BIOINDUSTRY APPLICATIONS

© Kalasalingam academy of research and education
UNIT-4:
BIOLOGY AND ITS
INDUSTRIAL
APPLICATIONS
BIOLOGY FOR ENGINEERS-BIT21R101
Prepared by
Dr. G. Nadana Raja Vadivu-Week 10
Dr. J. Kanimozhi- Week 11
Dr. B.Vanavil- Week 12

Course Outline
CO 1
CO 2
CO 3.
CO 4. Understand various industrial
applications and products of biology
CO 5.
Course description
BIOLOGY FOR ENGINEERS course deals with
fundamentals of cells, cell cycle and
description of the structure and function of
different parts of a cell. This course helps to
identify the different biomolecules. It also has
components dealing with molecular structures
like DNA, its discovery and the processes
involved in the central dogma of Molecular
biology. The course also provides a detailed
description of Microbes as infectious agents
and types of immunity and comprises the
applications of biology in various industries.
COURSE NAME: BIOLOGY FOR ENGINEERS-BIT21R101

Unit 4
Outcomes:
Understand various industrial applications and
products of biology
Syllabus
Unit IV: Biology and its Industrial
Applications
Week 10
Probiotics
Enzymes
Biofertilizers
Biomaterials
Week 11
Bioenergy
Waste water treatment
Role of Genetic Engineering: Insulin
Antibiotics
Week 12
Vaccines
Monoclonal antibodies
Stem cell Technology
Self healing concrete

BIOLOGY FOR ENGINEERS-
BIT21R101 UNIT-4- WEEK 10

Course Progress-Week 10
Lesson 2. - Biofertilizers
Lesson 1. Probiotics and Enzymes
Lesson 3. Biomaterials and Bioenergy

Probiotics
Beneficial Bacteria
• Probiotics
• Nitrogen Fixation
https://images.app.goo.gl/FzqyPWbLzsVhGVnSA

Probiotics
Beneficial microorganisms
 Helpful bacteria and fungi that are either added or naturally
occur in foods.
 Create unique flavors
 textures or improve digest foods or fight disease

Probiotics
Bacteria
• The most important bacteria used in food production are the
Lactobacillaceae family
• Lactic acid from carbohydrates, resulting in changes in certain
foods
• Example: milk to yogurt.

Probiotics
BENEFICIAL BACTERIA - PROBIOTIC BACTERIA
 Many proposed, however: a live microbial feed supplement which beneficially
affects the host animal by improving its intestinal balance
 Historically: terrestrial animals, genus Lactobacillus
Definition
 bacteria in aquatic medium influence composition of gut microbiota and vice versa
 Immediate ambient environment has much greater influence on microbiota than
with terrestrials
 In aquatic environments, hosts and microorganisms share the ecosystem
 Terrestrials: the gut represents a moist habitat in a water-limited world

Probiotics
Yeast
• The most beneficial yeasts for food production are from the genus Saccharomyces
• Yeasts produce desirable chemical reactions
• Example: leavening of bread and production of alcohol
Molds
• Molds from the genus Penicillium are associated with the ripening and flavor of
cheeses.

Probiotics
https://images.app.goo.gl/k5amHPJBHBdaZMJi9

Probiotics
https://images.app.goo.gl/oHrZuku2bnfYx1PT7

Probiotics
https://images.app.goo.gl/mS5joSMrbA7gP4Xu5

Probiotics
Characteristics of Effective Probiotics
 Able to survive the passage through the digestive system
 Able to attach to the intestinal epithelia and colonise
 Able to maintain good viability
 Able to utilise the nutrients and substrates in a normal diet
 non pathogenic and non toxic
 Capable of exerting a beneficial effect on the host
 Stability of desired characteristics during processing, storage and transportation
 Anti-inflammatory, anti-mutagenic and immunostimulatory

Probiotics

Probiotics
Fermented foods
https://images.app.goo.gl/FzqyPWbLzsVhGVnSA

Enzymes
Enzymes are biological catalysts that speed up the rate of the biochemical
reaction.
Most enzymes are three dimensional globular proteins (tertiary and quaternary
structure).
Enzymes are specific
Enzymes are catalyst as one enzyme can perform the same job over & over again
millions of times without being consumed
Enzymes are efficient
Enzymes are proteins

Enzymes
The active site of an enzyme is the region that binds substrates, co-factors and
prosthetic groups and contains residue that helps to hold the substrate.
Active sites generally occupy less than 5% of the total surface area of enzyme.
Active site has a specific shape due to tertiary structure of protein.
A change in the shape of protein affects the shape of active site and function of
the enzyme.
STRUCTURE OF ENZYMES

Enzymes
ACTIVE SITE
Active site can be further divided into:
Active Site
Binding Site Catalytic Site

Enzymes
 Co-factor is the non-protein molecule which carries out chemical reactions that
cannot be performed by amino acids.
 Co-factors are of two types:
 Organic co-factors
 Inorganic cofactors
https://images.app.goo.gl/JfuLMTLqpomPxoMg8

Enzymes
INTRACELLULAR AND EXTRACELLULAR ENZYMES
 Intracellular enzymes are synthesized and retained in the cell for the use of cell
itself
They are found in the cytoplasm, nucleus, mitochondria and chloroplast.
Example : Oxidoreductase catalyses biological oxidation.
Enzymes involved in reduction in the mitochondria
 Extracellular enzymes are synthesized in the cell but secreted from the cell to
work externally
Example : Digestive enzyme produced by the pancreas, are not used by the cells in
the pancreas but are transported to the duodenum.

Enzymes
CHARACTERISTICS
 Enzymes speed up the reaction by lowering the activation energy of the
reaction.
 Their presence does not affect the nature and properties of end product.
 They are highly specific in their action that is each enzyme can catalyze one
kind of substrate.
 Small amount of enzymes can accelerate chemical reactions.
 Enzymes are sensitive to change in pH, temperature and substrate
concentration.
 Turnover number is defined as the number of substrate molecules
transformed per minute by one enzyme molecule

Enzymes
Classification of enzymes
ENZYME CLASS REACTION TYPE EXAMPLES
Oxidoreductases Reduction-oxidation (redox) Lactate dehydrogenase
Transferases Move chemical group Hexokinase
Hydrolases
Hydrolysis; bond cleavage with transfer of
functional group of water
Lysozyme
Lyases Non-hydrolytic bond cleavage Fumarase
Isomerases Intramolecular group transfer (isomerization)
Triose
phosphate isomerase
Ligases
Synthesis of new covalent bond between
substrates, using ATP hydrolysis
RNA polymerase

Enzymes
LOCK AND KEY MODEL
 Proposed by EMIL FISCHER in 1894
 Lock and key hypothesis assumes the active site of an enzymes are rigid in its
shape
 There is no change in the active site before and after a chemical reaction
https://images.app.goo.gl/iR1Udy4z9R4uqsDR7

Enzymes
INDUCED FIT MODEL
 More recent studies have revealed that the process is much more likely to involve an induced fit
model (proposed by DANIAL KOSH LAND in 1958)
 According to this exposure of an enzyme to substrate cause a change in enzyme, which causes
the active site to change its shape to allow enzyme and substrate to bind.
https://images.app.goo.gl/iR1Udy4z9R4uqsDR7

Enzymes
Background
• For many thousand of years , man has used Naturally occurring microorganisms
(bacteria, yeast, mold &the enzymes they produced to make food such as
: “Bread, Cheese, beer& Wine”
• Example: in bread making , amylase is used to breakdown flour into soluble
sugars,which are transformed by yeast into : Alcohol & carbon dioxide,
• This make bread rise.

Enzymes
• Today, enzymes are used for an increasing range of application:
• Bakery, cheese making, starch processing , tenderizing of meat, production of fruit
juice& other drink
• Here they can improve texture, appearance, nutritional value & may generate
desirable flavor and aroma

Enzymes
Forms of enzymes
 Liquids
 Granules
 Capsules
 Immobilized preparation

Enzymes
Uses of enzymes
Production of bulk product such as glucose &
fructose
In food processing & food analysis
In laundry & automatic dishwashing detergent
The textile, pulp & paper & animal feed industries
Clinical diagnosis & Therapy
Genetic engineering

Enzymes
Sources
Enzymes used in food industry can be derived from 3 sources:
1- Animal
2- Vegetable
3- Microbial

Enzymes
Animal derived enzymes
-There are 3 enzymes commonly used in food preparation that are derived from animal
tissue
1-rennet: an extract of fourth stomach of calf & is rich in rennin & pepsin
-Both of these are protease that cause milk to curdle into cheese.
2-Lipase : used to impart buttery flavor to oils by degrading some of lipids& to hasten the aging of
cheese.
3- Pancreatin: is rich in aplethora of protease.
-is used to modify protein to make it more easily digested.
Trypsin : used for the same purpose.
*Pancreatic tissue & its derivatives are the ingredient of infant food formulas to break down the protein
for children who cannot digest it.

Enzymes
Plant derived enzymes
• There are 3 major plant-derived protease used commercially today :
-Papain is derived from papaya plant.
-Bromelain from the pineapple plant.
-Ficin from the fig papain& bromelain which are commonly used as meat
tenderizers.
Ficin being more limited in use due to its more dangerous proteolytic activity.
-Barley amylase is also used to make maltose syrup.

Enzymes
Microbial enzymes
 The recent explosion of interest in enzymes involve the 3rd source (Microbial)
 The growth of microorganisms on nutrient media allow these microbes to
produce varied enzymes as a part of their natural metabolic function.
 If the growth of microorganisms can be manipulated in such away so that the
microorganism produced desired enzyme, these enzymes can be harvested &
concentrated for use in other application
 This is the heart of enzyme production via fermentation

Enzymes
Meat Tenderizing Enzymes
 The two most often used meat tenderizing enzymes are Papain and
Bromelain.
 Both are derived from plant sources. These are the papaya fruit and
the pineapple plant.
 Other sources of enzymes have been cited for meat
tenderization such as B. subtilis; A. oryzae & pancreas

Probiotics and their benefits were
discussed
Enzymes, types and applications were
studied
Week 10-Lesson 1 Summary
Topic 1
Probiotics and Enzymes

Course Progress-Week 10
Lesson 1. Probiotics and Enzymes
Lesson 2. Biofertilizers
Lesson 3. Biomaterials and Bioenergy

Bio-fertilizers
 Biofertilizers are the compounds that enrich the nutrient quality of the soil by using
microorganisms that establish symbiotic relationships with the plants
 These are the microbial inoculants which are artificially multiplied cultures of certain soil
microorganisms that can improve soil fertility and crop productivity
 Biofertilizers add nutrients through their activities like nitrogen fixation, phosphorus
solubilization and stimulating plant growth through the synthesis of growth promoting
substances.

Bio-fertilizers
 Plant growth promoting microorganisms include bacteria such as Azospirillum
spp., Pseudomonas spp., Bacillus spp. while fungus include Trichoderma spp.
 Biofertilizers include organic fertilizers which are rendered in an available form
due to interactions of microorganisms or their association with plants majorly
Rhizobium spp.

Bio-fertilizers

Bio-fertilizers
• Majorly, biofertilizers include the following types:
• Rhizobium spp. as symbiotic nitrogen fixers
• Azospirillum spp. and Azotobacter spp. Asymbiotic nitrogen fixers
• Algae biofertilizers
• Phosphate solubilizing bacteria
• Mycorrhizae

Bio-fertilizers
https://images.app.goo.gl/jgqxRWfnXrRJvjKJ9

Bio-fertilizers
 Rhizobium is a Gram negative bacterium which inhabits the root nodules of most of the
leguminous plants
 Rhizobia are soil inhabiting bacteria that fix nitrogen after becoming established inside the
root nodules
 Rhizobia donot produce spores and are aerobic and motile too
 Rhizobia maintain symbiotic relationships with legumes by responding chemotactically to
flavonoid molecules released as signals by the legume host plant.
Rhizobium

Bio-fertilizers
There are some steps involved in mass production of Rhizobium to use them as
biofertilizers.
These are as follows:
 Preparation of mother or starter culture
 Preparation of broth culture
 Preparation of carrier
 Preparation of inoculate (Mixing)
 Maturation
 Filling and packaging
 Quality checking
 Storage

Bio-fertilizers
 Cyanobacteria which are also known as blue-green algae, are photoautotrophic and
prokaryotic in nature
 They are free living and fix the atmospheric nitrogen in moist soils
 They also include unicellular as well as filamentous species having specialized cells known as
heterocysts such as Anabaena and Nostoc
 These cells are the site for nitrogen fixation and few of those which are non heterocystous can
reduce N2 into NH3 i.e nitrogen fixation.

Bio-fertilizers
The mass production of BGAcan be processed in the following ways mainly:
1. Trough method
2. Pit method
3. Field method

Bio-fertilizers
 This method is basically used in laboratory where zinc and iron troughs are used
 These are dimensionally 2 x 3 cm in width and 22 cm in height
 Trough is filled with about 10kg of soil and 200g of superphosphate is spread on it
 Water is poured upto 5-10cm height and calcium carbonate is added to adjust pH around 7
 Then, starter culture is sprinkled over it
 Trough is kept in sunlight where BGAis developed
 Watered everyday
 After sufficient growth of BGA, soil is allowed to dry and the dry flakes are collected and
packed for algalization
Trough Method

Bio-fertilizers
 In this method, under full sunlight, shallow pits are maintained
 To avoid percolation, polythene sheets are lined inside the pit
 Soil is filled in pit upto 20 cm and watered for 10 cm height
 Then, carbofuran and saw dust are added along with the starter culture sprinkling
 Then, similarly after the growth of BGA, the dry flakes are collected and packed.
Pit Method

Different types of bio-fertilizer were
studied
Mass production and its applications were
discussed
Week 10 Lesson 2 Summary
Topic 2
Biofertilizer

Bio-materials
• A biomaterial is a nonviable material used in a medical device,
intended to interact with biological systems.
• Defined by their application NOT chemical make-up.

Bio-materials
 Physical Requirements
 Hard Materials.
 Flexible Material.
 Chemical Requirements
 Must not react with any tissue in the body.
 Must be non-toxic to the body.
 Long-term replacement must not be biodegradable.

Bio-materials
 Grow cells in culture.
 Apparatus for handling proteins in the laboratory.
 Devices to regulate fertility in cattle.
 Aquaculture of oysters.
 Cell-silicon “Biochip”.

Bio-materials
https://images.app.goo.gl/x9iFJKXiRn7iZGjB7

Bio-materials
First Generation Biomaterials
 Specified by physicians using common and borrowed materials.
 Most successes were accidental rather than by design.

Bio-materials
Second Generation of Biomaterials
 Developed through collaborations of physicians and engineers.
 Engineered implants using common and borrowed materials.
 Built on first generation experiences.
 Used advances in materials science (from other fields).

Bio-materials
Third generation implants
 Bioengineered implants using bioengineered materials.
 Few examples on the market.
 Some modified and new polymeric devices.
 Many under development.

Bio-materials
Examples of Biomaterial Applications
 Heart Valve
 Artificial Tissue
 Dental Implants
 Intraocular Lenses
 Vascular Grafts
 Hip Replacements

Bio-materials
Hostreactionstobiomaterials
■ Thrombosis
■ Hemolysis
■ Inflammation
■ InfectionandSterilization
■ Carcinogenesis
■ Hypersensitivity
■ SystemicEffects

Bio-materials
 Tomorecloselyreplicate complextissue architectureandarrangementin vitro.
 Tobetterunderstandextracellularand intracellularmodulators ofcellfunction.
 To develop novel materials and processing techniques that are compatible with biological
interfaces.
 Tofindbetterstrategies forimmune acceptance.
Challenges

Bio-materials
Biomaterials - Industry
 Nextgenerationofmedicalim
plantsand therapeuticmodalities.
 Interface of biotechnologyandtraditional engineering.
 Significantindustrialgrowthinthenext15years--potentialofamulti-billiondollar industry.

Bio-materials
 Baxter International develops technologies related to the blood and circulatory system.
 Biocompatibles Ltd. develops commercial applications for technology in the field of biocompatibility.
 Carmeda makes a biologically active surface that interacts with and supports the bodys own control
mechanisms
 Collagen Aesthetics Inc. bovine and human placental sourced collagens, recombinant collagens, and
PEG-polymers
 Endura-Tec Systems Corp. bio-mechanical endurance testing ofstents, grafts, and cardiovascular
materials
 Howmedica develops and manufactures products in orthopaedics.
 MATECH Biomedical Technologies, development of biomaterials by chemical polymerization
methods.

Bio-materials
 Medtronic, Inc. is a medical technology company specializing in implantable and invasive therapies.
 Molecular Geodesics Inc., biomimetic materials for biomedical, industrial, and military applications
 Polymer Technology Group is involved in the synthesis, characterization, and manufacture of new
polymer products.
 SurModics, offers PhotoLink(R) surface modification technology that can be used to immobilize
biomolecules
 W.L. Gore Medical Products Division, PTFE microstructures configured to exclude or accept
tissue ingrowth.
 Zimmer, design, manufacture and distribution of orthopaedic implants and related equipment and
supplies

Bio-energy
• Bioenergy is energy derived from biomass and biogas source
• It is a renewable energy source
• BIOMASS
• Biomass is organic matter from plants, micro- -organism grown on land and water and their
derivatives.
• The energy obtain from biomass is also called the biomass energy.
• It is a renewable energy source.
• Because organic matter generated everyday.
• Coal, petroleum, natural gas are not come in biomass category because they produce from dead,
buried biomass under high pressure and temperature during several millions of year.

Bio-energy

Bio-energy
Type of biomass
Biomass are classified in three group
1. Biomass from cultivated like fields, crops, forests etc.
2. Biomass derived from wastes like municipal waste, animal dung etc.
3. Biomass converted into liquid fuels.
• In first group the biomass is directly converted into energy by burning the biomass.
• Second group the biomass is fermented anaerobically to obtain gaseous fuel like bio-gas.

Bio-energy
 Naturally occurring bacteria breakdown organic material (such as agricultural energy
crops like Giant King Grass) in the absence of oxygen resulting in the creation of
methane and carbon dioxide, which make up the composition of biogas
 This process is called anaerobic digestion and occurs in large enclosed tanks
 The biogas is collected from the anaerobic digestion tanks and processed through
a generator to produce renewable electricity.
 In third group biomass is converted into ethanol and methanol to use in a liquid fuels
in engine.

Bio-energy
Biomass conversion processes
1. Direct combustion
2. Thermochemical conversion
3. Biochemical conversion direct combustion
• Combustion is the oldest and most frequently applied process to extract the energy content from
solid biomass.
• During combustion, most of the energy is released in form of heat.
• Different thermodynamic processes can be used to transform part of this heat into electric power.

Bio-energy
Thermochemical conversion
 Thermochemical conversion the process convert the biomass and its residues to
fuel, chemicals and power using gasification heating of biomass with about one
third of oxygen is necessary for complete combustion produce mixture of CO2 and
hydrogen known as syngas
 Pyrolysis heating biomass in absence or produce a liquid pyrolysis oil
 They both are use as fuel

Bio-energy
Biochemical conversion
 Biochemical conversion by micro-organic biomass to biofuel are slow process
taking place low temperature
 The principle conversion process is fermentation
 Fermentation is a process of decomposition of organic matter by micro-organism
 Example fermentation, decomposition of sugar to form ethanol and carbon
dioxide by yeast and ethanol forming acetic acid in making vinegar

Bio-energy
Application of biomass
 generating electricity
 the producer gas from the biomass gasifier is first cleaned and cooled and then
used as a fuel in an IC engine.
 Biomass gasifier plants in an industry or an institute are usually used as captive
power generation unit
 In India, a large number of systems have been put-up in rice mills, with rice-husk
as the feed material for gasifiers

Bio-energy
Advantage of biomass
 Biomass Energy (or Bioenergy) is a renewable energy
 Biomass is always available it can be found anywhere and includes organic matter such as plants,
animals or waste products from organic sources
 Less pollution is generated
 Reduce Fossil Fuel dependency
 With the majority of homes and businesses using oil to provide energy, oil will gradually run out if
people do not switch to a renewable energy source such as biomass

Bio-energy
Advantage of biomass
 The use of biomass will therefore reduce the dependency on fossil fuels
 Clean energy
 As biomass is relatively clean, it can be used in such commercial businesses as
airlines, meaning it is good for the environment and good for businesses

Bio-energy
Disadvantage of biomass
 The initial costs of a biomass is high
 Harmful to the environment: Although there is a large reduction of carbon dioxide emissions
compared to other systems, there is an increase in methane gases, which can also be harmful to
the Earth’s ozone layer
 Consumes more fuel

Bio-energy
U.S. Department of Energy Bioenergy Research Centers: 2020

Types of biomaterials and thir applications
were discussed
Bioenergy, biomass and their conversion
were studied
Week 10 Lesson 3 Summary
Topic 3
Biomaterials and Bioenergy

Course progress
Week 11
•Lesson 1: Wastewater treatment
•Lesson 2: Role of genetic
engineering: Insulin production
•Lesson 3:Antibiotics

Week 11-Lesson 1 Topics
Topic 1. An introduction to Wastewater treatment
Topic 2. Physical processes -- Clarification/Settling
Topic 3. Chemical processes: Precipitation and clarification
Topic 4. Biological wastewater treatment
Topic 5. Other wastewater treatment processes

© Kalasalingam academy of research and education COURSE NAME: BIOLOGY FOR ENGINEERS
Wastewater treatment
What is wastewater treatment?
 Usually refer to sewage treatment, or
domestic wastewater treatment.
 Process of removing contaminants
from wastewater, both runoff and
domestic to produce water that is
safe for environment.
•Physical treatment
•Biological treatment
•Chemical treatment
The main objectives of the conventional wastewater
treatment processes are the reduction in biochemical
oxygen demand, suspended solids and pathogenic
organisms.

Applicability and Selection of Methods
 Different processes are used to treat
wastewater depending on the contaminants
present
 Similarly, different processes are used to treat
sludge, depending on the objective of treatment
Wastewater Composition
Solids: density, particle size, level of Volatile
Suspended Solids
Biochemical Oxygen Demand
Temperature
Ammonia
Nutrient levels
Wastewater treatment plant

The characteristics of untreated wastewater of some major industries
Industry Wastewater characteristics
Iron and steel mills Suspended solids, sulphides, cyanide, thiocyanate, oxides of Cu, Cd, Hg,
Oil, cyanogens, cyanate, phenols, naphtha, acids, alkali, iron salts, coke,
limestone etc.
Tanneries Chromium, calcium, high salt content, colour, dissolved and suspended ,
matter etc.
Distilleries Very high COD, low pH, high organic matter, high suspended and
dissolved solids containing nitrogen, high potassium etc.
Thermal power plants Heat, heavy metals, dissolved solids, inorganic compounds etc.
Pulp and paper Suspended solids, sulphides, sulphites, bleaching agents, colour, high or
low pH, carbohydrates (cellulose fibres, wood, bark, sugars), organic
acids, BOD, COD, high temperature, dissolved substances etc.

The characteristics of untreated wastewater of textile industries

• Screening
• Sedimentation
• Filtration
Physical
Processes
• Precipitation
• Chlorination
• Disinfection
Chemical
Processes
• Aerobic
• Anaerobic
• Attached or Suspended
Biological
Processes
The purpose of industrial water
treatment is to remove impurities from
the source water. There are a number
of methods to achieve this including:
biological processes, physical
equipment, and chemical treatment.

Physical Processes -- Clarification/Settling
Screen,
Bag,
Cartridge or
Similar
Filters
Multi-
Media
Depth
Filters
Membrane
Filters
Purpose of physical treatment
 Influx (Influent)
 Removal of large objects (Ex: sticks, rags, toilet paper, tampons)
 Removal of sand and grit
 Fats, oils, and greases
 Larger settable solids including human waste, and floating materials
 Produce a homologous liquid for later biological treatment

Chemical Processes
Chemical coagulation is the process of adding a chemical
which can destabilize the colloidal and suspended
particles in the wastewater.
As a result of destabilization the size of particles will
increase and they settle as floc due to flocculation and
agglomeration.
These settled flocs can be removed using a
sedimentation tank and the supernatant will be the treated
effluent.
Typical coagulants used are natural and synthetic organic
polymers, metal salts such as Alum, Ferric sulfate etc
Coagulation & Clarification
Industrial Clarifier

Chemical Processes
Industrial precipitation
• Phosphorus is presented in wastewater is
generally seen in the form of organic
phosphate, polyphosphate or
orthophosphate.
• This reaction consisting of adding
chemicals such as alum, ferric chloride
into the wastewater and they coagulate
the phosphorous presented in it.
• The coagulated material will precipitate
out such as Aluminum phosphate and
ferric and this can be clarified further.
Chemical precipitation

Biological wastewater treatment
 Biological treatment involves use of
microorganisms such as bacteria,
viruses and protozoa
 They metabolize the biological content
(dissolved organic matter) of the
sewage
 The contaminants of organic
substances are digested as food along
with other energy source by the cell

Biological wastewater
treatment
Suspended and
attached
treatment
Suspended
growth
Attached
growth
Aerobic and
Anaerobic
Aerobic Anaerobic
The suspended growth as in conventional
activated sludge is a nominal process applied
as a biological treatment in a water treatment
plant. On the other hand, attached growth
processes can be obtained by combining
biofilm carriers and activated sludge in one
treatment step.
Attached growth
Suspended growth
Microbes

AEROBIC DIGESTION ANAEROBIC DIGESTION
Need oxygen Do not require oxygen
Convert into carbon
dioxide (co2)
Produce biogas, which
can be used in
generators for electricity
High energy is required Energy is produced in the
form of methane
Excellent effluent quality
in terms of BOD, COD
and nutrient removal is
achieved
Effluent quality in terms
of COD is fair, further
treatment is required.

Fixed film systems
• Activated sludge
Trickling filters
• Trickling biofilters/biological filters
Rotating biological contactors
Three Approaches
• SUSPENDED FILM SYSTEMS
• Stir and suspend microorganisms
in wastewater
• Settled out as a sludge
• Pumped back into the incoming
wastewater
• Ex: activated sludge, extended
aeration

Activated sludge process
Primary wastewater mixed with bacteria
rich (activated) sludge and air or oxygen is
pumped into the mixture
Mixed community of microorganisms
Both aerobic and anaerobic bacteria make
up about 95% of the activated sludge
biomass.
They grow in wastewater by consuming
biodegradable materials such as proteins,
carbohydrates, fats and similar compounds.
COMPONENTS OF ACTIVATED
SLUDGE PROCESS
AERATION TANK
oxygen is introduced into the system
AERATION SOURCE
ensure that adequate oxygen is fed into the
tank and provided pure oxygen or
compressed air
CLARIFIER / SETTLER
activated-sludge solids separate from the
surrounding wastewater

Raw water
Air
Recycle Sludge
Clarifier/Settler
Treated water
Sludge
Sludge treatment
Aeration Tank
Aeration Tank

Real industrial activated sludge process Sludge Treatment
 Sludge are the product of
biological treatment of
wastewater
 Sludge comprise solids
found in wastewater plus
organisms used in the
treatment process
 Disposal is a major issue
various disposal
techniques are used but
each has advantages and
disadvantages

Trickling filters bed
Media made of coke (carbonized coal), limestone chips or
specially fabricated plastic media.
Microorganisms are attached to the media in the bed and
form a bio film over it spread wastewater over this bio film
of aerobic microorganisms that will oxidize the organic
matter

Rotating Biological Contactors
• Disc, biofilm, aerobic reactor
system
• Solid media encourages
microbial growth in a static bio
film
• Primary function is reduction
of organic matter

Membrane bioreactors
Improvement of the conventional activated
sludge process
Secondary Clarifier is replaced by A biological
aeration basin followed by membrane unit for
the separation of treated water from the mixed
solution in the bioreactor
Membrane Fibers have billions of microscopic
pores on the surface
The Pores form a barrier to impurities , while
allowing pure water molecules to pass
Water is drawn through the pores using gentle
suction
Membrane bioreactors

Chlorination UV light radiation Ozonation
Most common
Advantages:
Low cost & effective
Disadvantages:
chlorine residue could be
harmful to environment
Damage the genetic structure of
bacteria, viruses and other
pathogens.
Advantages:
No chemicals are used
water taste more natural
Disadvantages:
High maintenance of the UV-lamp
Oxidized most pathogenic
microorganisms
Advantages:
safer than chlorination
fewer disinfection by-product
Disadvantage:
high cost
Other wastewater treatment processes

Summary
 Wastewater treatment is a process used to remove contaminants from wastewater or sewage
and convert it into an effluent that can be returned to the water cycle with minimum impact on
the environment, or directly reused.
 Pollutants in municipal wastewater (households and small industries) are removed or broken
down.
 Since the microbes have a natural ability to degrade pollutants from wastewater.
 Thus, advanced technologies using microbes must be applied such as Advanced Oxidation
Processes (AOPs) and membrane-separation technologies, and perhaps their combined
application may constitute today, the best option for wastewater treatment and reuse schemes.

© Kalasalingam academy of research and education COURSE NAME: BIT18R371/BIOPROCESS PRINCIPLES
Course progress
Week 11
•Lesson 1: Wastewater treatment
•Lesson 2: Role of genetic
engineering: Insulin production
•Lesson 3:Antibiotics

Topic 1. An introduction to
Topic 2. Physical processes --
Clarification/Settling
Topic 3. Chemical processes:
Precipitation and clarification
Topic 4. Biological wastewater
treatment
Topic 5. Other wastewater treatment
processes
Week 11_Lesson 1 Summary
 Wastewater treatment is a process used to remove contaminants from
wastewater or sewage and convert it into an effluent that can be
returned to the water cycle with minimum impact on the environment, or
directly reused.
 Pollutants in municipal wastewater (households and small industries)
are removed or broken down.
 Since the microbes have a natural ability to degrade pollutants from
wastewater.
 Thus, advanced technologies using microbes must be applied such as
Advanced Oxidation Processes (AOPs) and membrane-separation
technologies, and perhaps their combined application may constitute
today, the best option for wastewater treatment and reuse schemes.

Topic 1. An introduction to Genetic engineering
Topic 2. Modern biotechnological applications: An interdisciplinary
challenge
Topic 3. Modern biotechnological applications: Covid 19 vaccine
Topic 4. An introduction to recombinant protein insulin
Topic 5. Role of genetic engineering in insulin production

Introduction to Genetic Engineering
Father of Genetic Engineering is Paul Berg.
He was the first person who developed
recombinant DNA technology.
The change in genetic make up of living cells
by inserting desired gene through a vector in
called genetic engineering (GE).

Role of Genetic Engineering: Insulin Production
The beginning
 The first genetically modified animal was mouse created in
1973 by Rudolf Jaenisch.
 In 1993, an antibiotic resistant gene was inserted in tobacco
plant, leading to first genetically modified plant.
 In 1978, the technology was commercialized with the
production of insulin.
 In 1994, first genetically modified food Tomato was made.

Role of Genetic Engineering
Important Terms
Gene: The gene is small piece of DNA that
encodes for a specific protein.
Recombinant DNA (rDNA): The DNA
formed by joining DNA segment of two
different organism.
Recombinant DNA technology: The
technique by which gene of interest is
transferred to the host.
Genetically modified organism: The
organism whose genetic make up is
altered/changed using rDNA technology.
Steps involved in genetic engineering
1. Isolation of desired DNA fragment(gene of
interest) with the help of restriction enzymes.
2. Isolation of DNA vector.
3. Construction of rDNA. In this gene of interest
is inserted into the vector.
4. Introduction of vector containing recombinant
into the host cell.
5. Multiplication of Host cells containing
recombinant DNA.
6. Expression of cloned gene.
7. Selection of Recombinant cells.

What is a recombinant protein?
 Recombinant proteins are proteins that are artificially made
through the recombinant DNA technology.
 Proteins can be used in many areas, such as diagnostic tools,
vaccines, therapeutics, detergents, cosmetics, food production,
and feed additives.
 The recombinant DNA technology provides a more efficient
method to obtain large amounts of proteins.
 For example, insulin, a hormone that acts as a key regulator of
blood sugar and is reduced in patients with diabetes, has
already been produced with the recombinant DNA technology,
which saves many lives.
 However, there are still concerns about the safety and ethics of
the use of recombinant DNA technology.
How are recombinant proteins
manufactured?
Using recombinant DNA technology,
scientists are able to create new DNA
sequences that would not naturally
exist under normal circumstances and
environmental conditions.

Requirements for recombinant protein production
Vector:
 In molecular cloning, a vector is a DNA molecule used as a
vehicle to artificially carry foreign genetic material into another
cell, where it can be replicated and/or expressed
 (e.g: plasmid, cosmid, Lambda phages).
Host:
 It is the cell where the recombinant DNA is allowed to multiply
to produce several copies. e.g. Bacteria, yeast etc.
 The host should be non-pathogenic, harmless micro-organism
which is easy for cultivation. The bacterium Escherichia coli is
the most commonly used host in recombinant DNA technology.

Modern biotechnological applications: An interdisciplinary
challenge
 Modern biotechnology" replacement of
conventional process into newer
techniques.
 For example: genetic engineering and
cell fusion from more conventional
methods such as breeding, or
fermentation.
 Most often the term "biotechnology" is
used interchangeably with "modern
biotechnology“.
Pentavalent: Five
vaccines in a single dose
Availability and Affordability
of foods and medicines are possible
with the wonders of biotechnology
Modern facility at Serum Institute for COVID 19 vaccine production

challenge
Modern Biotechnology covers such key topics as:
 Metabolic engineering
 Enzymes and enzyme kinetics
 Biocatalysts and other new bioproducts
 Cell fusion
 Genetic engineering, DNA, RNA, and genes
 Genomes and genomics
 Production of biopharmaceuticals
 Fermentation modeling and process analysis
Metabolic engineering

challenge

Modern biotechnological applications: Covid 19 vaccine
Now, both Pfizer and Moderna are testing their separate
vaccine candidates that use messenger RNA, or mRNA, to
trigger the immune system to produce protective antibodies
without using actual bits of the virus.

Modern biotechnological applications: Covid 19 vaccine
Promising yeast-expressed SARS-CoV-2 vaccine candidate effective in mice
Expression of viral
protein on yeast
Availability
and
Affordability

What is insulin? What is insulin?
Insulin is a hormone made by one of the body's organs called
the pancreas.
 Insulin helps your body turn blood sugar (glucose) into
energy.
 It also helps your body store it in your muscles, fat cells, and
liver to use later, when your body needs it.

Do you think that
bacteria can produce
human insulin
hormone?
Recombinant DNA technology

Pig Pig pancreas
Homogenisation Partial purification
Insulin
(less
doses)
Traditional method of insulin production
Recombinant E. coli
Available and Affordable
No allergic reactions
New industrial method of insulin
production

Is it possible to overcome
insulin needle phobia?
How do you keep your
insulin cold while
travelling?
Insulin
pills !!!!
Modern
Biotechnology

Commercial manufacture of a new recombinant-DNA derived Insulin
Biochemicals
Animal
Tissue
Microorganisms E.coli
DNA
Gene
Molecular
scissors
Plasmid Vector
Recombinant DNA Technology
Insertion
Plasmid multiplication
Gene Expression
Cell division
Culture
Bench top bioreactor
Pilot scale bioreactor
Industrial scale operation
Product recovery
Packaging
Marketing

Summary
1. Genetic engineering is the process of using recombinant DNA (rDNA) technology to alter the genetic makeup of an
organism.
2. Genetic engineering involves the direct manipulation of one or more genes. Most often, a gene from another species is
added to an organism's genome to give it a desired phenotype.
3. Type 1 diabetes is an autoimmune disease. It occurs when the insulin-producing islet cells in the pancreas are completely
destroyed, so the body can't produce any insulin. In type 2 diabetes, the islet cells are still working. However, the body is
resistant to insulin.
4. Insulin is naturally synthesized as pre-proinsulin in the pancreas. Proinsulin is composed of the amino acid chains that will
form insulin and a connecting 30 residue peptide, that joins one end of chain A to chain B.
5. Recombinant human insulin is produced predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use
in human.
6. However, there is an upmost need to increase the production by several fold of a biologically active insulin and its
analogues.

Week 11_Lesson 2 Summary
1. Genetic engineering is the process of using recombinant DNA (rDNA)
technology to alter the genetic makeup of an organism.
2. Genetic engineering involves the direct manipulation of one or more
genes. Most often, a gene from another species is added to an
organism's genome to give it a desired phenotype.
3. Type 1 diabetes is an autoimmune disease. It occurs when the
insulin-producing islet cells in the pancreas are completely destroyed,
so the body can't produce any insulin. In type 2 diabetes, the islet
cells are still working. However, the body is resistant to insulin.
4. Recombinant human insulin is produced predominantly using E. coli
and Saccharomyces cerevisiae for therapeutic use in human.
5. However, there is an upmost need to increase the production by
several fold of a biologically active insulin and its analogues.
Topic 1. An introduction to Genetic engineering
Topic 2. Modern biotechnological applications: An
interdisciplinary challenge
Topic 3. Modern biotechnological applications:
Covid 19 vaccine
Topic 4. An introduction to recombinant protein
insulin
Topic 5. Role of genetic engineering in insulin
production

Topic 1. An introduction to Biopharming
Topic 2. An introduction to antibiotics
Topic 3. Antibiotics: Classification
Topic 4. Mechanism of antibiotic resistance
Topic 5. General Principles of Antimicrobial Therapy

Biopharming
Biopharming / What is biopharming?
 Biopharming, also known as plant molecular farming,
refers to the use of genetically modified plants to
produce a wide range of pharmaceuticals and industrial
products.
 Plants such as tobacco, for example, can be genetically
engineered to produce therapeutic proteins, monoclonal
antibodies and vaccines to treat cancer, inflammatory
diseases and other life-threatening or debilitating
conditions.
 These products are termed plant-made pharmaceuticals.
They belong to a class of pharmaceuticals known more
generally as “biologics” or “biopharmaceuticals,” as
they are derived from living organisms.
Biopharming

Antibiotics
What is an antibiotic?
Antibiotic” is from antibiosis, meaning against life.
Substances derived from a microorganism or
produced synthetically (Sulfonamides &
Quinolones) to kill or suppress the growth of
other microorganisms.
Antibiotics are classified by several ways:
1. On the basis of mechanism of action
2. On the basis of spectrum of activity
3. On the basis of mode of action
Image source: https://economictimes.indiatimes.com/news/politics-and-
nation/dispelling-myths-and-misconceptions-about-coronavirus/are-
antibiotics-effective-in-preventing-and-treating-the-new-
coronavirus/slideshow/74443452.cms

Antibiotics: Classification
On the basis of mode of action
Bacteriostatic antibiotics:
• Tetracycline
• Chloramphenicol
• Erythromycin
• Lincomycin
Bacteriocidal antibiotics:
• Cephalosporin
• Penicillin
• Erythromycin
• Aminoglycosies
• Cotrimoxazole
Bactericidal: Kills bacteria, reduces bacterial load
Bacteriostatic: Inhibit growth and reproduction of
bacteria
 All antibiotics require the immune system to work
properly
 Bactericidal appropriate in poor immunity
 Bacteriostatic require intact immune system
Bactericidal and Bacteriostatic

On the basis of spectrum of activity
Antimicrobial spectrum: the scope that a
drug kills or suppresses the growth of
microorganisms.
Narrow-spectrum: The drugs that only act on
one kind or one strain of bacteria
(Ex.Isoniazide).
Broad-spectrum: The drugs that have a wide
antimicrobial scope.
(Ex.Tetracycline & Chloramphenicol)
Classification on the basis of spectrum of activity

1. Inhibition of cell wall synthesis:
• Penicillins, Cephalosporins, Bacitracin & Vancomycin
2. Inhibition of functions of cellular membrane:
• Polymyxins
3. Inhibition of protein synthesis:
• Chloramphenicol, Macrolides & Clindamycin
• Tetracyclines & Aminoglycosides
4. Inhibition of nucleic acid synthesis:
• Quinolones
• Rifampin
5. Inhibition of folic acid synthesis:
• Sulfonamides & trimethoprim
On the basis of mode of action
Classification on the basis of mode of action

Antibiotics: β lactams
β-Lactam Ring Thiazolidine Ring
β-Lactam antibiotics are bactericidal
agents that interrupt bacterial cell-wall
formation as a result of covalent binding to
essential penicillin-binding proteins (PBPs),
enzymes that are involved in the terminal
steps of peptidoglycan cross-linking in both
Gram-negative and Gram-positive bacteria.

Antibiotics: β lactams
Spectrum of Activity
 Very wide
 Gram positive and negative bacteria
 Anaerobes
 Spectrum of activity depends on the agent and/or
its group
Adverse Effects
 Penicillin hypersensitivity – 0.4% to 10 %
 Mild: rash
 Severe: anaphylaxis & death
 There is cross-reactivity among all Penicillins
 Penicillins and cephalosporins ~5-15%

Antibiotics: Aminoglycosides
 Inhibit bacterial protein synthesis by
irreversibly binding to 30S ribosomal unit
 Causes mRNA decoding errors
30S Ribosomal Unit Blockage by Aminoglycosides
 Aminoglycosides are potent, broad-
spectrum antibiotics that act through inhibition of
protein synthesis.
 The class has been a cornerstone of antibacterial
chemotherapy since streptomycin was first isolated
from Streptomyces griseus and introduced into
clinical use in 1944.

Antibiotics: Aminoglycosides
 Gram-Negative Aerobes
 Enterobacteriaceae; E. coli, Proteus sp.,
Enterobacter sp. Pseudomonas aeruginosa
 Gram-Positive Aerobes (Usually in
combination with ß-lactams)
 S. aureus and coagulase-negative
staphylococci
 Viridans streptococci
 Enterococcus sp. (gentamicin)
Adverse Effects
 Nephrotoxicity: Direct proximal tubular damage -
reversible if caught early
 Risk factors: High troughs, prolonged duration of
therapy, underlying renal dysfunction, concomitant
nephrotoxins
 Ototoxicity: 8th cranial nerve damage – irreversible
vestibular and auditory toxicity
 Vestibular: dizziness, vertigo, ataxia
 Auditory: tinnitus, decreased hearing
 Neuromuscular paralysis
 Can occur after rapid IV infusion especially with;
 Myasthenia gravis
 Concurrent use of succinylcholine during anaesthesia

Antibiotics: Macrolides
Mechanism of Action
 Bacteriostatic- usually
 Inhibit bacterial RNA-dependent protein synthesis
 Bind reversibly to the 23S ribosomal RNA of the 50S
ribosomal subunits
 Block translocation reaction of the polypeptide chain
elongation
 The macrolides are bacteriostatic antibiotics with a
broad spectrum of activity against many gram-positive
bacteria.
 Currently available macrolides are well tolerated,
orally available and widely used to treat mild-to-
moderate infections.
 Several macrolide antibiotics have been linked to
liver injury.

Antibiotics: Macrolides
Spectrum of activity
Gram-Positive Aerobes:
 Activity: Clarithromycin>Erythromycin>Azithromycin
 MSSA
 S. Pneumoniae
 Beta haemolytic streptococci and viridans streptococci
Gram-Negative Aerobes:
 Activity:Azithromycin>Clarithromycin>Erythromycin
 H. influenzae, M. catarrhalis, Neisseria sp.
 NO activity against Enterobacteriaceae
Anaerobes: upper airway anaerobes and atypical bacteria

Antibiotics: Fluoroquinolones
Mechanism of Action
Prevent:
 Relaxation of supercoiled DNA before
replication
 DNA recombination
 DNA repair
Gram-positive
Gram-Negative (Enterobacteriaceae H.
influenzae, Neisseria sp. Pseudomonas
aeruginosa)
 Ciprofloxacin is most active
 Atypical bacteria: all have excellent activity
 The fluoroquinolones are a family of broad spectrum, systemic
antibacterial agents that have been used widely as therapy of
respiratory and urinary tract infections.
 Fluoroquinolones are active against a wide range of aerobic
Gram-positive and gram-negative organisms.

Antibiotics: Tetracyclines
All have similar activities
Gram positives aerobic cocci and rods
Staphylococci
Streptococci
Gram negative aerobic bacteria
Atypical organisms
Mycoplasmas
Chlamydiae
Rickettsiae
Protozoa
Tetracycline: Short acting
Doxycycline: Long acting
Mechanism of action
 Inhibit protein synthesis
 Bind reversibly to bacterial 30S
ribosomal subunits
 Prevents polypeptide synthesis
 Bacteriostatic
Adverse Effects
 Oesophageal ulceration
 Photosensitivity reaction
 Incorporate into foetal and children
bone and teeth

Antibiotics: Glycopeptide
Vancomycin
 Vancomycin is an antibiotic medication used to
treat a number of bacterial infections.
 It is recommended intravenously as a treatment for
complicated skin infections, bloodstream infections,
endocarditis, bone and joint infections, and
meningitis caused by methicillin-resistant
Staphylococcus aureus.
E.g. Vancomycin and Teicoplanin
Mechanism of Action
 Inhibit peptidoglycan synthesis in the bacterial
cell wall
 Prevents cross linkage of peptidoglycan chains

Antibiotics: Metronidazole
 Metronidazole belongs to the nitroimidazole class of
antibiotics and is active against protozoa in addition to
anaerobic bacteria.
 It is bactericidal to anaerobic organisms through formation of
free radicals that inhibit DNA synthesis and cause DNA
degradation.
 Antibiotic, Amoebicide and Anti-protozoal
 Trichomonas Vaginalis
Mechanisms of Action: Molecular reduction
Spectrum of Activity & Uses
 Anaerobes
 Bacterial Vaginosis
 Pelvic Inflammatory Disease
Bacterial Vaginosis

Mechanisms of Antibiotic Resistance
Alteration of the target site of the antibiotic
One of the most problematic antibiotic resistances worldwide,
methicillin resistance among Staphylococcus aureus
Enzyme inactivation of the antibiotic
β-lactam antibiotics (Penicillins & Cephalosporins) can be inactivated
by β-lactamases.
Active transport of the antibiotic out of the bacterial cell
Active transport of the antibiotic out of the bacterial cell (efflux pumps)
as removal of some antibiotics e.g.Tetracyclines, Macrolides &
Quinolones
Decreased permeability of the bacterial cell wall to the antibiotic
Alteration in the porin proteins that form channels in the cell
membrane
e.g. Resistance of Pseudomonas aeruginosa to a variety of Penicillins
& Cephalosporins
 Antibiotic resistance happens when
germs like bacteria and fungi develop the
ability to defeat the drugs designed to kill
them.
 That means the germs are not killed and
continue to grow.
 Infections caused by antibiotic-resistant
germs are difficult, and sometimes
impossible, to treat.

General Principles of Antimicrobial Therapy
 Identification of the infecting organism should precede
antimicrobial therapy when possible.
 The pathogenic microorganism susceptibility to antimicrobial
agents should be determined, if a suitable test exists.
 Factors that influence the choice of an antimicrobial agent or its
dosage for a patient include the age, renal & hepatic function,
pregnancy status and the site of infection, etc.
 Selection of Antimicrobial Agent
Empiric therapy - prior to identification of Organism
Critically ill patients
Organism’s susceptibility to the antibiotic
Patient factors - immune system, renal/hepatic function
Effect of site of infection on therapy - blood brain barrier
Safety of the agent
Cost of therapy
Ideal Antimicrobial Agent
1. Have highly selective toxicity to the
pathogenic microorganisms in
host body
2. Have no or less toxicity to the host
3. Low propensity for development of
resistance
4. Not induce hypersensitive
reactions in the host
5. Have rapid and extensive tissue
distribution
6. Be free of interactions with other
drugs
7. Be relatively inexpensive

Summary
 Antibiotics are drugs used to treat bacterial infections.
 They are ineffective against viral infections and most other infections.
 Antibiotics either kill microorganisms or stop them from reproducing, allowing the body's natural
defenses to eliminate them.
 Although doctors try to use antibiotics for specific bacterial infections, they sometimes start
antibiotics without waiting for tests that identify the specific bacteria.
 Bacteria can develop resistance to the effects of antibiotics.
 Antibiotics can have side effects, such as upset stomach, diarrhea, and, in women, vaginal yeast
infections.
 Some people are allergic to certain antibiotics.

References
1. Pelczar MJ, Chan ECS and Krieg NR - Microbiology - Tata McGraw Hill, India- 7th Edition- 2010
2. McCarty PL - Environmental biotechnology: principles and applications - Tata McGraw-Hill Education – 2012
3. Singh B, Gautam SK and Chauhan MS - Textbook of Biotechnology - Pearson Education - 2012 (1st Edition).
4. Ramadoss P - Animal Biotechnology: Recent Concepts and Developments - MJb Publishers - 2008 (1st Edition).

Unit 4 Week 12 Outline
Lesson 1. Vaccines
Lesson 2. Monoclonal antibodies
Lesson 3. Stem cell technology
Lesson 4. Self-healing concrete

Course Progress- Week 12
Lesson 1: Vaccines
Lesson 2: Monoclonal antibodies
Lesson 3: Stem cell Technology
Lesson 4: Self-healing concrete

Vaccines
First Lesson
Topic 1
Immunization and Types of
immunization
Topic 2
Vaccines and History of
vaccination
Topic 3
Types of vaccines and
Examples
Topic 4
How vaccines are made?
Topic 5
Case study

Immunization
Immunization enables the body to better defend itself against diseases caused by
certain bacteria or viruses.
There are two types of immunization:
Active immunization
Passive immunization

Types of Immunization
Active Immunization
Methods of acquisition include natural infection, vaccines,
and toxoids
Relatively permanent
Passive Immunization
Methods of acquisition include natural maternal antibodies,
antitoxins and immunoglobulins
Protection transferred from another person or animal

Vaccines
The word "vaccine" was created by Edward Jenner.
Comes from the Latin word vacca, meaning cow.
Vaccines are preparations that contain one of the following:
 Non-infectious fragments of bacteria or viruses
 A usually harmful substance (toxin) that is produced by a bacteria but has been modified to
be harmless—called a toxoid
 Weakened (attenuated), live whole organisms that do not cause illness
The body’s immune system responds to a vaccine and stimulates to produce
antibodies and white blood cells that recognize and attack the specific bacteria or
virus contained in the vaccine.
Whenever the person is exposed to the specific bacteria or virus, the body
automatically produces these antibodies and other substances to prevent or lessen
illness.
The process of giving a vaccine is called vaccination, more generally termed as
immunization.
17 May 1749 –
26 January 1823

A brief history of vaccination
The practice of immunisation dates back several hundreds of years.
17th century China-Buddhist monks drank snake venom to confer immunity to snake bite
and variolation (smearing of a skin tear with cowpox to confer immunity to smallpox)
British scientist Edward Jenner is considered the founder of vaccinology.
Beginning in 1760, Edward Jenner began experimenting with material from cowpox, an
infectious disease that primarily affected cows but could also produce a mild disease in
human.
Cowpox was first observed in milkmaids due to prolonged exposure to infected cows.
Jenner observed that these milkmaids had developed natural immunity to smallpox
In 1796, he inoculated an eight-year-old boy named James Phipps with material from a
cowpox patient.
Jenner observed that when Phipps was exposed to smallpox material, demonstrated
immunity to smallpox and he did not develop the disease.
In 1798, the first smallpox vaccine was developed.

Types of Vaccines
 Whole-Organism
– Live Attenuated Viral/Bacterial
– Inactivated Viral/Bacterial
 Subunit Vaccines
– Purified Macromolecules
Polysaccharide
– Recombinant Antigen
 Toxoid Vaccines
 Nucleic Acid Vaccines
 Synthetic Peptide

Types of vaccines
Live Attenuated Vaccines- produced by modifying a disease-producing virus or
bacterium in a laboratory.
The resulting vaccine organism retains the ability to replicate (grow) and produce
immunity, but usually does not cause illness.
Inactivated vaccines are produced by growing the bacterium or virus in culture
media, then inactivating it with heat and/or chemicals (usually formalin).
In the case of subunit vaccines, the organism is further treated to purify only those
components (polysaccharides/protein) to be included in the vaccine.

Types of vaccines
Toxoid Vaccines
Toxoid vaccines use a toxins made by the pathogen that causes a disease.
Protein-based toxin is rendered harmless and used as the antigen in the vaccine to elicit immunity.
Immune response is targeted to the toxin instead of the whole pathogen.
Purified toxins is suppressed or inactivated either by heat or with formaldehyde (while maintaining
immunogenicity) to form toxoids
Viral vector vaccines
Viral vector vaccines use a modified version of a different virus as a vector to deliver protection.
Several different viruses have been used as vectors, including influenza, vesicular stomatitis virus
(VSV), measles virus, and adenovirus, which causes the common cold.

Types of vaccines
Nucleic Acid Vaccines
Next generation/platform-based vaccines
The use of nucleic acid-based vectors (DNA or RNA) as an alternative to live-attenuated immunization is
a novel strategy now under development and evaluation.
DNA-based vaccines are composed of purified closed-circular plasmid DNA or non-replicating viral
vectors containing genes that encode viral antigens
Once the DNA enters the mammalian cell, the encoded antigens are expressed through normal cellular
transcription and translation mechanisms

Peptide vaccines
The antigen to which the immune system responds is a relatively small number
of amino acids or peptide.
A possible alternative approach to immunization -to identify the peptide
sequences that trigger a protective immune response and to use completely
synthetic versions of these as the vaccine substance.
Advanced clinical development-malaria, hepatitis C virus, influenza virus, and
HIV-1

© Kalasalingam academy of research and education COURSE NAME
Examples
• Hepatitis A
• Flu
• Polio
• Rabies
Inactivated
vaccines
• BCG-Tuberculosis
• Measles, mumps, rubella (MMR combined
vaccine)
• Rotavirus
• Smallpox
• Chickenpox
Live-attenuated
vaccines

© Kalasalingam academy of research and education COURSE NAME: BIT18R101-BIOLOGY FOR ENGINEERS
Examples
• Hib (Hemophilus influenza type b)
• Hepatitis B
• Human papillomavirus (HPV)
• Whooping cough
• Pneumococcal disease
• Meningococcal disease
Subunit,
recombinant,
polysaccharide, and
conjugate vaccines
• Diphtheria
• Tetanus
Toxoid vaccines

Examples
• Zika
• Flu
• HIV
Viral vector
vaccines
• Avian influenza
• H1N1 pandemic influenza
• Zika virus
Nucleic Acid
vaccines

How vaccines are made?
First step is the generation of the antigen used to induce an immune response
This includes the growth and harvesting of the pathogen itself (later for inactivation or
isolation of a subunit or generation of a recombinant protein derived from the
pathogen.
Viruses are grown in cell cultures
Bacterial pathogens are grown in bioreactors using optimized media and conditions
 Recombinant proteins are produced in cultures of bacteria/yeast/mammalian cells
Egg based vaccine production- Many viruses can be propagated in embryonated
chicken eggs but the method is now only used for Influenza viruses
Cell culture based vaccine production

Steps involved in vaccine production
Selecting
the strains
for
vaccine
production
Culturing the
microorganisms
Harvesting
&
Purification
of
microorgani
sms
Inactivation
and
splitting of
organism
Formulation
of vaccine
Quality
control and
lot release
Upstream
Processing
Downstream Processing

Egg based vaccine production
1
• Embryonated eggs to be used should be from closed,
specific-pathogen-free, healthy flocks.
2
• Monitored at regular intervals for Bacteria, Virus and
Mycoplasma.
3
• 5 to 14 days after fertilization, a hole is drilled in the shell and
• Virus injected into the site appropriate for its replication
• Yolk sac, chorioallantoic membrane, amniotic cavity, allantoic
cavity

Egg based vaccine production
The eggs are incubated at about 33°C for 2 to 3 days, candled
for viability and lack of contamination from the inoculation, and
then the allantoic fluid is harvested
After propagation, the virus is harvested.
Harvesting of virus requires extracting infected cells,
break down of cell walls, and then collecting the virus.
After treatment of the infected cell line, the virus is
released into the supernatant, and the cellular debris is
centrifuged away.
Following purification, the virus is inactivated through a
chemical process.

Cell culture based vaccine production
Mammalian
cell culture
Inoculation Harvest Bulk Purification
Packaging Labeling Inspection Filling Formulation
Virus

Formulation of vaccine
Apart from microorganisms or its part (antigen), a vaccine contain the following
components:
The vaccine is formulated by adding adjuvant, stabilizers, and preservatives.
Adjuvants- enhance vaccine immunogenicity- Example: Aluminium salts (Alum)
Preservatives and Antibiotics- Prevent bacterial or fungal contamination of vaccine
Examples: Thimerosal, neomycin, streptomycin, polymyxin B, chlortetracyline and
amphotericin B
Stabilizers-, Protects vaccines from adverse conditions such as freeze-drying or
heat, thereby maintaining a vaccine’s potency Examples: Albumin, Phenols, Glycine,
Gelatin, Monosodium glutamate (MSG)

Case study- SARS-CoV-2 vaccines
Vaccines for the prevention of coronavirus disease 2019 (COVID-19) caused by severe
acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Covishield
Covishield is originally developed by Oxford University-Astrazeneca and has been produced and
marketed as 'Covishield' by Pune-based vaccine conglomerate, Serum Institute of India
Use of a viral vector made using a weakened strain of the common cold virus (adenovirus), which
contains genetic material similar to that of SARS-COV-2.
Upon administration, the body's defences recognize the spike protein and prepare antibodies to evade
out the infection

Covaxin
India's first fully-developed and produced COVID-19 vaccine, made by Hyderabad-
based Bharat Biotech.
Made using an inactivated version of the virus- i.e., the vaccine inactivates the
virus's ability to replicate but sustains its life so that the immune system could
mount a sufficient response when it comes in contact or recognizes an attack on
the body in the future.

Pfizer-BioNTech COVID-19 Vaccine-mRNA Vaccine
mRNA – or messenger RNA – composed of nucleotides linked in a unique order to
convey genetic information for the cells to produce the proteins or antigens encoded
by the mRNA.
Once mRNA in a vaccine is inside the body’s cells, the cells use their genetic
machinery to translate the genetic information and produce the antigens encoded
by the mRNA vaccine.
The antigens are then displayed on the cell surface, where they are recognized by
the immune system stimulating immune response, including the production of
antibodies against the antigen

References
1. https://www.who.int/health-topics/vaccines-and-immunization#tab=tab_1
2. Centers for Disease Control and Prevention

Week 12-First Lesson Summary
 Vaccines help reduce the risk of certain illnesses by introducing dead or weakened versions of
disease-causing germs (bacteria or viruses) to the immune system.
 Vaccines protect vulnerable people in our community – such as very young children, the elderly, or
those who are too sick to be immunised.
 Some vaccines offer lifelong immunity.
 Immunisation greatly reduces the risk of getting a disease, which in turn reduces the risk of
complications.

Course Progress –Week 12
Lesson 1: Vaccines
Lesson 2: Monoclonal antibodies
Lesson 3: Stem cell Technology
Lesson 4: Self-healing concrete

Monoclonal antibodies
Second Lesson
Topic 1
Antibodies
Topic 2
Monoclonal Antibody
Topic 3
Polyclonal versus Monoclonal
antibodies
Topic 4
Production of Monoclonal
antibody
Topic 5
Applications of Monoclonal
Antibodies

Antibodies
Antibodies or immunoglobulin’s are protein molecules produced by a specialized group of
cells called B-lymphocytes (plasma cells) in mammals.
Antibodies are a part of the defense system to protect the body against the invading
foreign substances namely antigens.
Each antigen has specific antigen determinants (epitopes) located on it.
The antibodies have complementary determining regions (CDRs) which are mainly
responsible for the antibody specificity.
In response to an antigen (with several different epitopes), B-lymphocytes gear up and
produce many different antibodies.
These types of antibodies which can react with the same antigen are designated as
polyclonal antibodies.

Monoclonal Antibody
Monoclonal antibody (MAb) is a single type of antibody that is directed against a
specific antigenic determinant (epitope).
In the early years, animals were immunized against a specific antigen, B-
lymphocytes were isolated and cultured in vitro for producing MAbs.
This approach was not successful since culturing normal B-lymphocytes is
difficult, and the synthesis of MAb was short-lived and very limited.
In 1975, George Kohler and Cesar Milstein (Nobel Prize, 1984) achieved large
scale production of MAbs.
They could successfully hybridize antibody—producing B-lymphocytes with
myeloma cells in vitro and create a hybridoma.

Derived from different B Lymphocytes
cell lines
POLYCLONAL MONOCLONAL
Derived from a single B cell clone
Batch to Batch variation affecting Ab
reactivity & titre
mAb offer Reproducible, Predictable &
Potentially inexhaustible supply of Ab
with exquisite specificity
Enable the development of secure
immunoassay systems.
NOT Powerful tools for clinical
diagnostic tests
Polyclonal versus Monoclonal antibodies

Production of Monoclonal antibody
HYBRIDOMA TECHNOLOGY
Step 1: - Immunization of Mice & Selection of Mouse Donor for Generation of
Hybridoma cells
ANTIGEN ( Intact cell/
Whole cell membrane/
micro-organisms ) +
ADJUVANT
(emulsification)
Ab titre reached in Serum
Spleen removed (source of cells)

Step 2: - Screening of Mice for Antibody Production
After several weeks of immunization
Serum Antibody Titre Determined
(Technique: - ELISA / Flow cytometry)
Titre too low
BOOST
(Pure antigen)
Titre High
BOOST
(Pure antigen)

Step 3: - Preparation of Myeloma Cells
+ 8 - Azaguanine
Myeloma Cells
HGPRT-
High Viability & Rapid Growth
Myeloma Cells
Immortal Tumor of Lymphocytes

Normal cells are fused with cancerous cell Line Eg. myeloma,
lymphoma
Fusion is accomplished with PEG (polyethylene glycol)
New hybrid cell exhibits properties of both cell types
Unlimited growth
Secretes monoclonal antibody

Step 4: - Fusion of Myeloma cells with Spleen cells
FUSION
PEG
MYELOMA CELLS
SPLEEN CELLS
HYBRIDOMA CELLS
ELISA PLATE
Feeder Cells
Growth Medium
HAT Medium
1. Plating of Cells in HAT selective
Medium
2. Screening of Viable Hybridomas
Harvest monoclonal
antibodies

+
Myeloma HGPRT Deficient
and Ig Deficient
Plasma Cells from
Immunized Animal

Senescence
Can Use Salvage
Pathway, No Senescence

HAT Medium

Senescence

HAT Medium

Applications of Monoclonal Antibodies
Diagnostic Applications- Cancers, Hormonal disorders, Infectious diseases,
Biosensors
Therapeutic Applications
-Transplant rejection-Muronomab-CD3
-Cardiovascular disease-Abciximab
-Cancer- Rituximab
-Infectious Diseases- Palivizumab
-Inflammatory disease-Infliximab
Clinical Applications
- Targeting Agents in Therapy, Imaging the target
Protein Purification

Monoclonal antibodies provide higher specificity than
polyclonal antisera because they bind to a single epitope and
usually have high affinity.
Monoclonal antibodies are typically produced by culturing
antibody-secreting hybridomas derived from mice.
Second Lesson Summary

Stem Cell Technology
Third Lesson
Topic 1
Stem Cells
Topic 2
Classification of Stem cells
Topic 3
Regenerative Medicine and
Stem cells
Topic 4
Stem cell based therapy
Topic 5
Stem Cell Banking
Topic 6
Applications of Stem Cell
Technology

Stem Cells
Coined by William Sedgwick in 1886
Stem cells are rare, undifferentiated cells in an organism and are defined by their
properties of
(1) self-renewal, the ability to undergo numerous cycles of cell division while
maintaining an undifferentiated state; and
(2) potency, the ability to generate cells of many lineages.
Stem cells function in early development and in adult organisms to maintain and
repair tissue integrity.

CLASSIFICATION OF
STEM CELLS
“Toti”- Whole
“Pluri”- Many
“Multi”- Several

Classification of Stem cells
Based on their potency, stem cells can be classified according to a hierarchical order as”
 totipotent
 pluripotent
 multipotent stem cells
Totipotent stem cells of the early cleavage stages are able to generate an entire organism when
separated.
Further cell division and blastulation gives rise to the trophoblast, eventually forming the placenta,
and inner mass cells, destined to become the fetus.
Isolation of inner mass cells yields pluripotent embryonic stem cells capable of generating all three
embryonic germ layers: endoderm, ectoderm and mesoderm.
Adult stem cells- rare, exist among differentiated tissues in specialized niches, and function primarily
in tissue maintenance and repair.
Adult stem cells- multipotent, lineage-restricted cells, and are capable of generating a single germ
layer, often of a single organ system.

Embryonic stem cells
Embryos - Embryonic stem cells are obtained by harvesting living
embryos which are generally 5-7 days old.
The removal of embryonic stem cells invariably results in the
destruction of the embryo.
Embryonic stem cells are derived from embryos that develop from
eggs that have been fertilized in vitro.
Fetuses - Another kind of stem cell, called an embryonic germ cell,
can be obtained from either miscarriages or aborted fetuses.
Embryos
Embryonic stem cells

How Human Embryonic Stem Cells are Derived?
Source:
https://stemcells.nih.gov/info/Regenerative_Medicine/2
006Chapter1.htm

Programming multipotent adult stem cells to pluripotent state
Cells from patient (skin/fibroblasts);
Grown in a dish
Treat cells with “reprogramming
factors”
Culture for few weeks
Pluripotent stem cells
Change culture conditions to stimulate cells to
differentiate into a variety of cell types
Blood cells Gut cells Cardiac muscle cells

Adult Stem cells
Sources:
Umbilical Cords, Placentas and Amniotic Fluid
Adult Tissues - bone marrow, peripheral blood, brain, spinal cord, dental pulp,
blood vessels, skeletal muscle, epithelia of the skin and digestive system, cornea,
retina, liver, and pancreas
Cadavers - Neural stem cells have been removed from specific areas in post-
mortem human brains as late as 20 hours following death.

Umbilical cord stem cells
Placenta and umbilical cord that are left over after birth is
a rich source of hematopoietic stem cells.
Umbilical cord stem cells -able to differentiate into bone
cells and neurons, as well as the cells lining the inside of
blood vessels.
Used to treat 70 different diseases, including leukemia,
lymphoma, and inherited diseases
Detached umbilical cord

Regenerative Medicine and Stem cells
Differentiated into
• Cells
• Tissue
• Organ
Tissue regeneration includes delivering specific types of cells or cell products to injured tissues or organs for
restoration of tissue and organ function.
Regenerative medicine- Process of replacing, engineering or regenerating human cells, tissues or organs to
restore or establish normal function
Stem cell therapy provides a new paradigm in tissue regeneration
The hematopoietic stem cell (HSC) is the best-studied and well-characterized multipotent stem cell.
It resides in the adult bone marrow niche
Able to regenerate all the cellular components of the blood.
For these reasons, HSCs represent an attractive target for regenerative medicine.

Differentiation capacity of HSCs has been heavily utilized in regenerative medicine and other stem
cell-based therapies.
CD34+ HSCs can be collected from the bone marrow, umbilical cord blood, or from peripheral
blood following granulocyte colony stimulating factor (G-CSF)-mobilization from the bone marrow.
 Transplantation of HSCs has become the standard treatment for numerous hereditary diseases
and malignant blood disorders
Possibility to regenerate all the cellular components of the blood system and to permanently
restore a functioning immune system damaged by natural or acquired conditions.

A potential advantage of using stem cells from an adult is that the patient's own
cells could be expanded in culture and then reintroduced into the patient.
The use of the patient's own adult stem cells would mean that the cells would not
be rejected by the immune system.
Embryonic stem cells from a donor introduced into a patient could cause
transplant rejection.

Is Stem Cell Research Ethical?
Embryonic Stem Cells - always morally objectionable, because the human embryo must be destroyed
in order to harvest its stem cells.
Embryonic Germ Cells - morally objectionable when utilizing fetal tissue derived from elective
abortions, but morally acceptable when utilizing material from spontaneous abortions (miscarriages) if
the parents give informed consent.
Umbilical Cord Stem Cells - morally acceptable, since the umbilical cord is no longer required once
the delivery has been completed.
Placentally-Derived Stem Cells - morally acceptable, since the afterbirth is no longer required after the
delivery has been completed.
Adult Stem Cells - morally acceptable.

Stem Cell Banking
The cord blood of a new-borne containing stem cells- collected immediately after birth
and are preserved for future medical use.
This process involves reserving the new-born child’s umbilical cord and placenta
immediately after birth.
 The process involves collection of the blood in a collection bag and appropriately
preserved.
These cells are biologically newer and much flexible in comparison to adult stem cells,
medical fraternity uses these stem cells

Stem Cell Banking
Exceptional abilities of Umbilical Cord Stem Cells:
i) Fewer risk of complications when used in transplants
ii) Capacity to use one’s own stem cells for circumstances that lack treatment options, also
known as “autologous transplantation”
iii) Instantly available and can curtail disease progression in early treatment
Patient’s own stem cells can be used to help her/his body to prevent the future life-
threatening diseases, as no concern that her/his body will discard his own stem cells or
counter against them

Steps in cord blood banking
Cutting of Umbilical cord
Collection of cord blood
After the placenta is
delivered, the cord tissue is
then collected
Within 36 to 48 hours of collection,
the cord blood and tissue are tested,
processed
Cryogenic preservation,
at temperatures below
−170°C

Applications of Stem Cell Technology
Treatment of neural diseases such as Parkinson's disease, Huntington’s disease and
Alzheimer's disease.
Treatments for spinal cord injury, heart failure, retinal and macular degeneration, tendon
ruptures, and diabetes type 1
Stem cells could be used to repair or replace damaged neurons.
Repair of damaged organs such as the liver and pancreas.
Treatments for AIDS
Prevention and treatment of birth defects
Toxicity testing
Personalized Medicine

Stem cells are progenitor cells that are capable of self renewal
and differentiation into many different cell lineages
Stem cells have potential for treatment of many malignant and
non-malignant diseases
Third Lesson Summary

Self-healing concrete
Fourth Lesson
Topic 1
Concrete
Topic 2
systems
Topic 3
Bio mineralization
Topic 4
Microbial induced
carbonate precipitation
Topic5
Applications

Concrete
Concrete-most important building material for infrastructure
Most concrete structures are prone to cracking.
Tiny cracks on the surface of the concrete make the whole structure
vulnerable because water seeps in to degrade the concrete and corrode the
steel reinforcement,
Greatly reducing the lifespan of a structure
Any process whereby concrete recovers its performance after initial damage
is termed self-healing in concrete
Self-healing leads to a longer material lifetime, and it involves no repair and
maintenance costs.

Self-healing concrete systems
The self-healing system in concrete is principally divided into two types, autogenic and autonomic
Autogenic self-healing in concrete is an intrinsic material-healing property wherein the self-healing
process initiates from the generic materials present.
For example, cementitious materials exhibit a self-repairing ability due to the rehydration property
of unhydrated cement remaining on the crack surface.
Autonomic self-healing- Self-healing process that involves the incorporation of material
components that are not traditionally used in the concrete

systems

Bio mineralization
Process by which living organisms produce minerals.
Could be silicates in algae and diatoms, carbonates in invertebrates and calcium, phosphates and
carbonates in vertebrates.
Synthesis of minerals by prokaryotes is broadly classiﬁed into two classes:
1. Biologically controlled mineralization (BCM)
2. Biologically induced mineralization (BIM).
BCM- Minerals are directly synthesized at a speciﬁc location either within or on the cell and only
under certain conditions .
BIM- Minerals are formed extracellularly as a result of metabolic activity of the organism

BIOINDUSTRY APPLICATIONS

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