Microbial

Biotechnology
 Pasteur’s definition:
“life without air”,
anaerobe red-ox
reactions in organisms
What is
fermentation? New definition: a form
of metabolism in
which the end
products could be
further oxidized
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Fermentation
techniques
Techniques for large-scale production of microbial
products. It must both provide an optimum
environment for the microbial synthesis of the
desired product and be economically feasible on a
large scale. They can be divided into surface
(emersion) and submersion techniques. The latter
may be run in batch, fed batch, continuous reactors

In the surface techniques, the microorganisms are

cultivated on the surface of a liquid or solid
substrate. These techniques are very complicated
and rarely used in industry

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In the submersion processes, the microorganisms
grow in a liquid medium. Except in traditional beer
and wine fermentation, the medium is held in
fermenters and stirred to obtain a homogeneous
distribution of cells and medium.

Most processes are aerobic, and for these the

medium must be vigorously aerated. All important
industrial processes (production of biomass and
protein, antibiotics, enzymes and sewage
treatment) are carried out by submersion
processes.

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Some important fermentation products
Product Organism Use

Ethanol Saccharomyces Industrial

cerevisiae solvents,
beverages
Glycerol Saccharomyces Production of
cerevisiae explosives
Lactic acid Lactobacillus Food and
bulgaricus pharmaceutical
Acetone and Clostridium Solvents/fuel
butanol acetobutylicum

-amylase Bacillus Starch

subtilis hydrolysis

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General Aspects of
Fermentation Processes
Fermenter/Bioreactor

The heart of the fermentation process is the fermenter.

In general:
Stirred vessel, H/D  3
Volume 1-1000 m3 (80 % filled)
Biomass up to 100 kg dry weight/m3
Product 10 mg/l –200 g/l
Fermenter/ bioreactor Definition

•A device in which a substrate of low

value is utilized by living cells or enzymes
to generate a product of high value
 Food & drink
Fertilizer &
pesticides Enzymes
•Applications
Metal Fuel
extraction

Chemicals Waste treatment

Medicines
 Design should not Optimal mixing with
allow contaminants low uniform shear
Requirements
of a bioreactor

Gentle heat transfer

Proper feeding of substrate without
under or overdosing
Suspension of solids

Simple construction and ability to be sterilized

Simple measuring, control and regulation

Long-term stability

Scale up flexibility
Types of
bioreactor
Bioreactors can be classified according to various criteria

Configuration

Tank (height/diameter < 3)

Column (height/diameter > 3)

Hydrodynamics
Perfect mixing

Partial mixing
No mixing

Mode of operation

Continuous
Batch
Fed-batch
Type and form of biocatalyst

Free cells
Immobilized cells

Energy input and aeration

Liquid phase
Gas phase
combined
Cross section of a fermenter for
Penicillin production ( Copyright:
http://web.ukonline.co.uk/webw
ise/spinneret/microbes/penici.ht
m
)
 

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Flow sheet of a
multipurpose
fermenter and
its auxiliary
equipment
Common
bioreactors Stirred Tank Bioreactors (STB)

Most commonly used

(70%)

Simple in design & easy

to operate
Air Lift Bioreactors (ALB)

Ideal for aerobic cultures

Fluid (air/liquid) accounts

for mixing
Fluidized Bed Bioreactors (FBB)

Three phase

Volumetric productivity is
higher
 Packed Bed
Bioreactor
(PBB)

Consists of a cylindrical column

packed with inert material

Used in vinegar production,

sewage effluent treatment &
enzymatic conversion of penicillin
Stages of the process
• Stage I – Upstream processing

preparation of liquid medium, removal of

inhibitory chemicals & sterilization

Stage II – Fermentation

Conversion of substrates to desired products with

the help of microorganisms
• Stage III – Downstream processing

Separation of cells from the fermentation broth,

Isolation, purification and concentration of the
desired product
Types of media

Synthetic (chemically defined)

All the chemicals and their concentrations are
known
Product yields are low

Complex (chemically undefined)

contain undefined constituents such as beef

extract, soybean meal, molasses etc.
Product yields are high
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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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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Required value generation in fermenters as a function of size
and productivity

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