act of cleavage of the sugar.
(v) Fermentation appears to be a physiological phenomenon.
(vi) Fermentation is a consequence of anaerobic life.
Our ancestors have developed their own kitchen technologies with the help of fermenting
bacteria, like Lactobacillus, Leuconostoc, Lactococcus, Enterococcus, Pediococcus etc leading
to the development of a wide variety of yummy dishes that we relish today (Table 1). The
preparation of curd is a household recipe known to us for centuries. You too can prepare curd
using the following mothers recipe (Fig. 2):

MAKING OF THE CURD

Have you observed your mother making curd at home? Depending on the size of the family and average
consumption, she has arrived at her own quantitative approach. Assuming a family of four consumes
200 ml/person of curd, she takes 800-1000 ml of milk, warms/cools it to lukewarm temperature (~37oC).
Then she takes a teaspoon of the previous days curd and distributes it uniformly in one liter of milk by
stirring it. Assuming that it is a warm summer day, the curd is ready in about four hours.
Observations
The raw material, milk, has been converted completely into a semisolid product in four hours by
addition of only one teaspoon of curd.
The raw material, a white homogeneous slightly sweet liquid, has been converted into a
semisolid, sour tasting product.
There appears to be both a physical as well as a chemical change during the process.

The 19th century saw the growth of industries linked to fermentation, which gave rise to products like
wine, beer, and whisky. This period also saw the growth of canned food industry, which helped in the
preservation of food for a longer time and made them available during off-seasons. The fermentation
industry along with agricultural practices of animal husbandry and plant hybridization led to the shifting
of attention from agriculture to industry.
The 20th century saw man develop the ability to conquer several diseases thanks especially to the
antibiotic revolution in the 1940s. The antibiotic revolution demanded large-scale production to meet the
challenging demand of the health sector. This resulted in a shift from kitchen technologies to large-scale
manufacturing in an industrial sector. And this further led to close interaction between fermentation

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scientists and engineers from various disciplines of engineering, which gave rise to branches like
bioprocess engineering, food science engineering and technology, bio-expert system engineering and
artificial neural networks, validation engineering, metabolic engineering and environment management.

Fig. 2.. Making of the curd

TABLE 1. TRADITIONAL LACTIC-FERMENTED FOODS OF INDIA

Food Substrate Nature and use
Traditional fermented products of south India
Idli Rice-black gram Steamed, spongy cake; breakfast food
Dosa Rice-black gram Spongy pan cake, shallow-fried; staple food
Ambali Millet, rice Steamed cake; staple food
Traditional fermented products of north India
Bhallae Black gram Deep-fried patties; snack
Vadai Black gram Deep-fried; snack
Papad Black gram Circular wafers; snack
Wari Black gram Spongy cake; snack
Bhatura Wheat Flat deep-fried, leavened bread; snack
Nan Wheat Leavened flat backed bread; staple food

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 Jalebi Wheat Crispy, deep fried pretzel; sweet confectionery
Paneer Milk Soft milk-flavored cheese; fried curry
Dahi Milk Thick-gel, savory
Traditional fermented products of western regions of India
Dhokla Bengal gram Spongy cake; snack
Khaman Bengal gram Spongy cake; breakfast food
Rabadi Wheat/Pear-millet Cooked paste; staple food
/Barley-Buttermilk mixture
Srikhand Milk Concentrated sweetened, savory
Indigenous fermented products of eastern regions of India
Mishti dahi Milk Thick gel; sweet savory
Tari Date palm Sweet cloudy white alcoholic beverage
Traditional fermented products of the Himalayas
Gundruk Leafy vegetable Sun-dried, sour-acidic taste; soup/pickles
Sinki Radish tap root Sun-dried, sour-acidic taste; soup/pickles
Mesu Bamboo shot Sour-acidic, pickles
Khalpi Cucumber Sour, pickles
Chhurpi Milk Soft mass, cheese-like, mild sour; curry

1.2. Technology and applications of Biotechnology

The current excitement about modern biotechnology is due to the manner in which this science
has pushed organisms beyond their natural abilities by genetic engineering and hybridoma
technology. Nature has equipped every organism with the capacity to perform within an optimum
or balanced system. Some of the common Living systems with industrial applications are listed
in Table 2. Modern biotechnology has tinkered with the genetic material of the organism by
introducing foreign genes. The purpose is to push the organism to do things it never did before.
Ability to genetically manipulate organisms right from viruses to mammals led to the genomics
revolution touted as the third technological revolution following the industrial and computer
revolutions. For example, study of genomics has helped not only in determining the entire gene
sequence but also making fine-scale genetic maps of organisms. Bioremediation technologies
are used to clean our environment by removing toxic substances from contaminated soils and
ground water. Agricultural biotechnology has reduced our dependence on pesticides.
Manufacturing processes based on biotechnology has made it possible to produce paper and
chemicals with less energy, less pollution, and less waste. Biotechnology has now found
applications in virtually every sphere of human activity (Table 3). Different technologies used in
biotechnology and their applications are as follows
Table 2. Living systems with Industrial applications
Prokaryote
Eubacteria
Unicellular

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 Mycelial--Actinomycetes
(Streptomyces, Nocardia)
Archaea (extremophiles)
Eukaryote
Fungi
Yeast
Mold
Algae
Protozoa
Insect Cells
Plant cells or Tissues, organs
Animal cells, tissues and organs
Transgenic animal/Plant
1.2.1 Bioprocessing Technology
Bioprocess technology is the oldest in all the biotechnologies. Bioprocessing technology, uses
living cells or the molecular components to manufacture desired products. The living cells most
commonly used are one-celled microorganisms, such as yeast and bacteria; the biomolecular
components mostly used are enzymes, which are proteins that catalyze biochemical reactions.
Microbial fermentation,is a form of bioprocess technology has been used for thousands of
yearsunwittinglyto brew beer, make wine, leaven bread and pickle foods. Now a day
recombinant DNA technology coupled with microbial fermentation are used to manufacture a
wide range of biobased products including human insulin, the hepatitis B vaccine, the calf
enzyme used in cheese making, biodegradable plastics, and laundry detergent enzymes.
1.2.2 Cell Culture
Cell culture technology is the growing of cells outside of living organisms. Cell culture are of
many types.
Plant Cell Culture: An essential step in creating transgenic crops, plant cell culture also provides
us with an environmentally sound and economically feasible option for obtaining naturally
occurring products with therapeutic value. Plant cell culture is also an important source of
compounds used as flavors, colors and aromas by the food-processing industry.
Mammalian Cell Culture: Livestock breeding has used mammalian cell culture as an essential
tool for decades. Eggs and sperm, taken from genetically superior bulls and cows, are united in
the lab, and the resulting embryos are grown in culture before being implanted in surrogate cows
1.2.3 Recombinant DNA Technology
Recombinant DNA technology is viewed as the cornerstone of biotechnology. The term
recombinant DNA means the joining or recombining of two pieces of DNA from two different
sources. Genetic modification using recombinant DNA techniques allows us to move genes
whose functions are known. By making manipulations more precise and outcomes more certain,
the risk of producing organisms with unexpected traits are decreased. Recombinant DNA
technologies have various uses like research application, creation of genetically modified
variety of plants and animals.

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 1.2.4 Cloning
Cloning technology allows us to generate a population of genetically identical molecules, cells,
plants or animals. Because cloning technology can be used to produce molecules, cells, plants
and some animals, its applications are extraordinarily broad. Molecular or gene cloning is the
process of creating genetically identical DNA molecules. Molecular cloning is foundation of the
molecular biology and is a fundamental tool of biotechnology research, development and
commercialization. All applications in biotechnology, from drug discovery and development to
the production of transgenic crops, depend on gene cloning.
Animal cloning has helped us to rapidly incorporate improvements into livestock herds for more
than two decades and has been an important tool for scientific researchers. One of the
technique used in animal cloning is Somatic cell nuclear transfer (SCNT). It involves transfer of
the nucleus of a somatic cell taken from an adult female and transferred it to an egg cell from
which the nucleus had been removed. The resulting cell behaved like a freshly fertilized zygote,
which developes into an embryo.
1.2.5 Protein Engineering
Protein engineering technology involves improvement of existing proteins, such as enzymes,
antibodies and cell receptors, and to create proteins not found in nature. This technique is used
in conjunction with recombinant DNA techniques. Such engineered proteins are used in drug
development, food processing and industrial manufacturing.
1.2.6 Biosensors
Biosensor technology couples the knowledge of biology with microelectronics. A biosensor is
composed of a biological component, such as a cell, enzyme or antibody, linked to a tiny
transducera device powered by one system that then supplies power (usually in another form)
to a second system. Biosensors are detecting devices that rely on the specificity of cells and
molecules to identify and measure substances at extremely low concentrations. When the
substance of interest binds with the biological component, the transducer produces an electrical
or optical signal proportional to the concentration of the substance. Biosensors can be used for;
measurement of nutritional value, freshness and safety of food; location and measurement of
environmental pollutants.
1.2.7 Nanobiotechnology
The word nanotechnology derives from nanometer, which is one-thousandth of a micrometer
(micron), or the approximate size of a single molecule. Nanotechnology is the study, which
involves manipulation and manufacture of ultra-small structures and machines made of as few
as one molecule. Nanobiotechnology uses the knowledge of nanotechnology and biomolecules
of cell to produce desired technology. Some applications of nanobiotechnology includes; fast
diagnosis of disease, bio-nanostructures creation for getting functional molecules into cells,
improvement of specificity and timing of drug delivery.
Table 3. Applications of Biotechnology
Health Industry
Protein pharmaceuticals

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 Vaccine and Therapeutic agents
Diagnostics (Protein- or DNA)
Gene Therapy
Agriculture Industry
Biopesticides/ Biofertilizers
Crops tolerant to abiotic and biotic stresses-
Pharmaceutical production
Chemical Industry
Fermentation process to produce organic chemicals
Production of high purity chemicals
Use of energy efficient processes
Cleaning Industry
Use of enzymes as detergents like proteases
Textile Industry
Use of enzymes for finishing of fabrics
Use of genetically-modified cotton
Pulp and Paper Industry
Improvement of physical properties of fibers\
Biobleaching of pulp
The advent of the 21st century has witnessed the:
Mapping of genomes of many plant and animal species including humans,
Cloning of animals including many higher mammals,
Development of target-specific new molecules and drugs,
Stem cell therapy for genetic diseases and accident cases,
Tissue engineering for producing artificial tissues and organs ,
Creation of transgenic animals and plants for the production of recombinant bio-
pharmaceuticals,
Emergence of bioinformatics and computational biology in understanding the functioning of
cells and organs at genomic and protein levels etc.,
Application of in-silico experiments to understand pharmacology, toxicity etc.,
Development of automated gene and protein sequencing
Identification of proteins and peptides by mass spectrometry,
Evolution of nanotechnology and its application in nanomedicine, and
Growth of RNA interference biology
1.3 Global Market of Biotech Products
The biotechnology industry is also improving lives through its substantial economic impact.
Biotechnology has stimulated the creation and growth of small business, generated new jobs,
and encouraged agricultural and industrial innovation. In USA, the industry currently employs

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 over 150,000 people and invests ~ $10 billion a year on research and development (Table 4).
Table 4. U.S. biotechnology product sales forecast (Value in $ million)
Key sectors Base year Forecast year Forecast year
1998 2003 2008
Human therapeutics 9,120 16,100 27,000
Human diagnostics 2,100 3,100 4,300
Agriculture 420 1,000 2,300
Specialties 390 900 2,000
Non-medical diagnostics 270 400 600
Modern biotechnology products started coming into the market only in the late 1980s. Globally, the
share of biotechnology in the pharmaceutical market was less than 0.14% in 1985. By 2000 this share
grew to 20%. Today, about 60% of the biotechnology products in the market are healthcare products
(Table 5). About 21% of the biotechnology products go into agriculture and animal husbandry. The
commonly sought after agronomic traits for plants are given in Table 6. Given that there are some crucial
questions that have remained unanswered so far (for example, how a bacterially produced human
protein would acquire the same three-dimensional structure as found in the human system?), remain a
challenge and enigma for biotechnologists.
MAMMALIAN CELLS
Hybridomas, Myeloma Monoclonal Antibodies, recombinant antibodies
Chinese Hamster Ovary, Baby Hamster Interferon, tissue plasminogen
Kidney Cell lines activator (tPA), erythropoientin (EPO)
Human Kidney 293 Cell Lines Adenovirus for gene therapy
Human lung fibroblast MRC-5 Attenuated hepatitis C virus
Monkey Kidney epithelial cell Vero Inactivated polio virus
BACTERIA
Escherichia Coli Insulin, human growth Hormone, somatostatin,
interferon, Bovine growth hormone
YEAST
Saccharomyces cerevisiae Hepatitis B virus surface antigen (vaccine against
Hepatitis B)

Table 6. Agronomic traits from Biotechnology

Herbicide tolerance
Insect resistance
Disease resistance
Abiotic stress, e.g., Water, Temperature, Salt, Metals
Value added traits, e.g., Oil, Vitamins, Minerals
Nutritional quality, e.g., proteins, fats, carbohydrate; Plant properties, e.g., shelf-life, flavors

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 fragrance
Allergen reduction
Nitrogen fixation
Yield
Pharmaceutical production, e.g., Vaccines, Proteins
2001: GMO 130 MM acres (75% U.S.),
e.g., cotton, soybean, corn, canola, rice
________________________________________________________
Overall Benefits : Less cost / acre, More yield, Less pollution
1.4 Public Perception of Biotechnology
The public demands that biotechnology should work for the benefit of the society. This entails
major changes in the sciences and the social sciences. But these changes are not possible
without active societal participation. The participation of the society has become all the more
important because of the realization of the environmental consequences of science and
technology. A technology is democratic if it has been designed and chosen with democratic
participation. Participation in scientific research decision-making and policy-making, in general,
requires certain discretion and deliberative capabilities on the part of the citizens. Civil society
organizations (CSOs) have inherent advantages in handling information (on social, economic
and ecological variables and processes) and controlling decision processes. The debate about
the relative merits of representative participation and direct democratic participation of the
individual in environmental decisions remains unresolved. It is argued that representative
participation may be the solution for the time being, because detailed debates on technology or
the technology tribunals work only in contexts where there are literate and informed public and
a liberal democratic government. Therefore, the participation should begin at all levels of
decision-making so that the society could have better and more informed citizens.
While developments in science and technology are instrumental in most of our progress, some
genuine concerns have been raised regarding the long-term consequences on our health and
environment. In 1985, the UN assembly created the Brundtland Commission also called WCED
(World Commission on Environment and Development). The commission submitted a report on
the theme of sustainable development. It recognized that there is a real conflict between
meeting the needs and desires of 6 billion people living today and the possibility of satisfying the
ten billion people expected to inhabit the Earth by 2050!! The WCED defined Sustainable
Development as follows:
Sustainable development is development that meets the needs of the present without
compromising the ability of future generations to meet their own needs.
Definitions of sustainable development are abundant and are interpreted in varying manner by
different nations. In all these definitions and the way they have been implemented, science and
technology is invariably portrayed as a double-edged sword. A heated debate is taking place
globally over the issue of GM crops. This debate, which features science, economics, politics
and even religion, is taking place almost everywhere. Similar is the case with embryonic stem
cell research and its funding.

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 1.5 Biotechnology in India and Global Trends
1.5.1 Indian scenario
The Indian response to the biotechnology industry and its products has been very supportive
right from the early 1980s when the investments in the Department of Biotechnology (DBT)
(http://dbtindia.nic.in) and other research and development avenues were actively promoted.
The Government has set up a Technology Development Board to promote product development
with universities, industries and national institutes. Vision 2020 document has been prepared by
Technology Information, Forecasting and Assessment Council (TIFAC), which also includes
biotechnology. Indigenously developed Hepatitis B vaccine has been developed, manufactured
and marketed since 1998. There are nearly 30 companies devoted to development of modern
biotechnology products in India. Currently, Indias share in the global biopharmaceutical market
is just 1.5% and is valued at US $ 1.05 billion. By registering a high growth rate of ~32 per cent in
recent years, India is poised to become a major player in the coming years.
1.5.2 Global scenario
The present global biopharmaceutical industry is valued at US $ 71 billion. With the present
growth rate of 16 per cent, it is likely to cross $ 100 billion mark by the year 2010. USA promotes
biotechnology enterprise by gearing up the administration to develop policies, which will foster
biotechnology innovations as expeditiously and prudently as possible. There has been steady
increase in support for basic scientific research at the National Institutes of Health (NIH) and
other science agencies. The process of approving new medicines, making them available and
encouraging private-sector research investment and small business development are being
accelerated through new tax incentives and small business innovations research program. The
Government promotes intellectual property protection and open international markets for
biotechnology inventions and products; and developed public databases that enable scientists
to coordinate their efforts in an enterprise that has become one of the worlds finest examples of
partnership among university-based researchers, government and private industry. In addition,
the administration has strengthened efforts to improve science education and promote the
freedom of scientific enquiries. It has also provided guidelines to protect patients from the
misuse or abuse of sensitive medical information and provide Federal regulatory agencies with
sufficient resources to maintain sound, science-based review and regulation of biotechnology
products. It has also ensured that science-based regulatory programs worldwide promote
public safety, earn public confidence and guarantee fair and open international markets. The
European and the US biotechnology industries both have around 2000 companies, but the US
sector employs nearly twice as many people, spends around three times as much on research
and development, has twice the number of employees involved in research and development. It
also earns twice as much revenue compared to the European counterpart.
Review Questions
1. Do you agree that your grandmother unknowingly practiced a technique of biotechnology?
Describe the technique.
2. What are the two instruments Louis Pasteur used to show the presence of lactic yeast and L-
lactic acid?

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3. Name four traditional fermented foods of India.
4. Name some of the living systems that are being used for the industrial applications.
5. List the important applications of biotechnology in health care.
6. List the important applications of biotechnology in agriculture.
7. Which was the first recombinant DNA-based product produced and marketed in India?
Further Reading
1. DNA Science: A First Course, 2nd Edition, by David A. Micklos and Greg A. Feyer (Cold Spring
Harbor Laboratory Press, New York), 2003.
2. Milestones in Biotechnology, by J. Davies and W.S. Reznikoff (Butterworth-Heinemann,
Boston), 1992.
3. Textbook of Biotechnology, 3rd Edition, H. K. Das (ed.) (Wiley India Pvt Ltd., New Delhi),
2007.
4. Recombinant DNA and Biotechnology A Guide for Students, by H. Kreuzer and A. Massey
(ASM Press, Washington), 2001.

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