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EXAMPLES?
Wine Production
INTRODUCTION
• Fermentation can be defined as the alteration or production of
products with the help of microorganisms.
• It is process of growing micro organisms in a nutrient media by
maintaining physiochemical cond. & thereby converting feed in to
desired end product.
• Micro organisms serves as a biocatalyst in this reactions
• They are designed to produce diff. p’ceutical such as Antibiotics,
enzymes, vitamins, amino acids, insulin etc.
•Bacteria, Actinomycetes, viruses & fungi can
be used
•Industrial Biotechnology: the process by
which large quantities of cells are grown
under aerobic or anaerobic conditions.
•The industrial microorganisms are grown
under controlled conditions with an aim of
optimizing the growth of the organism for
production of a target microbial product.
•Cell growth, the structure of intracellular
enzymes, and product formation depend on
the nutritional needs of the cell (salts, oxygen)
and on the maintenance of optimum
biological
conditions
(temperature,
concentration of reactants, and pH) within
narrow limits.
FERMENTER
• Fermentation is carried out in vessels known as Fermentors
• The types of fermentor ranges from simple tank to complex
integrated system of automated control.
• contains the media to carry out fermentation, and creates
environment for fermentation at large scale.
The performance of any fermenter depends on the
following key factors:
• Agitation rate
• Oxygen transfer
• pH
• Temperature
• The design and mode of operation of a fermenter mainly
depends on the production organism, the optimal
operating condition required for target product
formation, product value and scale of production.
Pure culture:
organism, quantity, physiological state
Sterilised medium: for microorganism growth
Seed fermenter:
inoculum to initiate process
 Production fermenter:
Equipment
ii) cell separation
large model
i) drawing the culture medium
iii) collection of cell
iv) product purification v) effluent treatment.
Microbial Growth Kinetics
•Microbial Growth Kinetics describe how the
microbe grows in the fermenter.
•This information is important to determine optimal
batch times.
•The growth of microbes in a fermenter can be
broken down into four stages:
• Lag Phase
• Exponential Phase
• Stationary Phase
• Death Phase
Batch Fermentation Process
Microbial Growth Kinetics
• Lag Phase
• This is the first phase in the fermentation process
• The cells have just been injected into a new environment and they need time
to adjust accordingly
• Cell growth is minimal in this phase.
Microbial Growth Kinetics
• Exponential Phase
• The second phase in the fermentation process
• The cells have adjusted to their environment and rapid growth takes place
• Cell growth rate is highest in this phase
Microbial Growth Kinetics
• Stationary phase
• This is the third phase in the fermentation process
• The cell growth rate has leveled off and become constant
• The number of cells multiplying equals the number of cells dying.
Microbial Growth Kinetics
•Death phase
• The fourth phase in the fermentation process
• The number of cells dying is greater than the number of
cells multiplying
• The cause of the death phase is usually that the cells have
consumed most of the nutrients in the medium and there
is not enough left for sustainability
Types of Fermentation Process
• Batch Fermentation
• Continuous Fermentation
• Fed batch
BATCH FERMENTATION
• Batch reactors ,simplest type.
• Reactor is filled with medium and the fermentation is allowed.
• sterilized media components are supplied at the beginning of the
fermentation with no additional feed after inoculation.
• Once the desired amount of product is present in the fermenter the
contents are drained off and the product is extracted.
• The reactor is then cleaned, re-filled, re-inoculated and the
fermentation process starts again.
CONTINUOUS FERMENTATION
• Continuous reactors, where fresh media is continuously added and
bioreactor fluid is continuously removed.
• Sterile medium is added to the fermentation with a balancing
withdrawal of broth for product extraction.
• As a result, cells continuously receive fresh medium and products
and waste products and cells are continuously removed for
processing.
• The reactor can thus be operated for long periods of time without
having to be shut down.
• Many times more productive than batch reactors.
CONTINUOUS FERMENTATION
• does not have to be shut down as regularly
• the growth rate of the bacteria in the reactor can be more easily
controlled and optimized
• cells can also be immobilized in continuous reactors, to prevent their
removal.
Continuous culture apparatus
FED BATCH
• Fed batch reactor, most common type of reactor used in industry.
• fresh media is continuous or sometimes periodically added.
PROCESS OF FERMENTATION
• Fermentation process mainly divided into 3 steps
1)Upstream
2)Fermentation
3)Down stream
An overview of a typical industrial fermentation process and the movement of
materials through a typical fermentation plant are shown in the following figure:
Fermentation Plant
I. Upstream process
• All the operations before starting the fermenter are collectively
called upstream process.
• such as sterilization of the fermenter, preparation and sterilization of
culture medium and the preparation and growth of a suitable
inoculums of microbial strain.
Upstream Processing
Three main areas:
A) Producer microorganism
• This include processes for obtaining a suitable
• Microorganism strain improvement to increase the productivity and
yield
• Maintenance of strain purity
• Preparation of suitable inocullum
B ) Fermentation media
C) Fermentation Process
Inoculums
•Incoculum is the substance/ cell culture that is
introduced to the medium. The cell then grow in the
medium, conducting metabolisms.
•Inoculum is prepared for the inoculation before the
fermentation starts.
•It needs to be optimized for better performance:
• Adaptation in the medium
• Mutation (DNA recombinant, radiation, chemical
addition)
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Medium composition
Fermentation medium consists of:
• Macronutrients (C, H, N, S, P, Mg sources  water,
sugars, lipid, amino acids, salt minerals)
• Micronutrients (trace elements/ metals, vitamins)
• Additional factors: growth factors, attachment
proteins, transport proteins, etc)
For aerobic culture, oxygen is sparged
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Fermenter
Downstream Processing
• The processes that follows fermentation:
• A) Cell harvesting
• B) Cell disruption
• C) Product purification from cell extracts or the growth medium
III. Downstream process
• All the operations after the fermentation are known as downstream
process.
• It includes the purification of fermented products from fermentation
broth.
• Methods commonly used are distillation, centrifugation, filtration,
and solvent extraction.
FERMENTER
FERMENTATION
• The function of the fermenter or bioreactor is to provide a suitable
environment in which an organism can efficiently produce a target
product—the target product might be
• · Cell biomass
• · Metabolite
• · Bioconversion Product
• The sizes of the bioreactor can vary over several orders of
magnitudes.
• The microbial cell culture (few mm3), shake flask ( 100 -1000
ml), laboratory fermenter ( 1 – 50 L), pilot scale (0.3 – 10 m3)
to plant scale ( 2 – 500 m3) are all examples of bioreactors.
• Large volume and low value products like alcoholic
beverages need simple fermenters and do not need
aseptic condition.
• High value and low volume products require more
elaborate system of operation and aseptic condition.
• The Designing of a Bioreactor also has to take into
considerations the Unique Aspects of Biological
Processes:
A) The concentrations of starting materials
(substrates) and products in the reaction mixture are
frequently low; both the substrates and the products
may inhibit the process.
• Cell growth, the structure of intracellular enzymes, and
product formation depend on the nutritional needs of the
cell (salts, oxygen) and on the maintenance of optimum
biological conditions (temperature, concentration of
reactants, and pH) within narrow limits.
B) Certain substances, inhibitors, effectors, precursors,
metabolic products influence the rate and the mechanism of
the reactions and intracellular regulation.
C) Microorganisms can metabolize unconventional or even
contaminated raw materials (cellulose, molasses, mineral oil,
starch, ores, wastewater, exhaust air, biogenic waste), a
process which is frequently carried out in highly viscous
media.
D) In contrast to isolated enzymes or chemical catalysts,
mo’s adapt the structure and activity of their enzymes to
the process conditions, whereby selectivity and
productivity can change.
• Mutations of the microorganisms can occur under sub
optimal biological conditions.
E ) Microorganisms are frequently sensitive to strong
shear stress and to thermal and chemical influences.
REQUIREMENTS OF FERMENTERS
• There is no universal bioreactor.
• The general requirements of the bioreactor are as follows:
A) The design and construction of bioreactors must keep sterility
from the start point to end of the process.
B) Optimal mixing with low, uniform shear.
C) Adequate mass transfer, oxygen.
D) Clearly defined flow conditions.
E) Feeding substrate with prevention of under or overdosing.
F) Suspension of solids.
G) Gentle heat transfer.
H) Compliance with design requirements such as: ability to be
sterilized; simple construction; simple measuring, control,
regulating techniques; scale-up; flexibility; long term stability;
compatibility with up- downstream processes; antifoaming
measures.
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