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Food
Biotechnology

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Definition:
 Food biotechnology is the use of technology
to modify the genes of our food sources. Our
food sources are animals, plants, and
microorganisms. With food biotechnology, we
create new species of animals and plants, for
example, specifically animals and plants that
we eat. These new species have desired
nutritional, production, and marketing
properties.
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Introduction:
Food Biotechnology is the application of technology
to modify the genes of plants, animals, and
microorganisms to improve the quality, productivity,
and nutritional content of food. It combines
knowledge from biology, chemistry, and engineering
to address challenges in food production, safety, and
sustainability. Some common examples include
genetically modified organisms (GMOs), biofortified
crops, and microbial fermentation processes .

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Introduction:
Key objectives of food biotechnology
include:
 Improving crop yields by making plants

more resistant to pests, diseases, and

environmental conditions.
 Enhancing nutritional content by adding

vitamins, minerals, or other nutrients to

food crops (e.g., Golden Rice enriched with
Vitamin A).
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 Reducing food waste by extending shelf life
or improving resistance to spoilage.
 Increasing food safety by using

biotechnology to detect pathogens and

toxins in food products.
 Sustainable agriculture through reduced use

of chemical pesticides and fertilizers.

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History:
 The term “ biotechnology” was coined by a Hungarian
engineer Karl Ereky, in 1919, to refer to the science and
methods that permit products to be produced from raw
materials with the aid of living organisms.
 The biotechnology came into being centuries ago when
plants and animals began to be selectively bred and
microorganisms were used to make beer, wine, cheese,
and bread. However, the field gradually evolved, and
presently it is the use or manipulation of living
organisms to produce beneficiary substances which may
have medical, agricultural, and/or industrial utilization

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History:
 The foundation of biotechnology was laid
down after the discovery of structure of DNA
in the early 1950s. The hereditary material is
deoxyribonucleic acid (DNA) which contains
all the information that dictates each and every
step of an individual’s life. The DNA consists
of deoxyribose sugar, phosphate, and four
nitrogenous bases (adenine, guanine, cytosine,
and thymine).

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Traditional Food Fermentation:
Traditional food fermentation is the process of creating
food or changing the properties of food using microbes,
such as bacteria, yeast and fungi to convert
carbohydrate in food in to alcohol or acids. This
process has been used for century to preserve and
enhance the flavor of food.
Fermentation can also be used to produce probiotics,
which are beneficial bacteria that can help in digestion
and overall health.

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Fermentation/Industrial Organism:
 Industrial fermentation is the intentional use of fermentation
in manufacturing processes.
 In addition to the mass production of fermented foods and
drinks, industrial fermentation has widespread applications in
chemical industry.
 industrial fermentation may also involve cell culture from
plants and animals, such as CHO cells and insect cells.
Special considerations are required for the specific organisms
used in the fermentation, such as the dissolved oxygen level,
nutrient levels, and temperature. The rate of fermentation
depends on the concentration of microorganisms, cells,
cellular components.

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Fermentation Raw Materials:
 Fermentation requires specific raw materials depending on
what you're fermenting. Generally, you'll need a source of
carbohydrates (like sugar or starch), water, and
microorganisms (such as yeast or bacteria) to convert the
sugars into alcohol, acids, or other desired compounds. For
example, in beer production, malted barley provides the
carbohydrates, water is the solvent, and yeast is the
microorganism responsible for fermentation. For yogurt,
milk supplies the sugars, water, and proteins, while specific
bacteria cultures initiate the fermentation process. If you
have a specific fermentation process in mind, feel free to ask
for more detailed information.

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Submerged Fermentation:
 Submerged fermentation is a process in biotechnology
where microorganisms, such as bacteria, fungi, or
yeast, are grown in a liquid medium. This medium
typically contains nutrients like sugars, minerals, and
other necessary components for the microorganisms to
grow and produce desired substances like enzymes,
antibiotics, or biofuels. The microorganisms are
submerged in the liquid and allowed to grow and
metabolize under controlled conditions. This method is
widely used in industries like pharmaceuticals, food
and beverage, and biofuel production.

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Batch, Fed Batch & Continuous
Fermentation:
 Batch, fed-batch, and continuous fermentation are
three distinct methods used in biotechnology for
growing microorganisms and producing various
products.
 Each of these methods has its advantages and is

chosen based on the specific requirements of the

process, including the type of microorganism used,
the product being produced, and economic
considerations. Brief overview of each is given in the
following slides.

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Batch, Fed Batch & Continuous
Fermentation:
 Batch Fermentation:
In a batch fermentation, all
the necessary nutrients and microorganisms are added
to a vessel at the beginning of the process. The culture
is allowed to grow under controlled conditions until the
desired product is produced or the growth levels off
due to reduction of nutrients or accumulation of waste
products. At the end of the process, the entire contents
of the vessel are harvested, and the vessel is cleaned for
the next batch.

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Batch, Fed Batch & Continuous
Fermentation:
 Fed-Batch Fermentation:
Fed-batch fermentation
is an extension of batch fermentation. In this method,
nutrients are added incrementally over time, rather than
all at once. This allows for a longer fermentation
period, as nutrients can be supplied to the culture as
needed to sustain growth and production. Fed-batch
processes are often used when the accumulation of
certain byproducts or the depletion of nutrients limits
the growth in a batch fermentation.

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Batch, Fed Batch & Continuous
Fermentation:
 Continuous Fermentation:
In continuous fermentation,
fresh nutrients are continuously added to the system,
and an equal volume of culture is simultaneously
removed. This maintains a steady-state condition,
allowing for continuous growth and production.
Continuous fermentation is especially useful for
processes that require a consistent supply of the
product, and it's also efficient in terms of space and
resource utilization.

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Fermenter Design & Construction:
 Fermenter, also known as a bioreactor, is a
specialized vessel used in biotechnology for the
cultivation of microorganisms. Its design and
construction are crucial for maintaining optimal
conditions for microbial growth and product
formation. Here are some key aspects of fermenter
design and construction:
 Material: Fermenters are typically made from
stainless steel or other corrosion-resistant materials.
This ensures that they can withstand the harsh
conditions inside, including acidic or alkaline
environments.
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Fermenter Design & Construction:
 Vessel Shape: The shape of a fermenter can vary, but
common designs include cylindrical and conical
vessels. The choice of shape depends on factors such
as mixing requirements, heat transfer efficiency, and
ease of cleaning.
 Agitation System: Fermenters are equipped with an

agitation system to ensure uniform mixing of

nutrients, oxygen, and microorganisms. This may
include impellers, stirrers, or other types of mixing
devices.

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Fermenter Design & Construction:
 Aeration and Oxygenation: Aeration is crucial for
providing oxygen to the microorganisms. Fermenters
have spargers or diffusers that introduce air or
oxygen into the culture medium.
 Temperature Control: Temperature control systems,

including heating and cooling jackets, are

incorporated to maintain the optimal temperature for
microbial growth. This prevents temperature
fluctuations that could adversely affect the process.

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Fermenter Design & Construction:
 pH and pH Control: Fermenters are equipped with
pH sensors and controllers to monitor and adjust the
pH of the culture medium. This is important as pH
levels can impact the growth and metabolism of
microorganisms.
 Nutrient Addition: Ports or systems are included for

the addition of nutrients, including sugars, salts, and

other essential components required for microbial
growth.

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Fermenter Design & Construction:
 Sampling Ports: Sampling ports allow for periodic
sampling of the culture to monitor parameters like
cell density, pH, and product concentration.
 Sterilization and Sterile Operations: Fermenters

need to be thoroughly sterilized before use to prevent

contamination. They have systems for steam
sterilization, and some components may be
autoclavable.

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Fermenter Design & Construction:
 Monitoring and Control Systems: Fermenters are
equipped with sensors and control systems for
monitoring and regulating various parameters, such
as temperature, pH, agitation speed, and aeration rate.

 Safety Features: Safety measures, including pressure

relief valves and alarms for abnormal conditions, are
integrated to ensure the well-being of operators and
the integrity of the process.

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Fermenter Design & Construction:
 Cleaning and Maintenance: Fermenters are
designed to be easily cleaned and maintained to
prevent contamination and ensure the longevity of the
equipment.
 The design and construction of a fermenter are made-

to-order to the specific requirements of the

bioprocess it will be used for, taking into
consideration factors like the type of microorganism,
the desired product, and the scale of production.

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Types of Fermenter:
 There are several types of fermenters used in
biotechnology and industrial processes. They vary in
design, operating principles, and scale. Here are some
common types:
 Batch Fermenter:

A batch fermenter is a closed vessel

where all the necessary nutrients, microorganisms, and
other components are added at the beginning of the process.
The fermentation proceeds until the desired product is
produced or the growth levels off due to nutrient depletion
or waste accumulation.

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Types of Fermenter:
 Fed-Batch Fermenter:
This type of fermenter is an extension
of the batch fermenter. Nutrients are added incrementally over
time to sustain growth and production, allowing for a longer
fermentation period.
 Continuous Stirred-Tank Reactor (CSTR):

CSTRs are
characterized by continuous input and output of materials. Fresh
nutrients are continuously added, and an equal volume of culture
is simultaneously removed to maintain steady-state conditions.
They are particularly suitable for processes requiring a
continuous supply of the product.

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Types of Fermenter:
 Packed-Bed Fermenter:
Packed-bed fermenters use a solid
support matrix, such as beads or granules, to which
microorganisms attach and grow. The substrate is
continuously passed through the bed, providing nutrients to
the microorganisms.
 Fluidized-Bed Fermenter:

In a fluidized-bed fermenter, the

solid support material is suspended in the culture medium,
creating a fluidized state. This allows for better mixing and
mass transfer, promoting high-density cell growth.

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Types of Fermenter:
 Air-Lift Fermenter:
Air-lift fermenters use a combination
of airlift pumps and risers to circulate the culture medium and
promote mixing. This design reduces the need for mechanical
agitation and can be efficient for certain types of cultures.

 Rotary Drum Fermenter:

This type of fermenter consists
of a rotating drum filled with a substrate. The microorganisms
grow on the surface of the substrate as it moves through the
drum. It's often used for solid-state fermentation processes.

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Types of Fermenter:
 Photo-Bioreactor:
Photo-bioreactors are specialized
fermenters used for growing photosynthetic
microorganisms like algae. They are equipped with
light sources to provide the necessary energy for
photosynthesis.
 Airlift Photo bioreactor:

Airlift photo bioreactors

combine features of both airlift fermenters and photo-
bioreactors. They use airlift technology for mixing and
are designed for the cultivation of photosynthetic
microorganisms.
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Types of Fermenter:
 Hybrid Fermenters:
Hybrid fermenters combine
different principles from various types of fermenters to
optimize conditions for specific processes or products.
 The choice of fermenter type depends on factors such

as the type of microorganism, the nature of the

product, the required fermentation conditions, and the
scale of production. Each type has its advantages and
is suited to specific applications.

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Control of Physical & Chemical
Parameters in Fermenters:
 Controlling physical and chemical parameters in a
fermenter is crucial for successful fermentation
processes. Here are some key parameters to monitor
and control:
 Temperature: Maintain a consistent temperature to

ensure optimal growth and activity of the

microorganisms involved in fermentation.
 pH Level: Monitor and adjust the pH to create the

ideal environment for the specific microorganisms.

Different organisms thrive at different pH levels.

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Control of Physical & Chemical
Parameters in Fermenters:
 Agitation and Aeration: Proper mixing (agitation) and
providing adequate oxygen (aeration) are essential for
uniform distribution of nutrients and maintaining suitable
conditions.
 Nutrient Levels: Ensure that essential nutrients like sugars,
nitrogen sources, and minerals are provided in appropriate
quantities to support microbial growth and metabolism.
 Oxygen Levels: Depending on the type of fermentation
(aerobic or anaerobic), control the oxygen levels
accordingly. Some organisms require oxygen, while others
do not.

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Control of Physical & Chemical
Parameters in Fermenters:
 Foaming: Use antifoaming agents or adjust agitation to
prevent excessive foaming, which can hinder efficient
mixing and oxygen transfer.
 Pressure: Maintain a consistent pressure within the

fermenter to ensure proper gas exchange and prevent

any potential issues related to vessel integrity.
 Dissolved Gas Levels: Monitor the concentration of

gases like oxygen and carbon dioxide in the culture

medium. This is particularly important for aerobic
fermentations.

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Control of Physical & Chemical
Parameters in Fermenters:
 Residence Time: Control the time microorganisms
spend in the fermenter. This affects productivity and the
overall efficiency of the process.
 Product Concentrations: Regularly measure the
concentrations of desired products (e.g., ethanol,
organic acids) to track the progress of the fermentation.
 Sterilization and Sanitation: Ensure proper
sterilization(cleaning) of the fermenter and its
components before vaccination to prevent
contamination.

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Control of Physical & Chemical
Parameters in Fermenters
 Sampling and Analysis: Take samples at regular
intervals to analyze the culture for microbial
population, nutrient levels, pH, and other relevant
parameters.
 Remember, precise control of these parameters

depends on the specific type of fermentation and the

microorganisms involved. Regular monitoring and
adjustments are essential to optimize the fermentation
process.

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Down Stream Processing; Product Recovery,
Purification & Standardization:
 Certainly Downstream processing encompasses several crucial
steps, including product recovery, purification, and
standardization:
 Product Recovery: Product Recovery involves the following Steps:
 Cell Separation: This initial step involves separating cells from
the culture medium, typically using techniques like centrifugation
or filtration.
 Cell Disruption: If th e desired product is inside the cells, they
need to be disrupted to release it. This can be achieved through
methods like homogenization, sonication, or enzymatic digestion.
 Clarification: This step removes any remaining solid particles,
cell debris, and other impurities, usually through techniques like
centrifugation, filtration, or sedimentation.
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Down Stream Processing; Product Recovery,
Purification & Standardization:
 Purification: Purification involves the following steps:
 Chromatography: This is a powerful technique used to
separate molecules based on their affinity for a stationary
phase. Techniques like affinity, ion exchange, size exclusion,
and other forms of chromatography are employed to purify the
target product.
 Precipitation: By adjusting factors like pH, temperature, or
the addition of specific reagents, proteins and other
biomolecules can be made to precipitate out of solution,
allowing for their isolation.
 Filtration: Additional filtration steps may be used to further
refine the product and remove smaller impurities.

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Down Stream Processing; Product Recovery,
Purification & Standardization:
 Concentration: After purification, the product may be
concentrated to increase its potency and reduce its volume for
storage and transportation. Techniques like ultrafiltration and
diafiltration are often used.
 Standardization: Standardization includes the following
steps
 Analytical Techniques: Various analytical methods are employed
to assess the quality and quantity of the product. This can include
techniques like high-performance liquid chromatography (HPLC),
mass spectrometry, and spectroscopy.
 Quality Control: Strict quality control measures are applied to
ensure the final product meets defined specifications for purity,
potency, and safety.
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Down Stream Processing; Product Recovery,
Purification & Standardization:
 Formulation and Final Processing: Depending on the
application, the purified product may undergo further
processing steps to formulate it into its final dosage form.
This can include steps like buffer exchange, lyophilization
(freeze-drying), and sterile filtration.
 :Documentation and Regulatory Compliance: Thorough
documentation of all steps, including standard operating
procedures (SOPs) and batch records, is essential for
regulatory compliance.
 Stability Testing: The final standardized product undergoes
stability testing to determine its shelf life and storage
conditions.

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Down Stream Processing; Product Recovery,
Purification & Standardization:
 Packaging and Labeling: The final product is
packaged and labeled according to regulatory
guidelines and industry standards.
 These steps are critical in ensuring that the final

product is of high quality, safe for use, and meets the

required specifications for its intended application.
Keep in mind that the specific techniques and
technologies used can vary depending on the type of
product and the manufacturing process.

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Solid Substrate Fermentation:
 Solid substrate fermentation (SSF) is a bioprocessing
technique used for the production of various biochemicals,
enzymes, and microbial biomass. Unlike submerged
fermentation, which takes place in a liquid medium, SSF
involves the cultivation of microorganisms on a solid
substrate. Here's a basic overview of the process:
 Substrate Selection: A solid substrate, typically
composed of natural materials like grains, agricultural
residues, or synthetic materials, is chosen. It provides both
a physical support for microbial growth and a source of
nutrients.

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Solid Substrate Fermentation:
 Moisture Adjustment: The substrate is moistened to
a level that supports microbial growth but doesn't
cause excessive water accumulation. This is crucial
for the metabolic activities of the microorganisms.
 Inoculation/Vaccination: The microorganisms
(usually fungi or bacteria) are inoculated onto the
solid substrate. These microorganisms can be native
to the substrate or introduced specifically for the
fermentation process.

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Solid Substrate Fermentation:
 Incubation/Growth: The inoculated/vaccinated substrate is
placed in a controlled environment with appropriate temperature,
humidity, and aeration conditions. This allows the microorganisms
to grow, metabolize, and produce the desired product.
 Fermentation Progress: As the microorganisms grow, they
enzymatically convert the substrate into desired products. This can
include enzymes, organic acids, bioactive compounds, or other
metabolites.
 Harvesting: Once the fermentation process is complete, the
product is typically harvested by separating it from the residual
substrate. This can involve physical methods like grinding,
sieving, or extraction.

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Solid Substrate Fermentation:
 Solid Substrate Fermentation (SSF) offers several advantages:
 Low Water Requirement: SSF typically uses less water
compared to submerged fermentation, making it more
environmentally sustainable, especially in regions with water
scarcity.
 Reduced Contamination Risk: The solid substrate provides
a natural barrier against contaminants, reducing the risk of
unwanted microbial growth.
 Enhanced Product Concentration: The solid nature of the
substrate allows for higher product concentrations, which can
simplify downstream processing.

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Solid Substrate Fermentation:
 SSF (Solid Substrate Formentation) is used in various
industries, including food and beverage,
pharmaceuticals, biofuels, and enzyme production. It's
particularly well-suited for the production of certain
enzymes, organic acids, and secondary metabolites.
 Keep in mind that the specific parameters and

techniques used in SSF (Solid Substrate Formentation)

can vary depending on the microorganism and product
being produced, as well as the characteristics of the
chosen substrate.

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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 The production of organic acids, enzymes, amino acids,
single-cell protein, carotenoids, and fermented food
products involves various bioprocessing techniques.
Here's a brief overview of how each of these products is
typically produced:
 Organic Acids Submerged Fermentation:
Microorganisms like bacteria or fungi are cultivated in a
liquid medium containing a carbon source. They
metabolize the carbon source and produce organic acids
as metabolic byproducts. Common organic acids
produced include citric acid, lactic acid, and acetic acid.

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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 Production of Citric Acid is an industrial process that uses
raw materials like substrates, citric acid growth-promoting
microorganisms and enzymes etc. for the commercial
production of citric acid. Generally, the commercial
production of citric acid works out best by employing the
fermentation method.
 Globally, there is around 7, 36,000 tonnes/year production of
citric acid. The citric acid is commercially produced because
of its broad applicability in human consumption (increased by
4% each year) and its high demand in food, pharmaceuticals,
and other industries like cosmetics, toiletries etc

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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 Itaconic Acid is traditionally produced by
fermentationof carbohydrates using vaious varities of
the genus Aspergillus, especially Aspergillus terreus
and Aspergillus itaconicus. Itaconic acid is produced
from a citric acid cycle intermediate, Cis-aconitate,
which is formed from citric acid. It can be produced
from sugars such as glucose by breaking down
glucose into pyruvate by glycolysis, which is further
converted into acetyl-CoA with the release of
CO2power2.

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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 Enzymes Fermentation or Solid Substrate
Fermentation (SSF): Enzyme-producing
microorganisms are cultivated in a suitable medium.
After growth, the enzymes are harvested from the
culture broth or solid substrate. Downstream
processing may involve purification and concentration.
 Amino Acids, Submerged Fermentation: Specific

amino acid-producing microorganisms are grown in a

liquid medium containing precursors necessary for
amino acid synthesis. The amino acids are then
extracted and purified.
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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 Single-Cell Protein (SCP), Solid Substrate
Fermentation (SSF) or Submerged Fermentation:
Microorganisms with high protein content are
cultivated on a suitable substrate (solid or liquid). The
biomass is harvested and processed into a protein-rich
product suitable for human or animal consumption.
 Carotenoids, Submerged Fermentation: Certain

microorganisms, like algae, bacteria, or fungi, are

grown in a liquid medium under specific conditions to
stimulate carotenoid production. The carotenoids are
then extracted and purified.
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Production of Organic Acids (citric acid & itaconic acid),
Enzymes, Amino Acids, Single Cell Protein, Carotenoids &
Fermented Food Products:
 Fermented Food Products, Fermentation of Raw Materials: This
process involves the conversion of raw food materials by
microorganisms to produce a range of food products like bread,
cheese, yogurt, kimchi, and more.
 Lactic Acid Fermentation: Lactic acid bacteria are commonly used
in the fermentation of dairy products, vegetables, and certain meats.
 Alcoholic Fermentation: Yeasts are used to convert sugars into
alcohol, which is employed in brewing and winemaking. Remember,
the specific microorganisms, fermentation conditions, and
downstream processing steps will vary depending on the product
and industry. Quality control measures, regulatory compliance, and
product standardization are also crucial aspects of the production
process.

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GMO (Genetic Modification
Techniques)in Food Biotechnology:
 Genetic modification techniques in food
biotechnology involve altering the genetic material of
organisms to impart specific traits or characteristics.
One common method is recombinant DNA technology.
This involves isolating and modifying genes from one
organism (like a bacteria or plant) and inserting them
into the genome of another organism (like a crop
plant). This allows scientists to introduce traits like pest
resistance, drought tolerance, or improved nutritional
content.

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GMO (Genetic Modification
Techniques)in Food Biotechnology:
 Another technique is CRISPR-Cas9. It's a precise and
versatile tool that can edit specific genes within an
organism's DNA. In agriculture, it's used to create crops
with desirable traits, such as disease resistance or enhanced
yield.These techniques have been instrumental in
developing genetically modified organisms (GMOs) like
insect-resistant Bt cotton, herbicide-tolerant soybeans, and
nutritionally enriched crops. However, GMOs are a subject
of debate due to concerns about safety, ethics, and
environmental impact. It's crucial to carefully assess and
regulate these technologies to ensure their responsible and
beneficial use in food production.

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Legal and Social Aspects of food
Biotechnology
 Food biotechnology involves the use of
biological processes, organisms, or systems to
develop or modify food products. This field
encompasses various aspects, including
genetic engineering, fermentation, and other
techniques to improve food production,
quality, and safety. Here are some key legal
and social aspects associated with food
biotechnology.

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Legal and Social Aspects of food
Biotechnology
 Legal Aspects:

 Regulation and Oversight: Governments and

regulatory agencies establish frameworks to ensure the
safety and quality of biotechnologically modified
foods. This involves evaluating new technologies,
conducting risk assessments, and establishing
guidelines for labeling and safety standards.

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Legal and Social Aspects of food
Biotechnology
 Labeling and Transparency: Regulations often
mandate the labeling of biotech-derived foods to
inform consumers about their content. This is aimed at
providing transparency and allowing individuals to
make informed choices.
 Intellectual Property and Patents: Biotech
companies often seek patents for their innovations,
which can lead to debates about ownership, access, and
control over genetic resources. This can have
implications for smaller agricultural communities and
biodiversity.
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Legal and Social Aspects of food
Biotechnology
 Trade Agreements and International Standards:
Agreements and standards at the international level can
impact the trade of biotech foods. Some countries may
have different regulatory approaches, leading to
potential trade disputes.
 Liability and Risk Management: Legal frameworks

need to address issues related to potential harm caused

by biotech foods, including allergenicity or unintended
environmental consequences.

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Legal and Social Aspects of food
Biotechnology
 Social Aspects:
 Consumer Perception and Acceptance: Public
perception of biotech foods can vary widely. Factors like
trust in regulatory bodies, information accessibility, and
cultural attitudes towards technology can influence
consumer acceptance.
 Ethical Concerns: Debates arise regarding the ethics of
modifying organisms for food production. This includes
considerations of animal welfare, environmental impact,
and potential unintended consequences.

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Legal and Social Aspects of food
Biotechnology
 Food Security and Access: Biotechnology can play a
role in addressing global food security challenges by
enhancing crop yields, nutritional content, and
resistance to pests or diseases. However, equitable
access to these technologies is a critical concern.
 Environmental Impact: The use of biotechnology in

agriculture can have environmental consequences. This

includes potential effects on biodiversity, soil health,
and the development of resistant pests.

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Legal and Social Aspects of food
Biotechnology
 Cultural and Socioeconomic Impacts: Biotech
interventions can have varying effects on different
communities. For example, small-scale farmers may face
challenges in adopting expensive biotech practices, while
larger agribusinesses may benefit.
 Balancing these legal and social aspects is crucial for the
responsible development and deployment of food
biotechnology. It requires collaboration between
stakeholders, including governments, scientists, industry,
and civil society, to ensure that benefits are maximized
while minimizing potential risks and negative impacts.

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PRACTICAL NO:01
 Lab Scale Production of Organic Acids i.e, Citric Acid etc:
 Producing organic acids like citric acid on a lab scale requires
specific apparatus and a well-defined procedure. Here's an outline:
 Apparatus:
 Fermentation Vessel: A vessel capable of holding the
fermentation medium and providing conditions (temperature, pH,
aeration) suitable for microbial growth and acid production.
 Agitator or Stirrer: Ensures uniform mixing of the fermentation
medium, promoting optimal conditions for the microorganism.
 pH Meter: Used to monitor and control the pH level throughout
the fermentation process.

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PRACTICAL NO:01
 Temperature Control System: Maintains a constant
temperature suitable for the chosen microorganism's
growth and acid production.
 Aeration System: Supplies oxygen to the
microorganisms, crucial for their metabolic processes.
 Inoculation Equipment: Such as a pipette or loop for
introducing the selected microorganism into the
fermentation medium.
 Monitoring Equipment: Instruments for measuring
parameters like temperature, pH, dissolved oxygen, and
sugar concentration.

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PRACTICAL NO:01
 Filtration or Separation Apparatus: Used to separate the
microorganisms from the fermentation broth after
fermentation.
 Extraction and Purification Equipment: Depending
on the specific procedure, this may include filtration,
centrifugation, and chemical agents for purification.
 Crystallization Setup: Equipment for inducing
crystallization of the organic acid from the extracted
solution.
 Drying Equipment: Used to remove moisture from the

crystallized organic acid.

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PRACTICAL NO:01
 Procedure:
 Medium Preparation: Prepare a growth medium containing
a carbon source (such as glucose) and other nutrients
(nitrogen, minerals, etc.) needed for microbial growth and
acid production.
 Sterilization: Sterilize the growth medium to eliminate any
contaminants that could interfere with the fermentation
process.
 Inoculation: Inoculate the sterilized medium with the
selected microorganism. This introduces the microorganisms
into the growth medium to start the fermentation process.

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PRACTICAL NO:01
 Fermentation: Maintain optimal conditions (temperature, pH,
aeration, agitation) for microbial growth and acid production.
Monitor parameters regularly.
 Harvesting: When the fermentation reaches its peak, separate
the microorganisms from the liquid portion (fermentation
broth).
 Extraction: Extract the organic acid from the fermentation
broth using appropriate techniques. This could involve filtration,
centrifugation, or chemical precipitation.
 Purification: Further purify the extracted organic acid if
necessary, using methods like filtration, chromatography, or
crystallization.

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PRACTICAL NO:01
 Concentration and Crystallization: Concentrate the
extracted organic acid and induce crystallization to
obtain a purer form.
 Drying: Dry the crystals to remove any remaining

moisture.
 Packaging: Package the citric acid in suitable

containers for storage and distribution.

 Always follow lab safety protocols and use appropriate

personal protective equipment when working with

microorganisms and chemicals.

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PRACTICAL NO:02
 Microbal Enzymes: Amylases and Proteases
 Apparatus and procedure for conducting experiments with
microbial enzymes like amylase and protease:
 Apparatus:
 Test Tubes: These are small glass or plastic tubes used to hold and
mix the enzyme solution with the substrate.
 Pipettes: These are used to measure and transfer precise volumes
of enzyme solutions and substrates.
 Water Bath: This is a controlled-temperature bath used to
maintain a constant temperature for the reaction.
 Stopwatch or Timer: This is used to time the duration of the
reaction.

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PRACTICAL NO:02
 pH Meter: Used to monitor and adjust the pH level of the
reaction mixture
 Thermometer: To ensure the temperature is accurately
controlled.
 Spectrophotometer (for protease experiments): This
instrument measures the absorption of light by a solution at
different wavelengths.
 Substrates: For amylase, this could be starch; for protease, it
could be a protein like casein.
 Buffers: To maintain a stable pH level.

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PRACTICAL NO:02
 Procedure for Amylase Experiment:
 Prepare Enzyme Solution: Extract or obtain a sample of
amylase enzyme. Create a known concentration by diluting it
with an appropriate buffer solution.
 Prepare Substrate Solution: Prepare a starch solution of
known concentration.
 Mix Enzyme and Substrate: In a test tube, combine the
amylase solution and the starch solution in known
proportions.
 Incubate: Place the test tube in a water bath set at a specific
temperature (usually around 37°C, which is body temperature)
and allow the reaction to proceed for a set period of time.

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PRACTICAL NO:02
 Stop the Reaction: This can be done by heating the mixture to
denature the enzyme or by adding a chemical that deactivates the
enzyme.
 Test for Reaction Completion: This may involve using an iodine
solution which changes color in the presence of starch. A change
from blue-black to brown indicates the absence of starch.
 Procedure for Protease Experiment:
 Prepare Enzyme Solution: Extract or obtain a sample of protease
enzyme. Create a known concentration by diluting it with an
appropriate buffer solution.
 Prepare Substrate Solution: Prepare a protein solution (e.g.,
casein) of known concentration.

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PRACTICAL NO:02
 Mix Enzyme and Substrate: In a test tube, combine the
protease solution and the protein solution in known
proportions.
 Incubate: Place the test tube in a water bath set at a

specific temperature (usually around 37°C) and allow

the reaction to proceed for a set period of time.
 Stop the Reaction: This can be done by adjusting the

pH or by adding a chemical that deactivates the

enzyme.

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PRACTICAL NO:02
 Measure Absorbance: Using a spectrophotometer,
measure the absorbance of the solution at a specific
wavelength. The change in absorbance indicates the
progress of the reaction. Remember, the exact
concentrations, temperatures, and times will vary
depending on the specific experiment and the
characteristics of the enzymes and substrates you're
working with. Always follow established protocols
and safety guidelines when conducting experiments.

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PRACTICAL NO:03
 Single Cell Proteins and Fermented Food
Products through Biotechnological Applications:
 1. Single Cell Proteins through Biotechnological Applications:
 Producing single-cell proteins (SCP) through biotechnological
applications involves specific equipment and procedures.
 Apparatus:
 Fermenter or Bioreactor: This is a vessel where
microorganisms are cultured under controlled conditions. It has
features like temperature control, pH monitoring, agitation, and
aeration systems.

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PRACTICAL NO:03
 Sterilization Equipment: Autoclaves or other sterilization
devices are used to ensure that all equipment and culture
media are free from contaminants before starting the
fermentation process.
 Nutrient Feed System: This includes tanks and pumps for
providing the microorganisms with necessary nutrients like
sugars, nitrogen sources, minerals, etc.
 Monitoring and Control Systems: These include sensors for
measuring parameters like pH, temperature, dissolved oxygen,
and agitation speed. They help maintain optimal conditions for
microbial growth.

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PRACTICAL NO:03
 Harvesting Equipment: This can range from centrifuges to separate the
microbial biomass from the fermentation broth, to filtration systems.
 Drying or Processing Equipment: Depending on the desired form of
the final product, equipment for drying or further processing may be
required.
 Procedure:
 Microbial Selection: Choose a suitable microorganism (bacteria, yeast,
or fungi) based on its ability to efficiently convert raw materials into
protein-rich biomass.
 Media Preparation: Prepare a nutrient-rich growth medium containing
carbohydrates, nitrogen sources, minerals, and other necessary
components. Sterilize the media to eliminate any potential contaminants.

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PRACTICAL NO:03
 Inoculation: Introduce the selected microorganism into the
bioreactor containing the sterilized growth medium.
 Fermentation: Control the conditions within the bioreactor to
facilitate the growth and multiplication of the microorganism.
This includes regulating factors like temperature, pH, agitation
speed, and aeration rate.
 Nutrient Addition: Monitor nutrient levels and add additional
nutrients as needed to sustain microbial growth.
 Biomass Harvesting: Once the fermentation process is
complete, harvest the microbial biomass. This can be done
using centrifugation, filtration, or other suitable methods.

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PRACTICAL NO:03
 Drying or Further Processing: Depending on the intended use, the
harvested biomass may undergo additional processing steps like
drying to obtain a stable and usable form.
 Quality Control: Perform tests to ensure the final product meets
quality standards, including protein content and absence of
contaminants.
 Packaging and Storage: Package the SCP in appropriate containers
and store them under conditions that maintain product integrity.
 By following these steps and using the necessary equipment, Single
Cell Proteins (SCP) can be produced through biotechnological
applications for various industrial and food-related purposes. Keep
in mind that specific details may vary depending on the
microorganism and intended application.

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PRACTICAL NO:03
 2. Fermented Food Products through Biotechnological
Applications:
 Producing fermented food products through biotechnological
applications involves specific equipment and procedures.
 Apparatus:
 Fermenter or Bioreactor: This vessel is designed to control and
optimize the conditions for microbial fermentation. It includes
features like temperature control, pH monitoring, agitation, and
aeration systems.
 Sterilization Equipment: Autoclaves or other sterilization
devices are used to ensure that all equipment and culture media are
free from contaminants before starting the fermentation process.

77
PRACTICAL NO:03
 Nutrient Feed System: This includes tanks and pumps for
providing the microorganisms with necessary nutrients like sugars,
nitrogen sources, minerals, etc.
 Monitoring and Control Systems: These include sensors for
measuring parameters like pH, temperature, dissolved oxygen, and
agitation speed. They help maintain optimal conditions for
microbial growth.
 Harvesting Equipment: Depending on the fermented product, this
could include filtration systems, separators, or other devices to
separate the final product from the fermentation broth.
 Packaging and Storage Equipment: This includes containers,
bottling machines, and sealing equipment for the final fermented
product.

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PRACTICAL NO:03
 Procedure:
 Microbial Selection: Choose a suitable microorganism
(bacteria, yeast, or fungi) based on its ability to ferment the
specific raw materials used in the production of the desired
food product.
 Media or Substrate Preparation: Prepare the raw materials
or substrates needed for the fermentation process. This could
be grains, fruits, vegetables, or other suitable sources.
 Inoculation: Introduce the selected microorganism into the
bioreactor or fermenter containing the prepared substrate.

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PRACTICAL NO:03
 Fermentation: Control the conditions within the bioreactor to
facilitate the growth and activity of the microorganism. This
includes regulating factors like temperature, pH, agitation speed,
and aeration rate.
 Nutrient Addition: Monitor nutrient levels and add additional
nutrients as needed to sustain microbial growth and activity.
 Monitoring and Sampling: Regularly monitor the progress of
fermentation by taking samples and analyzing parameters like pH,
acidity, alcohol content, etc.
 Harvesting and Processing: Once the fermentation process is
complete, harvest the fermented product. This may involve
separating the final product from the fermentation broth and, in
some cases, further processing steps.

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PRACTICAL NO:03
 Packaging and Storage: Package the fermented food product
in appropriate containers and store them under conditions that
maintain product integrity.
 Quality Control: Perform tests to ensure the final product
meets quality standards, including taste, texture, aroma, and
absence of contaminants.
 By following these steps and using the necessary equipment, a
wide variety of fermented food products can be produced
through biotechnological applications. Keep in mind that
specific details may vary depending on the type of food
product and the microorganisms involved.

81