Applied Biology L1

Applied Biology
Dr Srinivasan M
Dr M Srinivasan 1
Syllabus
• Bioprocess technology for the production of cell biomass and
primary/secondary metabolites
• Regeneration of plants and Totipotency, Hairy root cultures and their
cultivation, Transgenic plants in crop protection and their application in
agriculture – Herbicide tolerant plants, Disease resistant plants, Insect
tolerant plants Plant products of industrial importance,
• Cloning in animals, Genetic engineering, transgenic animals, Gene
therapy, Genetically manipulated organisms and products
• GM Foods and current global situation, Pharmaceuticals, Drug Delivery
Books
1. Thieman William J, Introduction to Biotechnology, 2nd edition (2009),
Pearson Education, San Francisco, ISBN-9780321589033
2. Sobti RC, Essentials of Biotechnology, 2009, Ane Books, New Delhi, ISBN-
9788180521607
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Bioprocess Technology
• Use of living cell or their components to obtain
desired product
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Nutrients
Metabolite
Substrate/
Biomass Cell Enzymes Biomass
Oxygen or
CO2/CH4
Light
Input Bioconversion Output

Cheaper, Easily Available Bioreactor Product
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Dr M Srinivasan 6
• Dried cells of micro organisms
Bioprocess • Dietary supplement – animals and humans
Technology – • Pruteen – Methylophilus methylotrophs, cultured on
Single Cell Protein methanol
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Bioprocess Technology – Single
Cell Protein
• Production of SCP Fermentation
• Requires micro-organisms that Substrate along with
other Nutrients
serve as the protein source
• Substrate that is biomass on
which they grow
Filtration Drying
• Fermentation or growing of
microbes
• Harvesting
• Down stream processing SCP
• Processing and Packaging
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Cell Protein
• Micro-organisms
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Cell Protein
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Cell Protein
• Properties
• High growth rate
• High protein content in the dry mass
• Broad spectrum of substrate or biomass
• Growth independent of season changes
• Should not accumulate toxins and heavy metals
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Cell Protein
Lignin
• Substrate Cellulose
• Cheap
• Easy avaialblity
• O2 requirement
• Heat production
Methanol
• Down stream Ethanol
processing cost Gas oil
Dairy waste
Molasses
Cow dung
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Cell Protein
• Fermentation
• Trenches , open pit
• Bioreactors
• Fed batch culture
Lab Scale Bioreactor
Algal Cultivation Dr M Srinivasan Industrial Scale Bioreactor 13

Cell Protein
• Down Stream Processing
Washing
Centrifuge
Fermented To clear left over
To settle down cells
Media and remove liquids
biomass or media or
substrate
In case of Algae,
protein release Centrifuge
Freeze drying
induced by breaking
cell wall
Mechanical, Chemical
Physical
Packing
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Cell Protein – Disadvantages
• High content of nucleic acids leading to elevated levels of
uric acid, kidney stone and gout
• Possibility for the presence of secondary toxic metabolites
• Poor digestibility
• Hypersensitivity
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Bioprocess Technology – Type of
Fermentation
• Batch Fermentation
• Continuous Fermentation
• Fed Batch Fermentation
• Anaerobic Fermentation
• Aerobic Fermentation
• Surface Fermentations
• Submerged Fermentations
• Solid State Fermentation
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Fermentation
• Closed system
• Prepare medium,
sterilize it and inoculate
into the bioreactor
• Allow the cells to grow
and produce the
product
• Once the product
formation reaches
maximum harvest the
broth
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Fermentation
• Continuous
Fermentation
• Fresh nutrient medium
is added continuously
or intermittently
• Equivalent amount of
used medium with
microorganisms is
withdrawn continuously
or intermittently
• Recovery of cells or
fermentation products
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Fermentation
• Fed Batch Fermentation
• Substrate is added periodically in
instalments
• Secondary metabolites are subjected to
catabolite repression by high
concentration of either glucose, or other
carbohydrate or nitrogen compounds
present in the medium
• The critical elements of the nutrient
medium are added in low amount in the
beginning of the fermentation
• These substrates continue to be added in
small doses during the production phase
• This method is generally employed for
the production of substances such as
penicillin
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Fermentation
• Surface Fermentations
• The substrate may be solid or liquid
• The organism grows on the substrate and draws the
nutrients from the substrate.
• These types of fermentations are desirable where
the products are based on sporulation
• But it has several disadvantages such as it exposes
the organism to unequal conditions, both oxygen
and nutrients.
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Fermentation
• Submerged Fermentations
• Solid State Fermentation
• Solid substrate
• Absence or near the absence of free water
• Inert supports can be used
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Bioprocess Technology –
Metabolites
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Metabolites
Primary Metabolite Secondary Metabolite
• Performs physiological • Derivative of primary

function metabolite
• Growth and development • Secondary functions like
• Produced during growth defence, competition and
phase in large quantity attraction
• Basic requirement • Produced during stationary
• Nucleotides, Sugars, Amino phase in low quantity
acids, Vitamins etc • Not a basic requirement
• Pigment, Drugs, Antibiotics,
Essential oils etc
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Metabolites
catechins, theaflavins, tannins
Caffeine, Flavonoids, tannins
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Bioprocess Technology – Penicillin
• Most penicillin's form filamentous broths
• This means they can be difficult to mix due to their high viscosity.
• The increasing viscosity of the broth can hinder oxygen transfer.
• Aerobic
• Optima pH 6.5
• Doubling time 6H
Penicillium notatum
Penicillium chrysogenum
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• Reactor type: Airlift Reactor (bubble column)
• pH 6.5, Temperature 20-24 °C
• Oxygen , Nitrogen (Yeast Extract)
• Carbon source - corn steep liquor 8.5 % ,Glucose
1%, Lactose,
• Substrate - phenylacetic acid
• Calcium Carbonate 1%, Sodium hydrogen
phosphate 0.4%,
• Antifoaming agent: vegetable oil
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• Secondary metabolites are only produced in times
of stress when resources are low and the organism
must produce these compounds to kill off its
competitors to allow it to survive
• It is these conditions that we wish to duplicate in
order to achieve the maximum amount of product
from our fermentation.
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• Primary metabolism - Media for this stage will be
focused on achieving maximum growth and biomass
production.
• Once the desired biomass has been achieved, starve
(Limiting the amount of C and N available to the
culture) the culture and induce the kind of stress
conditions that trigger the production of the antibiotic.
• Use the fed-batch method to feed the culture. As stated
above, this allows us to add the substrate to the reactor
in small increments and to even change the substrate if
we so desire
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After about 40 hours, penicillin begins to be secreted by the

fungus. After about 7 days, growth is completed, the pH rises
to 8.0 or above and penicillin production ceases
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• The resulting penicillin (called Penicillin G) can be
chemically and enzymatically modified to make a
variety of penicillins with slightly different
properties.
• These can be semi-synthetic penicillins, such as;
Penicillin V, Penicillin O, ampicillin and amoxycillin.
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Bioprocess Technology – Ethanol
• Ethanol – industrial and pharmaceutical Solvent &
Fuel, alcohol thermometer, as a disinfectant,
preparation of essence and flavors
• Bacteria (Clostridium and Streptococus sps)
and Yeast (Saccharomyces) can produce ethanol
• High growth and fermentation rate
• High product yield, Osmotolerance, LowpH, High
Temp, with stand chemical and physical stress,
Ethanol and Glucose tolerance
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• Raw Material:
• Sugar, Starch (Grains, Potato) and Cellulose
• Molasses – Beet (15% fermentable sugar, 40-50 l per
ton) or cane (50% fermentable sugar, 200-300 l ethanol
per ton)
• Grains (Starch) – Malting, Milling, Mash and Conversion
with enzymes or acid or alkali treatment (maltose)
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• Separation for a mixture of liquids
• It relies on differences in B.P of component liquids
to be separated
• The mixture to be separated is added to the
distilled pot & is heated
• Low B.P will vaporize first. this vapors passes into
distilling head & then into condenser.
• With in the condenser the vapor is cooled & it
liquefies.
• The resulting liquid is then collected in receiving
flask
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Applied Biology L1

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Applied Biology L1

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Applied Biology

Input Bioconversion Output

Lab Scale Bioreactor

Algal Cultivation Dr M Srinivasan Industrial Scale Bioreactor 13

• Performs physiological • Derivative of primary

catechins, theaflavins, tannins

Caffeine, Flavonoids, tannins

After about 40 hours, penicillin begins to be secreted by the

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