MODULE 3

ENTREPRENEURSHIP OPPORTUNITY IN
INDUSTRIAL BIOTECHNOLOGY
• Business opportunity, Essential requirement, marketing
strategies, schemes, challenges and scope - with case
study-
– Pollution monitoring
– Bioremediation for Industrial pollutants, Pesticides, Herbicides etc.
– Integrated compost production - microbe enriched compost
– Biopesticide / insecticide production.
– Fermented products-probiotic and prebiotics.
– Stem cell production, Stem cell bank, contract research.
– Production of monoclonal/polyclonal antibodies
– Single cell protein and
– Secondary metabolite production
• Contact research in microbial genomics
 BUSINESS OPPORTUNITIES
i. Pollution monitoring

• In municipalities where the environmental agencies

impose regulations to reduce and control
emissions, foreign companies can find business
opportunities and present new technologies.
• Steel or cement production are examples of two such
areas.
• Furthermore, the large number of sugar mills produce
ethanol burn bagasse for cogeneration and are in need
of exhaust treatment technologies.
• Purchasing within the industry is done through bidding
processes and decision makers are usually accessible.

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• IBM is also looking at renewable power generation
companies.
– They already have 30 solar farms in China using their
forecasting technology to help predict the availability of sunlight
• Microsoft created a website called Urban Air and a
smart-phone app that gives a 48-hour air quality forecast
for cities across China
• One start-up called Air Visual, is a pollution monitoring
company based in Beijing, and they are already giving
the two tech giants a run for their money.
– Air Visual uses a smart-phone app and their website to give
users a free, three-day forecast for countries around the world

3
• Delhi-based Kaiterra creates highly-accurate monitors
that measure a variety of air pollutants, for consumers,
businesses and industrial use.
– These devices allow users to better understand their own air
quality, and creates maps of outdoor air quality.
• Delhi-based start-up PerSapien includes scientists,
doctors, engineers from Stanford University, AIIMS, IIT
and other international Universities, who deeply felt the
need to act quickly to save human lives.
• To empower people with the 'Right to Breath' pure air,
they invented 'Airlens'.
– It is a novel, nasal-air-purification device that effectively filters out
harmful pollutants like PM 2.5, PM 10, harmful gases etc. from
the inhaling air.
– The second component of Airlens is Airlens Data' app which
provides personal-level (hyperlocal) Air Quality Index (AQI) with
protection tracking feature.

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ii. Bioremediation for Industrial pollutants,
Pesticides, Herbicides

• Data mining of the current data to provide more insights

into the mechanisms of bioremediation.
• Numerical modeling and numerical simulation, which
requires development of novel algorithms.
• Identification of novel biomarkers.
• Those markers might be used for diagnostic of
bioremediation operations.
• The biomarkers could be functional genes, important
enzymes, and indicator metabolites formed during the
bioremediation reactions.

5
• Integration of data generated by functional genomics,
proteomics, transcriptomics, and metabolomic data,
provides a more complete picture of a microbial
bioremediation system and achieve a detailed
understanding of the physiological state of microbes used
for bioremediation purposes.
• International market for remediation sector is estimated to
be in range of US$30-35 billion
• Worldwide demand for bioremediation and
phytoremediation cleanup technologies is valued at $1.5
billion per annum
• Eg. Canadian environment industry has annual sales of
over $20 billion and contributes to 2.2% to Canada’s GDP
– Remediation as part of solid and hazardous waste management
sector comprises second largest component (24%) of Canada’s
environment industry

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iii. Integrated compost production – microbe
enriched compost
• A unit of capacity to process 20-30 tonnes of waste per
day can be set up within a budget of INR 50 Lakhs.
• The compost is also made as a by-product of a biogas
plant.
• The biogas plant converts the organic waste into bio gas
through anaerobic digestion and produces a slurry,
which can be dried and used as compost.

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iv. Bio-pesticide / Insecticide Production

• Organic Food business is USD 40 Billion and growing

• Therefore demand for non toxic solutions for crop
production and crop protection will increase.
• Bio-Pesticides is basically Non GM business
• Grown at a Compound Annual Growth Rate (CAGR) of
20.2% since 2010-2020
• Scope of Current market: USD 23.92 million (2015) for
bio-pesticides
• Represent only 4.2% of the overall pesticide market
• Over 150 Bio pesticide producing companies
• 15 types of bio pesticides out of 227 pesticides
registered

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v. Fermented products-probiotic
and prebiotics.
• The global probiotics industry was estimated at $19.6
billion in 2013.
• The global market for probiotic foods and drinks is
pegged at $32.6 billion, with Europe and Asia
contributing to 42 per cent and 30 per cent of the total
revenues.
• The industry estimates suggest the growth of the
industry at a CAGR of 12.6 per cent, with the increase
said to be for a span of four years beginning 2009.
• Introduced in India in 2007, its current stature stands at
$10 million with growth figures indicating an 8 to 10 per
cent increase for the past 3-4 years.
• The Indian probiotic industry stands at about Rs 50 crore
and is growing at 20 per cent. 9
vi. Stem cell production, stem cell bank
• The global market for stem cell products and services is
already valued at approximately €20bn and is forecast to
grow almost threefold by 2010 to €54bn.
• The global stem cell manufacturing market is projected
to reach USD 14.61 Billion by 2023 from USD 10.19
Billion in 2017, at a CAGR of 6.3 % during 2018–2023.

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vii. Production of monoclonal/polyclonal
antibodies
• The global antibody production market size was valued
at USD 7.4 billion in 2016 and is projected to grow at
13.5%
• The market for monoclonal Antibodies is projected to
increase at a compound annual growth rate of 20.8%
through 2010
• Within Big Pharma, mAbs are excepted to contribute
additional revenue of roughly $17.9 billion in 2010
• There are 418 biotechnology products either in human
clinical trials or under review by the US Food and Drug
Administration.
• mAbs account for 38% or 160 of these products and
recombinant proteins and hormones account for 10% or
43 products 11
viii. Single Cell Protein
ix. Secondary Metabolite Production
• Algae products such as Spirulina and Chlorella, have
significant benefits as potential single cell protein
sources.
• But the market value of these products is not very high;
– Spirulina was sold at a price of US $20/kg in 2010 and
– Chlorella at a price of US $44/kg in 2010.
• Microalgae extracts are also employed in making anti-
aging cream, refreshing or regenerant care products,
emollient and as an anti-irritant in peelers and are also
present in sun protection and hair care products.

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• Algae can also yield various carotenoids such as
Astaxanthin, Beta carotene, Lutein and many more,
which has potential applications in nutraceutical and
pharmaceutical industries.
• Beta-carotene has the largest share of the carotenoids
market.
• Valued at US$247 million in 2007, this segment is
expected to be worth US$285 million by 2015, a CAGR
of 1.8%.

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 ESSENTIAL REQUIREMENTS
i. Pollution Monitoring

• The first step in almost any air quality related regulatory

analysis is to define pollutant emissions from a source or
facility by completing an accurate emissions inventory.
– A properly estimated inventory will determine which regulations
apply and what levels of pollution control should be applied.
– Most facilities subject to air permit requirements must conduct
some kind of emission monitoring to judge compliance with
emissions limits.
• Air pollution instruments play an important role in the
determination of emission factors and the appropriate
pollutant emitting rates.

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• Also, air quality regulations in many countries require
either ambient air quality monitoring both before and
after a facility is constructed or a demonstration of
meeting air quality standards through modeling.
• All these activities involve measurement of
environmental related data.

ii. Bioremediation for Industrial pollutants, Pesticides,

Herbicides
• In-situ Methods
– Natural attenuation is connected with the degradation activities
of indigenous microorganisms.
• This method avoids damaging the habitat, allows ecosystem revert to its
original condition and enables detoxification of toxic compounds

15
– Bioaugmentation process, in which the specific degraders are
introduced into the soil.
• This method is applied when the indigenous microflora are unable to break
down pollutants, or when the population of microorganisms capable of
degrading contaminants is not sufficiently large.
• To make the process of bioaugmentation successful, microorganisms
introduced into the polluted environment as a free or immobilized inoculum
should be
– able to degrade specific contamination and survive in a foreign and unfriendly habitat,
– genetically stable and viable, and
– move through the pores in the soil.

– Biostimulation is used in order to modify the physical and

chemical parameters of the soil.
• For this purpose, compounds such as nutrients (e.g. biogas slurry, manure,
spent mushroom compost, rice straw and corncob) or electron acceptors
(phosphorus, nitrogen, oxygen, carbon) are introduced into the soil.

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• Ex-situ Methods
– Constructed wetlands require the competition of microbes
(bioremediation) and plant (phytoremediation).
• Microorganisms degrade or sorb organic substance present in the water
undergoing treatment.
• Plants are used to remove, transfer or stabilize contaminants through
metabolism, accumulation, phytoextraction or immobilization at interface of
roots and soil
– Landfarming is one of the most widely used soil bioremediation
technologies.
• In this technology, excavated contaminated soils are spread out in a thin
layer on the ground surface.
• Aerobic microbial activity within the soil is stimulated through the aeration
and addition of minerals, nutrients and moisture.

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iii. Integrated compost production - microbe
enriched compost
• pH - The pH of the compost pile changes naturally throughout the
composting process.
– Initially, the pH is acid because organic acids are released during
the early stages of decomposition.
– As the temperature rises, the pH increases and becomes slightly
alkaline
• Temperature -Two populations of bacteria in the compost pile that
flourish at different temperature ranges are
• Mesophilic bacteria dominant at temperatures up to 105F and consume the most
readily decomposable carbohydrates and proteins
• Thermophilic bacteria at temperatures from 105 to 160F and initially decompose
proteins and non-cellulose carbohydrates and eventually attack the lipid and
hemicellulose fractions
– High temperatures also are more likely to kill weed seeds and
plant pathogens.

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• Oxygen -The pile is aerated whenever it is turned.
– Poking the pile with a pitchfork occasionally may also help
aerate the pile.
• Moisture - Water is essential for all the organisms in the compost
pile to live and move in.
– Moisture also keeps nutrients in solution so microbes can
assimilate them.
– The moisture content in the compost pile should be as high as
possible, while still allowing air to filter through pore spaces.
– The optimum moisture level is 50 to 60%
– Bacterial activity ceases at 12 to 15%

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iv. Biopesticide/insecticide production
• Land - Land is required for construction of culture and rearing
rooms, processing room, laboratory, office and construction of poly
house etc.
• Building and civil works - Bio-pesticides production involves
rearing of insects.
– The production unit has to be located away from industrial unit to
avoid pollution problems.
– For the proposed installed capacity, an estimated built up area of
about
• 1000 sq mtr is required for group-I (mass production of Trichogramma,
Chrysoperla and Cryptolaemous beetles) &
• for Group-II (production of NPV, Trichoderma and pheromone lures) about
2400 sq.mtr. area is required.
– Among others, the civil structure may be designed to have
separate room for diet preparation, corcera culture, egg
production, host culture etc.

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• Plant and Machinery - No heavy plant and machinery is required
and all the machinery required are locally manufactured
– Racks, trays and other facilities are required for rearing insects.
– Apart from this centrifuge, mixers and some fabricated
equipment’s for insect collection and rearing are required.
– For production of Trichoderma fermenters, laminar flow
apparatus are required.
• Raw material - For rearing of insects special diet is required which
comprises of pulses, vitamins, antibiotics etc.
– For production of Trichoderma molasses-yeast medium, is
required which are available locally.
• Water The water requirement is mainly for feed preparation,
washing, cleaning, drinking etc.
– Water quality should be tested to establish the suitability.
• Power - Power supply is essential for bio-pesticide units.
– Electricity charges under recurring cost are considered
• Manpower - Production of bio-pesticides required skilled
manpower and is labour intensive
– There is need for a number of laborers at each stage of
production.

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v. Fermented products – probiotics and prebiotics
• Development must be done in a culture medium suitable
for biomass production
• The variables involved during the growth of the strain (pH
of the medium, type, and concentration of carbohydrates,
growth phase at harvesting, cell physiological state, etc.)
may significantly affect
– its resistance to biological barriers (gastric acidity and bile salts)
and
– its capacity to interact with immune cells, thus conditioning its
functionality.
• The physiological status of a strain is involved in its
functional properties.

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vi. Stem cell production, stem cell bank

• Physiochemical environment - Physiochemical culture

parameters include pH, dissolved oxygen and carbon dioxide tensions
and temperature.
– Even though almost all expansion cultures are run at pH 7.4, 20
per cent oxygen, 5 per cent carbon dioxide and 37°C, the
proliferation and differentiation of different stem cell cultures have
been found to operate at different optima.
• Nutrients and metabolites - Nutrient and metabolite
concentrations determine cell growth, differentiation and death in a
culture, and therefore should be closely monitored and controlled in
bioprocessing.

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• Growth factors - Growth factors regulate stem cell behaviour
by providing survival, proliferation and differentiation signals to the
cells.
– They have specific functions, both positive and negative in
nature, and can act on either a specific cell lineage or multiple
lineages.
– Interactions between these growth factors and/or with other
process parameters to one another is essential in order to tailor
the culture process for optimal production of a specific cell type
population

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vii. Production of monoclonal/polyclonal antibodies
• Preparation of the antigen - When antibodies are produced, it
is important to consider antigen features, which include the quality and
quantity of the antigen and the antigen preparation.
– The specificity of the immune response obtained depends on the
purity of the antigen applied.
• Selection of the animal species - The most frequently used
animal species for PAb induction in the laboratory setting are the
rabbit, mouse, rat, hamster, guinea pig, goat, sheep, and chicken.
– The selection of an animal species is also influenced by the
species from which the antigen is taken.
– The greater the phylogenetic distance between the animal species
that is the source of the antigen and the species of the animal to
be immunized, the better the immune response that will be
evoked.

25
• Selection and preparation of the adjuvant - When the
antigen to which antibodies are to be evoked is poorly immunogenic,
the immune system requires a stimulus to induce an effective immune
response.
– Adjuvants can be used for this purpose, and can direct an immune
response against a more cellular or humoral response
– More than 100 adjuvants have been described
– Only a few adjuvants are routinely utilized for polyclonal antibody
production and include Freund's complete adjuvant (FCA 1),
Freund's incomplete adjuvant (FIA 1), aluminum salts (e.g.,
Al(OH)3, AlPO4)
• Injection protocol - The choice of injection route is shaped to
some extent by the choice of the animal species and adjuvant, as well
as by the character, quantity, and volume of the antigen.
– Regardless of the mixture to be injected, it is imperative to use the
smallest possible volume that induces a sufficient antibody
response.
– The minimal volume depends on particular characteristics of the
antigen. 26
• Post injection observation - After immunization, animals
should be monitored daily and examined for specific side effects at
least three times per week.
– Examination and palpation of the injection site are essential to
evaluate side effects of the injected mixture.
– In mammals, antibody responses during the experiment can be
monitored by obtaining and evaluating a blood sample for
antibodies in the serum.
– In chickens, because antibodies are excreted in the eggs, titers
can be studied in the egg without an invasive action.
• Collection of the antibodies - Exsanguination must be
performed under general anesthesia and is best carried out by heart
puncture.
– It should result in the death of the animal.
– When there is uncertainty about death, small rodents can be
subjected to cervical dislocation, and larger animals can be
euthanized by an overdose of an appropriate anesthetic agent.

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viii.Single Cell Protein

28
ix Secondary Metabolite Production
• Environmental Factors - The local geoclimate, seasonal
changes, external conditions such as light, temperature, humidity
affect composition of secondary metabolites.
– The synthesis of secondary metabolites, including saponins,
response to environmental factors
• Light - Light can stimulate secondary metabolites like gingerol and
zingiberene production in Z. officinale callus culture
– Moderate light intensity (301-600 lx) induced higher accumulation
of anthocyanins
– UV-B have been seen to increase in flavonoids in barley,78 and in
polyamines in cucumber

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• Nutrient Stress - When plants are stressed, secondary
metabolite production may increase because growth is often
inhibited more than photosynthesis, and the carbon fixed is
predominantly allocated to secondary metabolites.
– The Daucus carota callus subjected to phosphate stress
produced 7.2% dry wt anthocyanin against 5.4% dry weight
(DW) in the control.
– Deficiencies in nitrogen and phosphate lead to the accumulation
of phenyl propanoids and lignification.
– In tomato, the 3-fold increase in anthocyanidins level and the
simultaneous doubling of quercetin-3-O-glucoside occurs under
nutrient stress

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• Temperature - Changing the culture temperature affect
secondary metabolite production.
– Melastoma malabathricum cell cultures incubated at a lower
temperature range (20 ± 2°C) grew better and had higher
anthocyanin production than those grown at 26 ± 2°C and 29 ±
2°C.
– Optimum temperature (25°C) maximizes the anthocyanin yield
as demonstrated in cell cultures of Perilla frutescens
– For strawberry cell culture, maximum anthocyanin content was
obtained at 15°C and it was about 13-fold higher than that
obtained at 35°C.

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 MARKETING STRATEGIES
i. Pollution Monitoring

• The growth of the environmental monitoring market is

driven by factors such as
– growing global population,
– development of policies and initiatives aimed to reduce air, soil,
and water pollution,
– increasing government funding towards pollution prevention and
control,
– growing construction of environmental monitoring stations,
– growing initiative towards the development of environment-
friendly industries, and
– reduction in export tariff on environmental monitoring
technologies in emerging and developed markets.

32
• Strategies include
– Market Penetration: Comprehensive information on
environmental monitoring products offered in the global
environmental monitoring market. It analyzes the environmental
monitoring market by type of product, sensors, sampling method,
and application across key geographic region
– Product Development/Innovation: Detailed insights on upcoming
environmental monitoring technologies, research and
development activities, and new product launches in the global
environmental monitoring market
– Market Development: Comprehensive information about lucrative
emerging markets for various environmental monitoring products
across key geographic region
– Market Diversification: Exhaustive information about new
products, untapped geographies, recent developments, and
investments in the global environmental monitoring market
– Competitive Assessment: In-depth assessment of market shares,
strategies, products, distribution networks, and manufacturing
capabilities in the global environmental monitoring market
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ii. Bioremediation for Industrial pollutants,
Pesticides, Herbicides
• Asian countries showed marked variability between rich and
poorer countries in the perception and approach to
remediation issues.
• These reflected their relative abilities to invest in R & D for
novel techniques such as bioremediation.
• Marked contrasts in levels of importance was accorded to
process-based remediation techniques versus physical
methods like land filling.
• There was a lack of centralized information on remediation
activity, even in some first-world countries, and differences in
levels of funding for the development and provision of
remediation information.
• Thus, economic barriers may limit certain countries’ access to
the growing body of information on degradation of xenobiotics
by micro-organisms.

34
• Marketing strategies depend on
– economics/cost considerations
– relative perceptions or ignorance of the extent of contamination
– country-specific policies and bio-safety legislation and
– stronger focus on an environmental medium such as soil at the
expense of others.

35
 iii. Integrated compost production-microbe
enriched compost
• Transport costs and customers’ ability to pay
• Composting urban organic waste close to the raw waste
source usually leads to the household market: lucrative but
often low-volume.
• However, bulk markets are often located far from
residential areas, requiring transport of either waste or
compost.
• The relative advantage of transporting compost is its
volume and weight: as little as 30 per cent of raw materials
used and is also more hygienic to handle and transport
compost than waste.
• Small towns are an exception; composting plants can be
located in peri-urban areas near the sources of raw
materials and markets.
36
• Strategies include
– Draw a map of your city and its surrounding area;
– Mark the location(s) of compost production unit(s);
– Locate and indicate the organic waste sources (e.g. vegetable
markets and households);
– Locate customers and indicate the distance to customers.
– Use different colours for each market segment;
– Assign demand or quality requirements to each customer; and
– Compare the demands and distribution costs of the different
locations

37
iv. Bio-pesticide/insecticide production
• The development of the bio-pesticide market in the
future is strongly related to research on biological control
agents.
• Several scientists from diverse research institutes have
done some preliminary research in the field, but
complete and systematic reports are scarce.
• Therefore, it is necessary to strengthen the collaboration
of enterprises and research institutes.
• It seems that bio-pesticides cannot as yet completely
replace chemical pesticides, so the agricultural sector
can and should benefit from the co-existence of bio-
pesticides with chemical pesticides.
• In this regard, accelerating practical application of
research results is expected to facilitate large-scale
industrial development. 38
• The strategies are focussed on.
• Supply side: Biopesticide producers, suppliers,
distributors, importers, and exporters
• Demand-side: Farmers, farmer’s organizations, crop
protection products manufacturers, and contract farmers
• Regulatory side: Concerned government authorities,
commercial research & development (R&D) institutions,
and other regulatory bodies
• Associations and industry bodies such as the Food and
Agriculture Organization (FAO), Biopesticide Industry
Alliance (BPIA), International Biocontrol Manufacturers
Association (IBMA), and Association of American Plant
Food Control Officials (AAPFCO)

39
v. Fermented products - Probiotics and Prebiotics
• The probiotic market, especially dairy products such as
yogurts and fermented milks, has experienced rapid
growth in Europe.
• The long-term exploitation of probiotics as health
promoters is dependent on several factors, including
sound, scientifically proven clinical evidence of health-
promoting activity; accurate consumer information;
effective marketing strategies; and, above all, a quality
product that fulfills consumer expectations.
• The strategies include
– Compelling and continuous research.
– Marketing then follows, which raises the question of whether the
marketing aligns with the research.
– Finally, there are regulations, which tend to kick in when the
marketing and research do not line up.
40
vi. Stem cell production, stem cell bank
• Involves the creation of a umbilical cord blood (UCB)
storage bank
• Serial process that includes collecting UCB, extracting
stem cells from UCB, storing these stem cells in liquid
nitrogen at –385°F, and defrosting them if required by
their owner.
• UCB banking service is thus a form of biological
insurance, where the reward is an increased chance of
good health rather than monetary compensation.
• Marketing techniques along with IMC- Integrated
Marketing Communication tools play a crucial role in
creating the awareness, grabbing the attention, arousing
the interest, building the conviction and persuading
people to act favorably
41
• Strategies include
i. Specially Designated Vehicle to propagate the
importance of Cord Blood Stem Cells Preservation:
– A special unique vehicle has to be designated to propagate the
importance of the Stem Cells Preservation and to eradicate the
Myths prevailing among the people.
– This Special vehicle has to contain the following:
• Illustrative display of information about the importance & need for Stem Cells
Preservation,
• A projector system and the screen to exhibit the short film to entice about Stem
Cell Preservation,
• A qualified and trained counselor, informative booklets in vernacular languages

ii. Print Aids:

• Crafting the informative posters (with Cartoon characters),
• Banners (with pictorial descriptions & explanations) &
• Billboards in the places of the public prominence.
• All of the above should be done in English, Hindi and all the
prominent vernacular languages.

42
iii. Audio Visual Aids:
• Short films has to be produced and exhibited at schools, colleges,
universities, offices, and other institutions, where there is a scope of
exhibiting the film. (i.e., subject to availability of projector system and other
infrastructural facilities to exhibit the film).
• Short film should be in English, Hindi and other prominent vernacular
languages to reach the maximum number of people and make people aware
of the importance and need of the stem cell
• blood stem cells preservation etc. with the aim of making people aware, motivated
and prepared to preserve their stem cells.

iv. Advocacy by the Eminent Persons:

• Advocacy about the importance of the voluntary blood donation is very
pivotal as it is a powerful tool to influence the people’s perception about the
stem cell preservation.
• Services of the eminent persons have to be utilized to propagate the need
for voluntary blood donation among the people.

43
v. Awareness Inception Programs:
– In order to incept the awareness about the need and importance
of cord blood stem cells preservation, the following has to be
done:
• Advertisements in Television, Newspapers and other Medias.
• Exhibiting trailers at movie theaters.
• Using posters and stickers to disseminate the idea at the public transport
vehicles like, Buses, Trains etc.
• Posters and banners at the public utility places. Bus stands, railway stations,
airports, hospitals, banks etc.

vi. Trust Building Effort:

• It is observed that more than (80.00 %) of the people are having the opinion
that the quality and maintenance of the stem cell preservation banks not
credible and trustworthy, it is very important to eradicate this myth.
• To achieve this, the ministry of Health, Department of Health and Public
Welfare and other key departments of the government has to disseminate
the information, that the stem cell preservation banks are under the strict
scrutiny of the government and their activities are monitored at most
vigilance.

44
vii. Production of Monoclonal / Polyclonal
Antibodies
• Global antibodies market strategies involve
– continuous launch of innovative products,
– high adoption rate of therapeutic antibodies in cost-sensitive
markets
– quick approval by regulatory authorities for breakthrough
therapies
– rise in chronic illnesses owing to population shift and increasing
longevity
– availability of inexpensive biosimilar antibody therapeutics and
– adoption of diagnostic antibodies.
• Companies are adopting various strategies, such as
– expansion of product indications to sustain in the dynamically
growing antibodies market.

45
viii. Single Cell Protein

• The global single cell protein market is expected to

witness considerable growth in the near future
attributable to growing demand for food supplements as
a result of increasing number of population with
malnutrition.
• The strategies include
• Market Segmentation - Done by species, by feedstock, by
application, and by region.
– By species the global single cell protein market is
segmented into, Yeast, Fungi, Bacteria.
– By feedstock, the global single cell protein market is
segmented into organic and inorganic.

46
 – By application, the global single protein market is segmented in
to, food and beverages, animal feed and pet food, dietary
supplements and others segments.
• The food and beverages segment is further sub segmented into,
fortified food and fortified beverages.
– On the basis of a region the global single cell protein market is
segmented into, North America, Latin America, Western Europe,
Eastern Europe, Asia Pacific excluding Japan, Japan and the
Middle East and Africa.
– North America and Western Europe are expected to account for
significant revenue shares in the global single cell protein market
• Market Drivers and Trends- Increasing number of malnutrition
cases across the globe is driving the market for single cell protein.
– Data from UNICEF states that globally half of the deaths in
children under age five are due to malnutrition or under nutrition.
– For example, between 2010 and 2016, 35.8% of the global
children population in South Asia is under nutrition followed by
34.5% in eastern and southern Africa as per data revealed by
UNICEF.
47
• Market Key Players - Some of the key players offering single
cell protein include
– NOW Food Health LLC.,
– Willows Ingredients,
– Devenish Nutrition Limited,
– BIOMIN Holding GmbH,
– PRO SOLO SPA,
– Aumgene Biosciences,
– BIO-CAT,
– Novozymes,
– Alltech, Inc.,
– Nutreco N.V.

48
ix Secondary Metabolites

• The growing demands in today’s market place for

renewable natural products lead scientist to consider in
vitro plants material cultures as an alternative method for
production of secondary phytochemicals.
• Bulk of the market products such as secondary
metabolites from higher plants are collected in the wild,
providing an opportunity for developing production
strategies that are environmentally sustainable and
economically viable
• Industrial production of secondary metabolites is now
only commercially viable when the market value exceeds
U.S. one-thousand to several thousand dollars per
kilogram of product.

49
 SCHEMES
i. Pollution Monitoring
• Pollution monitoring schemes are usually controlled by
Ministries and research councils.
• Unfortunately these are often ill designed
• For eg. Nature Conservancy measures pollutants in
plants and animals yet the value of results in assessing
pollution control policy was not major consideration when
programme was drawn up
• Most of the actual measurements are carried out by
– Local government
– Individual Universities
– Companies and
– Other organizations
• Hence decentralized schemes for pollution monitoring
should be integrated to have pollution control
50
ii. Bioremediation for Industrial pollutants,
Pesticides, Herbicides
• Biostimulation (BS) by supplementing with slow-release
nitrogen and phosphorus fertilizers
• Bioaugmentation (BA) by inoculating with Cycloclasticus
sp. E2, an aromatics-degrading bacterium identified to
play an important role for aromatic-hydrocarbon
degradation in marine environments and
• Combination (CB) of BS and BA
• Bioaugmentation with ecologically competent pollutant-
degrading bacteria is an ecologically promising
bioremediation scheme.

51
iii. Integrated compost production-microbe
enriched compost
• Key aspect is marketing and promoting city compost.
• The Department of Fertilizers involved existing fertilizer
companies to market compost through their dealer networks.
• Compost is also co-marketed by tagging of cities with fertilizer
marketing companies.
• Only 14 states had been tagged with fertilizer companies for
the implementation of the scheme, but even so, most fertilizer
companies had no marketing presence in these states.
• Entrepreneurs are offered incentives to set up compost
manufacturing facility.
• For example, under National Program for organic farming,
manufacturers of compost from vegetable waste are offered a
subsidy of 33% of the cost of project.

52
• Government is encouraging the production of organic
manure from bio-waste under the Capital Investment
Subsidy Scheme (CISS) of “National Mission for
Sustainable Agriculture (NMSA)” programme.
• Under this scheme, financial assistance is provided
through NABARD (National Bank for Agriculture and
Rural Development) as capital investment for setting up
of mechanized Fruit/Vegetable market waste/Agro waste
compost production unit of 3000 TPA production
capacity.
– 100% upto a maximum limit of Rs.190 lakh to State Govt./Govt.
Agencies and
– 33% upto a maximum limit of Rs.63.00 lakh per unit to
individuals/private agencies

53
• Government is encouraging the use of organic/bio
fertilizer through various schemes/programmes
– National Mission for sustainable Agriculture (NMSA)
– Paramparagat Krishi Vikas Yojana (PKVY),
– Rashtriya Krishi Vikas Yojana (RKVY),
– Mission for Integrated Development of Horticulture (MIDH),
– National Mission on Oilseeds & Oil Palm (NMOOP),
– National Biogas and Manure Management Programme
(NBMMP),
– Network Project on Organic Farming of Indian Council of
Agricultural Research (ICAR) and
– National Programme on Organic Production (NPOP) of
Agricultural & Processed Food Products Export Development
Authority (APEDA).

54
iv. Biopesticide/insecticide production
• 100% Assistance to State Govt/Govt. Agencies upto a
maximum limit of Rs.160.00 lakh /unit and
• 25% of cost limited to Rs.40 lakh/unit for individuals/
private agencies through NABARD as capital investment
of 200 TPA production capacity

55
v. Fermented products-probiotics and prebiotics
• Along with the price revision, the contents of Yakult are
enhanced so this does not merely constitute a simple
price increase.
• First substantive price revision for Yakult in 22 years and
in November 2013 manufacturer’s list price of Yakult was
raised from ¥35 to ¥40 per bottle.
• The fermented milk drink Yakult and other popular dairy
products are now consumed at a rate of over 30 million
bottles per day around the world.
• Schemes include quantitative goals (Fiscal year 2020)
– Number of bottles of dairy products sold worldwide: 36.5 million
bottles per day
– Consolidated sales: 500.0 billion yen
– Consolidated operating income: 50.0 billion yen

56
 vi. Stem cell production, stem cell bank
• From just a few Institutes in India two years back, today
over 30 institutions are involved in stem cell research with
the Indian government investing around $8 million dollars
in just the last two years.
• The Tamil Nadu Government has given a grant of Rs 9
crore to the Public Cord Blood Bank established by the
Jeevan Blood Bank and Research Centre in Chennai
to process and store about 3,000 cord blood donations
from Tamil Nadu.
• There are almost 180 prominent companies working on
stem cell research in the world, majority of which are
based in the US, followed by the EU, Israel, Thailand,
Canada, and Australia.
• India and China are poised to play a key role in the
scientific, clinical and commercial development of stem
cell research. 57
vii. Production of monoclonal/polyclonal
antibodies
• The largest cGMP-compliant plant-based production
process for mAbs has been established by the
Fraunhofer IME (Aachen, Germany) for the manufacture
of 2G12.
• This facility can process 250 kg of tobacco leaf biomass
per batch, corresponding to approximately 900 L of bulk
extract
• The potential integration of simulated moving bed
chromatography and continuous processing operations
to reduce the size and footprint of the equipment,
increasing the space–time yield for the single-use
continuous manufacturing of mAbs
• The reduction in capital costs of 10–30%, which is
particularly striking 58
viii.Single Cell Protein
ix. Secondary Metabolite Production

• The world wide, large-scale research and development

of SCP processes has involved work in the fields of
microbiology, biochemistry, genetics, chemical and
process engineering, food technology, agriculture,
animal nutrition, ecology, toxicology, medicine and
veterinary science and economics.
• SCP processes are used in related technologies e.g. in
waste water treatment, production of alcohol, enzyme
technology and nutritional science
• The future of SCP will be heavily dependent on reducing
production costs and improving quality.

59
• Large scale production and commercialization of
valuable Secondary Metabolites from callus derived cells
of vascular cambial explants of selected woody plant
species have been funded by PACE (Promoting
Academic Research Conversion to Enterprise) scheme

60
 CHALLENGES AND SCOPE
i. Pollution monitoring

• To monitor the Industry emissions in real time, the need

is to have a Highly Scalable Platform that can expand
the needs to all the polluting industries across the
country.
• With the concept of Smart City taking centre stage in the
country, the amount of sensors that will be deployed to
monitor and control environmental parameters is
estimated to be in millions.
• The need of the hour is to have a plug and play platform
where new sensors could automatically join the grid and
contribute seamless for effective pollution control.

61
• The current data gathered for online monitoring is used
for only complying to regulatory standards, however, the
strength lies in the leveraging this data and make the
use of the new advances in Data Sciences and Big Data
Analytics to make real time decisions and controls to
effectively control pollution.
• Data is in plenty but quality of data is another key
challenge when dealing with the sensor or analyzer data.
• Automated pattern mining and establishing automated
quality validation on the data can now be easily
implemented using machine learning algorithms.
• Another key challenge is with respect to pollution
prediction.

62
• Scope
• Data about current pollution or historical pollution is
available but scope lies to accurately predict or forecast
the pollution levels based on the previous observations.
• Once we are able to successfully predict and forecast,
there is a good possibility of influencing these levels
leveraging the citizen and industry participation.
• Establishing the correlation of traffic impact on Pollution,
there could be intelligent traffic routing during peak hours
to reduce environment pollution at various strategic
points by reducing the traffic jams.

63
ii. Bioremediation for Industrial pollutants, Pesticides,
herbicides
• Deficit of knowledge: A deficit of knowledge on the
different fundamental branches of sciences that are
involved in the process of bioremediation hinders the
progress of bioremediation.
– Examples of the different disciplines of sciences are structural
and molecular biology, microbiology, genomic, geo – chemistry,
along with hydrology and transport processes.
• Apart from that, the pathways of biodegradation and
biotransformation remains unclear.
• There is still much to learn and gain from research as
contaminated sites are complex systems – each
composed of different types of contaminants, diverse
organisms, and similar environment.

64
• Lack of integrated research: Bioremediation is a
multidisciplinary field and researchers from various fields
need to integrate their knowledge.
– The success and efficiency of bioremediation requires the
involvement of microbiologists, biochemists, engineers,
geologists, and soil scientists among others.
• Lack of revenue: There is no doubt that government and
private sectors are investing on biotechnology
companies but not much of he revenue goes to the
improvement of bioremediation.
• Inadequate tools and infrastructure: Accuracy in
experimental procedures is crucial to ensure successful
treatment when introduced to contaminated site.
– However, there is inadequacy of infrastructure to analyze the
conditions of contaminated sites as well as monitor the process
of degradation that has been introduced to the site.
65
• Scope
– Environmentally pleasing technology
– No pollution
– No toxic residues

66
iii. Integrated compost production- microbe enriched
compost
• Policy framework about composting urban organic solid
wastes: Political will must exist to formulate and implement policy
promoting composting in line to enhance participation in terms of
household waste separation
– Hence, not in my backyard can present a problem both in
developed and developing countries when selecting waste
management sites.
– Municipal authority should have management structure and
framework of legislation for effective composting of organic solid
wastes.
• Budget or financial constraint: After taking into consideration the
expense of compost (US $5.80 per ton of compost, or US $3.08 per
ton of waste processed) and recyclable sales, the municipality is
required to subsidize the operation at a cost of US $10.76/ton of
waste processed.
– However, financial issues are the main problems for most of the
developing countries
67
• Lack of department of urban agriculture (institutional
problem): There is no urban agriculture department in the town to
encourage composting urban agricultural activities to increase the
fertility and water holding capacity of the soil.
– urban farmers engaged in grain production activities using
organic waste dumped in their field and backyard decomposed
after a period of time as a soil conditioner.
– Hence, the establishment of the department of urban agriculture
was depending on the policy framework.
• Lack of knowledge about compost preparation: most of
the urban farmers were not aware of right way of compost
preparation.
– Due to this, most of the respondents engaged in agriculture uses chemical
fertilizer to improve soil fertility and made an additional expenditure

68
• Lack of knowledge about the advantage of compost over
the chemical fertilizer: compost provides a stable organic matter
that improves the physical, chemical, and biological properties of
soils, thereby enhancing soil quality and crop production;
– When correctly applied, compost has valuable effects on soil
properties, thus creating suitable conditions for root development
and consequently promoting higher yield and higher quality of
crops

• Scope
– Improves soil structure and soil aggregate stability, resulting in
better drainage, aeration, air/gas exchange, erosion resistance,
tilth (workablity), and the soil’s ability to recover from
compaction.
– Increases moisture retention
– Provides a slow-release source of nutrients and increases
availability of minerals.
– Increases the population and diversity of microbes in soil that
continually make nutrients available to plants.
– Plays key role in soil fertility management in organic systems
69
iv. Biopesticide/insecticide production.
• Political - The political factors hindering biopesticide development,
adoption and diffusion are centred on government policies
– The poor enforcement of pesticide regulation is hampering the
development, adoption, and diffusion of biopesticides
– Furthermore, this has hampered investments in knowledge
development, marketability, and accessibility to biopesticides
• Technological - The technological factors hindering biopesticide
development are centred on the lack of solid research and development
infrastructure in the country.
– The academia and industry lack the requisite scientific knowledge
and technological skills to research, develop, and commercialize
biopesticide products.
• Socioeconomic - The socioeconomic factors hampering biopesticide
development and widespread utilization are centred on the cost and
social acceptability.
– low government promotion and patronage has severely impacted
on the availability and acceptability of biopesticides
70
• Scope
– The interest in organic farming and pesticide residue free
agricultural produce would certainly warrant increased adoption
of biopesticides by the farmers.
– As environmental safety is a global concern, manufacturers,
government agencies, policy makers and the farmers have to
switch-over to biopesticides for pest management requirements.
– It is also believed that biological pesticides may be less
vulnerable to genetic variations in plant populations that cause
problems related to pesticide resistance.
– If deployed appropriately, biopesticides have potential to bring
sustainability to global agriculture for food and feed security.

71
v. Fermented products-probiotic and prebiotics.

• The search for economical and abundant raw materials

for prebiotic production and the low-cost production of
probiotics is challenge
• Retention of viability of the probiotic bacteria for
application of probiotic cultures in functional foods
• Many active cultures die during manufacturing, storage
or transport of the finished product and also during the
passage to the intestine
• A serious problem of shelf instability had been
encountered with dried cultures.
• Refrigerated products also have short lives due to
negative effects of low temperature and formation of
crystals on bacterial cells.

72
• The numbers of viable bacteria continually decrease with
time during refrigerated storage
• Manufacturers of probiotics are facing the challenge that
they should produce probiotic cultures that can survive
for long periods, and are resistant to acidity in the upper
intestinal tract so that they can reach the colon in high
numbers to colonize the epithelium.
• The preservation of these probiotic microorganisms
presents a challenge because they are affected by
exposure to temperature, oxygen and light

73
• Scope
– The traditionally fermented fruits and vegetables not only serve
as food supplements but also attribute towards health benefits.
– Lactic acid fermentation is expected to play an important role in
preserving fresh vegetables, fruits, and other food items for
feeding humanity in developing countries
– Lactic acid fermentation increases shelf life of fruits and
vegetables and also enhances several beneficial properties,
including nutritive value and flavours, and reduces toxicity.
– Fermented fruits and vegetables can be used as a potential
source of probiotics as they harbour several lactic acid bacteria
such as Lactobacillus plantarum, L. pentosus, L. brevis, L.
acidophilus, L. fermentum, Leuconostoc fallax, and L.
mesenteroides.

74
vi. Stem cell production, stem cell bank
• To develop large collections of induced Pluripotent Stem
cell lines, while maintaining the cell and data quality
necessary for high-level research.
• New cell culture techniques and approaches are needed,
as well as methods for rapid characterisation of the cells
to provide the necessary data on the exact nature of the
cells that are being grown and banked.
• Stem cell banks will have a serious responsibility for
preventing the repeat of past problems, where cells lines
have been mixed up or contaminated with microbes,
leading to publication of misleading research findings.
• In addition, stem cell banks will need to play a role in
ensuring that any cells used in research have been
sourced ethically - an important issue in sustaining public
support for stem cell research in general.

75
• Scope
– Pharmaceuticals: diabetes, cardiac anomalies,
neurodegeneration, and infertility
– Neutraceuticals: produce heterologous nutrients
– Understanding early embryonic development
– Model for in vitro drug and immunity screening
– Tissue remodeling and engineering
– Enormous potential of renewing body tissues
– Conservation

76
vii. Production of monoclonal / polyclonal antibodies
• Low yield: In vitro production cultures often yield less
antibody/ml than the in vivo ascitic fluid method.
• Cell death: Maintaining optimal cell growth and viability
is essential for high antibody yields and an optimized
process.
• Contamination: In vitro production cultures are subject
to risks of contamination associated with long-term cell
cultures.
• Time-consuming: In vitro production cultures often have
to be maintained over a long period of time in order to
yield desired quantities of antibodies.
• Scalability: Translating research scale antibody
production to large-scale commercial production can be
difficult. Therefore, one must consider scalability of
process when optimizing a production process. 77
• Cost: Components of in vitro production systems (such
as equipment, consumables, and media formulations)
can be very expensive.

• Scope
– Monoclonal antibodies are of immense importance in the field of
oncology especially in the typing and classification of
malignancies and identification of malignant cells, tumour
imaging, chemotherapy and identification and characterization of
significant tumour antigens for serodiagnosis and perhaps
tumour therapy.
– Choice of target antigens for monoclonal antibodies and of labels
to the monoclonal antibodies for the various applications has
been discussed.

78
viii. Single Cell Protein

• Substrate costs: The substrate costs are the largest single cost
factor. Simplifying the manufacture and purification of raw material
can save costs.
– Factors involved in the raw materials costs are site, raw material
production, process capacity of the plant and substrate yield.
• Utilities: The energy for compressing air, cooling, sterilizing and
drying forms the next most important cost factor.
– Sites with cheaply available thermal, electrical, fossil or process
derived energy are to be preferred.
• Product-specific variables: The process costs arising are
covered only by the product produced.
– The absolute value of the product is governed by the amount of
product referred to the costs involved and by the quality of the
product.
– The upgrading of the product may consist of purification and
separation of the microbial biomass.

79
• Capital load: The capital dependent costs are determined, by the
cost of the apparatus for the process, the capacity of a plant and the
capacity conditions.
– The main variable here is the size of the plant. Small plants can
be profitable only if they include simplifications of processes and
material to a considerable degree.
– The greater expenditure on apparatus in processes with cheap,
simple and unpurified raw materials usually does not pay in
comparison with more expensive pure substrates with simpler
technology.
– High productivities in fermentation are compensated by the
greater expenditure on energy to achieve these productivities, so
that optimum can be determined.

80
• Scope
– Microalgae are currently used mainly in the form of supplements,
available in tablet, capsule or liquid form, but they are
increasingly also processed as ingredients which can be
included in pastas, baked goods, snacks, and so on
– Algae are primarily used in aquaculture as a source of omega
fatty acids and carotenoid pigments, but their protein also
contributes to animal nutrition
– SCP derived from fungi is expected to provide vitamins primarily
from the B-complex group (thiamine, riboflavin, biotin, niacin,
pantothenic acid, pyridoxine, choline, streptogenin, glutathione,
folic acid, and p-amino benzoic acid)
– Bacterial SCP generally contains 50–80% protein on a dry
weight basis and the essential amino acid content is expected to
be comparable to or higher than the FAO recommendations.
– Methionine content up to 3.0%, which is higher than that
generally obtained in algal or fungal SCP

81
ix. Secondary Metabolite Production

• Increasing the productivity of cultured plant cells and the

necessary metabolite through standardization of culture
environment
• Manipulation of plant cell cultures to improve productivity
of target compounds employing elicitors, abiotic stresses
and other approaches, regardless of their mechanisms
• Studying signal transduction pathways underlying
various effective strategies leading to the biosynthesis of
target secondary metabolites

• One Major limitation in the production of secondary

metabolites by plant cell culture technology is the low
yield of secondary metabolites.
82
• Scope
– Secondary metabolites are economically important as drugs,
flavor and fragrances, dye and pigments, pesticides, and food
additives.
– Secondary metabolites find wide scope and applications in the
field of agriculture and animal health as follows:
• Kasugamycin and polyoxins are used as biopesticides;
• Bacillus thuringiensis crystals, nikkomycin, and spinosyns are used as
bioinsecticides;
• Bialaphos find application as bioherbicides;
• Ivermectin and doramectin are used as antihelmintics and endectocides
against worms, lice, ticks, and mites;
• Ruminant growth promoters are in the form of coccidiostats;
• Plant hormones like gibberellins are used as growth regulators

83
 CONTACT RESEARCH IN MICROBIAL
GENOMICS
• The study of genomes as an entity as opposed to
individual genetic components is referred to as
genomics.
• Microbial genomes encompass all chromosomal and
extra chromosomal genetic material.
• Microbial genomes are widely variable and reflect the
enormous diversity of bacteria, archaea and lower
eukaryotes.
• Bacterial genomes usually consist of a single circular
chromosome

84
• Species with more than one chromosome (eg.
Deinococcus radiodurans), linear chromosomes (eg.
some Bacillis subtilis strains) and combinations of linear
and circular chromosomes (eg. Agrobacterium
tumefaciens) also exist.
• The research of microbial genomes helps
– To better understand the broader biology of bacteria, and how
their genetic composition contributes to their tangible
characteristics.
– To infer the evolution of bacteria.
– To determine the origins of bacteria and map the transfer of
genes such as those conferring antibiotic resistance.

85
• Microbial Genomics can be studied using the following
techniques
a) Pulsed Field Gel Electrophoresis (PFGE)
• The principle underlying PFGE is very similar to normal
gel electrophoresis.
• PFGE allows to resolve far large, 'genome scale' pieces
of DNA (greater than 20 kilobases in size).
• PFGE is still an important technique used to estimate the
size of microbial genomes and in epidemiology studies.

b) DNA Sequencing
• DNA sequencing has been an enormous advancement
in the field of microbial genomics to amass vast amounts
of genetic data from our organisms of choice.

86
• Now there are methods for sequencing entire bacterial
genomes with relative ease.
• Whole genome sequencing produces immense amounts
of data from which a catalogue of important information
can be derived.

c) Bioinformatics
• The huge amounts of genomic data produced through
DNA sequencing can often be very confusing and
difficult to analyze.
• Therefore bioinformatics is often required to derive
useful information from this data like
– Map the positions of genes,
– Elicit their functions and
– Infer evolutionary relationships
87
d) Microarray-based genomic technology
• Microarrays (or microchips) are a recently developed
genomic technology and are listed as one of 10
breakthrough technologies in 1998, along with
genomics
• Microarray-based genomic technologies represents a
powerful new tool that allows researchers to view the
living cell under various physiological states from a
comprehensive and dynamic molecular perspective and
have revolutionized the genetic analysis of biological
systems.

88
• The widespread, routine use of such genomic
technologies will shed light on a wide range of important
research areas, such as
– How cells grow, differentiate, and evolve;
– The medical challenges of pathogenesis, antibiotic resistance,
and cancer;
– Agricultural issues of seed breeding and pesticide resistance;
– The biotechnological challenges of drug discovery; and
– The remediation of environmental contamination.

e) Proteomics.
• Recent progress in whole-genome sequencing has
provided information on the composition of many
proteins from a variety of organisms

89
• To understand how a proteome (i.e., a complete set) is
produced and the proteins interact with each other, high-
throughput proteomic approaches such as mass
spectrometry (MS), phage display, and protein arrays
must be employed.

f) Cellular modeling
• Predicting metabolic activities and cellular behavior is an
ultimate goal of genome-based biology.
• However, modeling global cellular behavior is a major
challenge due to the complexity of metabolic pathways
and lack of understanding of the dynamic behavior and
regulatory mechanisms.
• The most thorough way to analyze metabolic networks is
through a dynamic model of cellular metabolism.
90
• Microbial genomics-related research programs include
– The Microbial Genome Program
(www.ornl.gov/microbialgenomes/index.html)
– The Microbial Cell Project (www.microbialcellproject.org)
– The Natural and Accelerated Bioremediation Research Program
(www.lbl.gov/NABIR)
– The Experimental and Computational Structural Biology Program
(www.science.doe.gov/ober/msd_struct_bio.html)
– An ambitious new program, Genomes To Life (GTL;
www.DOEGenomesToLife.org)
– The Institute for Genomic Research in Rockville, Md., has the
Comprehensive Microbial Resource (CMR)
(http://www.tigr.org/tigr-scripts/CMR2/CMRHomePage.spl).

91