FERMENTATION AND

BIOPROCESSING
Bhakti Bapat
INTRODUCTION:
 Branch of Microbiology which deals with the study and
use of various micro-organisms that are responsible for
the production of many products such as
 alcoholic products
 Antibiotics
 Enzymes
 vaccines

 This is done using of Genetically Modified organisms

with increased productivity. 

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APPLICATIONS OF INDUSTRIAL
MICROBIOLOGY
 Bacteria, Fungi, Yeast etc. are used commonly in the
production of various fermented products like wine, yogurt etc.
 Strains of Corynebacterium glutamicum have been used in the
production of the amino acid, L-glutamate.
 Production of food and dairy products. Cheese, yoghurt,
alcoholic beverages, coffee, tea,
 Producing vitamins,sugars, amino acids and various small
biomolecues
 Production of vaccines is another important application of
industrial microbiology.
 Antibiotics are another important products produced by using
micro- organisms. 3

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MICRO ORGANISM GROWTH IN BATCH
PROCESS

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PRIMARY METABOLITES
 Primary metabolites are involved in growth, development, and reproduction of
the organism.
 The primary metabolite is typically a key component in maintaining normal
physiological processes; thus, it is often referred to as a central metabolite.
 Primary metabolites are typically formed during the growth phase as a result of
energy metabolism, and are deemed essential for proper growth.
 Examples of primary metabolites include alcohols such as ethanol, lactic acid,
and certain amino acids.
 Alcohol is one of the most common primary metabolites used for large-scale
production. Specifically, alcohol is used for processes involving fermentation
which produce products like beer and wine.
 Additionally, primary metabolites such as amino acids– including L-glutamate
and L-lysine, which are commonly used as supplements– are isolated via the
mass production of a specific bacterial species, Corynebacteria glutamicum.
 Citric acid, produced by Aspergillus niger, is one of the most widely used
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ingredients in food production. It is commonly used in pharmaceutical and
cosmetic industries as well.
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SECONDARY METABOLITES
 Secondary metabolites are typically organic compounds
produced through the modification of primary metabolite
synthases.
 Secondary metabolites do not play a role in growth,
development, and reproduction like primary metabolites
do, and are typically formed during the end or near the
stationary phase of growth.
 These metabolites can be used in industrial microbiology
to obtain amino acids, develop vaccines and antibiotics,
and isolate chemicals necessary for organic synthesis.

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SECONDARY METABOLITES

 Many of the identified secondary metabolites have a role in ecological

function, including defense mechanism(s), by serving as antibiotics
and by producing pigments. Examples of secondary metabolites with
importance in industrial microbiology include atropine and antibiotics
such as erythromycin and bacitracin.
 Atropine, derived from various plants, is a secondary metabolite with
important use in the clinic. Atropine is used in the treatment of
bradycardia.
 Antibiotics such as erythromcyin and bacitracin are also considered to
be secondary metabolites. Erythromycin, derived
from Saccharopolyspora erythraea, is a commonly used antibiotic
with a wide antimicrobial spectrum. It is mass produced and
commonly administered orally.
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 Bacitracin, derived from organisms classified under Bacillus subtilis,
is an antibiotic commonly used a topical drug.
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FEATURE OF INDUSTRIAL
MICROORGANISM
 Genetically stable
 Efficiency
 Culture efficiency
 Product efficiency

 Simple nutritional value

 Cheap carbon and energy source

 Genetically manipulable
 Addition
 Deletion
 Insertion 
 Safety
 Easy product recovery

 Profitable by-products
 ETHANOL FERMENTATION
 Ethanol fermentation is used to produce ethanol for use in food, alcoholic
beverages, and both fuel and industry.
 The process of ethanol fermentation occurs when sugars are converted into cellular
energy. The sugars which are most often used include glucose, fructose, and sucrose.
These sugars are converted into cellular energy and produce both ethanol and
carbon dioxide as waste products.
 Yeast is the most commonly used organism to produce ethanol via the fermentation
process for beer, wine, and alcoholic drink production.
 Yeast prefer to utilize fermentation over aerobic respiration Hence, the use of yeast
on a large-scale to produce ethanol and carbon dioxide occurs in an anaerobic
environment.
 The yeast will convert the sugars produced as a result of the conversion of grain
starches to sugar by amylase. Additionally, yeast fermentation is utilized to mass
produce ethanol which is added to gasoline. The major source of sugar utilized for
ethanol production in the US is currently corn; however, crops such as sugarcane or
sugar beets can be used as well.
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OTHER RECOMBINANT PRODUCTS
 Fermentation is also utilized in the mass production of various
recombinant products. These recombinant products include numerous
pharmaceuticals such as insulin and hepatitis B vaccine.
  E. coli, which has been genetically altered to produce proinsulin, is
grown to a large amount to produce sufficient amounts in a fermentation
broth. The proinsulin is then isolated via disruption of the cell and
purified. There is further enzymatic reactions that occur to then convert
the proinsulin to crude insulin which can be further altered for use as a
medicinal compound.
 The creation of this vaccine utilizes both recombinant DNA technology
and fermentation. A gene, HBV, which is specific for hepatitis B virus, is
inserted into the genome of the organism yeast. The yeast is used to
grow the HBV gene in large amounts and then harvested and purified.
The process of fermentation is utilized to grow the yeast, thus promoting
the production of large amounts of the HBV protein which was 10
genetically added to the genome.
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BIOCHEMICAL ENGINEERING
 Biochemical engineering means, the extension of chemical
engineering principles to systems using a biological
catalyst to bring about desired chemical transformations.
 It is the branch of chemical engineering that mainly deals
with the design and construction of unit processes that
involve biological organisms as molecules.
 Applications of biochemical engineering are used
 Food
 Feed
 Pharmaceutical
 Biotechnology
 Water treatment industries 11

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UPSTREAM PROCESSING
 Upstream processing step involves
 chemical hydrolysis
 preparation of liquid medium
 separation of particulate
 air purification
 many other preparatory operations.

 Bio-reaction step
 The resulting feed is transferred to one or more Bio reaction
stages. This step is mainly consists of three operations
namely
 production of biomass
 metabolize biosynthesis
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 biotransformation.

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DOWNSTREAM PROCESSING
 Finally, the material produced in the bioreactor must be
further processed in the downstream section to convert it
into more useful form. The downstream process is mainly
consists of physical separation operations which includes:
 solid liquid separation
 Adsorption
 liquid-liquid extraction
 Distillation
 drying etc

 The raw material can be of biological or non-biological

origin. It is first converted to more suitable form for
processing.  14

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BIOREACTOR TYPES:
 Stirred Tank Bioreactors
 Bubble Column Bioreactors

 Airlift Bioreactors

 Fluidized Bed Bioreactors

 Packed Bed Bioreactors

 Trickle bed
 Photo-Bioreactors
 Rotary drum bioreactor

 Mist Bioreactor

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STIRRED TANK BIOREACTORS

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STIRRED TANK BIOREACTORS

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CONTINUOUS STIRRED TANK
BIOREACTORS

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 SPECIFICATIONS OF A BIOREACTOR AND
MODELLING
 A typical bioreactor consists of following parts:
 Agitator – used for the mixing of the contents of the reactor which keeps the
“cells” in the perfect homogenous condition for better transport of nutrients
and oxygen to the desired product(s).
 A continuous stirred tank bioreactor consists of a cylindrical vessel with motor
driven central shaft that supports one or more agitators (impellers).
 Baffle – used to break the vortex formation in the vessel, which is usually
highly undesirable as it changes the center of gravity of the system and
consumes additional power.
 Jacket – The jacket provides the annular area for circulation of constant
temperature of water which keeps the temperature of the bioreactor at a
constant value
 Sparger – In aerobic cultivation process, the purpose of the sparger is to supply
adequate oxygen to the growing cells. This enables the creation of a uniform
and homogeneous environment throughout the bioreactor. 

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STIRRED TANK REACTORS:

Advantages Disadvantages

 Efficient gas transfer to   The need for shaft seals

growing cells and bearings.
 Good mixing of the  Size limitation by motor
contents size, shaft length &
 Flexible operating weight. 
conditions
 Commercial availability
of the bioreactors
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 BUBBLE COLUMN BIOREACTOR
  In the bubble column bioreactor, the
air or gas is introduced at the base of
the column through perforated pipes
or plates, or metal micro porous
spargers.
 The flow rate of the air/gas
influences the performance factors
like O2 transfer, mixing.
 Bubble column bioreactors may be
fitted with perforated plates to
improve performance.
 The vessel used for bubble column
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bioreactors is usually cylindrical
with an aspect ratio of 4-6 L/D Bhakti Bapat
AIRLIFT COLUMN BIOREACTOR

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AIRLIFT BIOREACTORS
 Internal-loop airlift bioreactor has a single container with a
central draft tube that creates interior liquid circulation channels.
 These bioreactors are simple in design, with volume and
circulation at a fixed rate for fermentation.
 External loop airlift bioreactor possesses an external loop so that
the liquid circulates through separate independent channels.
 These reactors can be suitably modified to suit the requirements
of different fermentations.
 In general, the airlift bioreactors are more efficient than bubble
columns, particularly for more denser suspensions of
microorganisms.
 This is mainly because in these bioreactors, the mixing of the
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contents is better compared to bubble columns.

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 Two stage airlift bio reactors
 Two-stage airlift bioreactors are used for the temperature
dependent formation of products.
 Growing cells from one bioreactor (maintained at temperature
30°C) are pumped into another bioreactor (at temperature
42°C).
 There is a necessity for the two-stage airlift bioreactor, since it
is very difficult to raise the temperature quickly from 30°C to
42°C in the same vessel.
 Each one of the bioreactors is fitted with valves and they are
connected by a transfer tube and pump.
 The cells are grown in the first bioreactor and the bioprocess 25
proper takes place in the second reactor.
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APPLICATION OF AIRLIFT BIOREACTORS

 Airlift bioreactors are commonly employed for aerobic

bioprocessing technology.
 They ensure a controlled liquid flow in a recycle system by
pumping.
 Due to high efficiency, airlift bioreactors are sometimes
preferred in
 e.g.,methanol production
 waste water treatment
 single-cell protein production.

 In general, the performance of the airlift bioreactors is

dependent on the pumping (injection) of air and the liquid
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circulation.
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TOWER BIOREACTORS
 Tower fermenters are
characterized by large height
to diameter ratios, upwards of
15:1, and aeration occurs by
the introduction of pressured
gas streams at the bottom of
the tower

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AIRLIFT BIOREACTORS
Advantages Disadvantages

 Simple design  Greater air throughput

 Easier sterilization (no and higher pressures
agitator shaft parts) needed.
 Low shear effects  Extensive foaming which

 Low Energy requirement can clog the air exhaust

vs stirred tank filters and increase the
risk of contamination
 Greater heat-removal vs
stirred tank
 Very low cost  28

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FLUIDIZED BED BIOREACTOR

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 Fluidized bed bioreactor is comparable to bubble column bioreactor except the
top position is expanded to reduce the velocity of the fluid.
 The design of the fluidized bioreactors (expanded top and narrow reaction
column) is such that the solids are retained in the reactor while the liquid flows
out
 It is also necessary to ensure that the suspended solid particles are not too light or
too dense (too light ones may float whereas to dense ones may settle at the
bottom), and they are in a good suspended state.
 Recycling of the liquid is important to maintain continuous contact between the
reaction contents and biocatalysts. This enable good efficiency of bioprocessing.
 In a fluidised bed reactor, cells or enzymes are immobilised in and/or on the
surface of light particles
 These bioreactors are suitable for use to carry out reactions involving fluid
suspended biocatalysts such as immobilized enzymes, immobilized cells, and
microbial flocs. of the due to gravity. This results in good circulation 30

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APPLICATIONS OF FBB
 These bioreactors are suitable for use to carry out
reactions involving fluid suspended biocatalysts such as
immobilized enzymes, immobilized cells, and microbial
flocs.

 Both aerobic and anaerobic fluidized bed bioreactors

have been developed for use in waste treatment.

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FLUIDIZED BED BIOREACTOR
Advantages Disadvantages

  Uniform Temperature  Increased Reactor Vessel

Gradients Size
 Ability to Operate  Pumping Requirements
Reactor in Continuous and Pressure Drop
State   Particle Entrainment

 Lack of Current
Understanding

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PACKED BED BIOREACTOR
 A bed of solid particles,
with biocatalysts on or
within the matrix of
solids, packed in a
column constitutes a
packed bed bioreactor.

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The solids used may be porous or non-porous gels, and they
may be compressible or rigid in nature.
 The concentration of the nutrients (and therefore the products
formed) can be increased by increasing the flow rate of the
nutrient broth.
 A nutrient broth flows continuously over the immobilized
biocatalyst.
 The products obtained in the packed bed bioreactor are
released into the fluid and removed.
 However, these bioreactors are preferred for bioprocessing
technology involving product-inhibited reactions. The packed
bed bioreactors do not allow accumulation of the products to 34
any significant extent.
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PACKED BED BIOREACTOR
Advantages Disadvantages

 Higher conversion per unit  Undesired heat gradients,

mass of catalyst than other poor temperature control
catalytic reactors  Poor mixing, difficult to
 Low operating cost control the pH
 Continuous operation  Difficult to clean

 No moving parts  Undesirable side

 Catalyst stays in the reactor reactions 

 Reaction mixture/catalyst
separation is easy • 35
Effective at high
temperatures and pressures Bhakti Bapat
TRICKLE FLOW BIOREACTOR
 Trickle bed reactors are a class
of packed bed reactors in which
the medium flows (or trickles)
over the solid particles. 
 In these reactors, the particles
are not immersed in the liquid
 They are used widely in aerobic
treatment of sewage.
 They are used to polish effluent
from the activated sludge or
anaerobic digestion process and
for the nitrification of
ammonia.
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TRICKLE FLOW BIOREACTOR
Advantages

 Simple treatment system

 Effective in dealing with high organic concentration
although varying as per the media type in use
 High performance reliability

 Low power consumption

 Durable processing parts

 Moderate skill set and technical expertise

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PHOTO BIOREACTOR

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 These are the bioreactors specialised for fermentation that can be carried
out either by exposing to sunlight or artificial illumination.
 Certain important compounds are produced by employing photo-
bioreactors e.g., β-carotene, asthaxanthin.
 They are made up of glass or more commonly transparent plastic.

 The array of tubes or flat panels constitute light receiving systems (solar
receivers).
 The culture can be circulated through the solar receivers by methods
such as using centrifugal pumps or airlift pumps.
 It is essential that the cells are in continuous circulation without forming
sediments.
 Further adequate penetration of sunlight should be maintained.

 The tubes should also be cooled to prevent rise in temperature.

 Photo-bioreactors are usually operated in a continuous mode at a

temperature in the range of 25-40°C.
 Microalgae and cyanobacteria are normally used.

 The organisms grow during day light while the products are produced 40

during night.
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PHOTO BIOREACTOR
Advantages Disadvantages

  Higher productivity  Capital cost is very high.

 Large surface-to-volume  The productivity and
ratio Better control of gas production cost in some
transfer. enclosed photo bioreactor
 Reduction in evaporation systems are not much better
of growth medium. than those achievable in
 More uniform open-pond cultures.
 The technical difficulty in
temperature. 
sterilizing
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MEMBRANE BIOREACTOR

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 The possibility of retaining all bacteria and viruses results in a
sterile effluent, eliminating extensive disinfection that would be
required otherwise and eliminate the corresponding hazards
related to disinfection by products.
 Compact then the traditional activated sludge system

 Reduces cost of sludge handling

 Traditional strategies for fouling mitigation such as air sparging,

physical cleaning techniques (i.e backflushing and relaxation) and
chemical maintenance cleaning have been incorporated in most
MBR designs as a standard operating strategy to limit fouling.
 Also, Membrane fouling problems can lead to frequent cleaning
of the membranes, which stop operation and require clean water 43
and chemicals.
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MEMBRANE BIOREACTOR
Advantages Disadvantages

  Higher effluent quality  Capital cost is very high.

the other treatment  Membrane fouling, caking,
systems. pore blocking
 Disinfection not required 

 Compact systems

 Easy maintenance

 Higher nitrogen removal

rate
 Low sludge rate 44

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ROTARY BIOREACTOR

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 Bed of bioreactor is mixed either continuously or
intermittently and air is circulated through head space of the
bed.
 Consist of a cylindrical drum lying horizontally

 Drum is partially filled with abed of substrate and air is blown

through headspace.
 The drum rotates around the central axis to mix the bed.

 Intermittent mixing bioreactor operates like a tray bioreactor

during static period and like a continuous rotating bioreactor
during period of rotation.
 It is necessary to limit the height of substrate bed in order to
achieve good O₂ and CO₂
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 Might include the use of baffles

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ROTARY BIOREACTOR
Advantages Disadvantages

   High oxygen transfer.  Difficult to scale up

 Good mixing facilitates
better growth and impart
less hydrodynamic stress.

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 MIST BIOREACTOR
 This is suitable for hairy root
cultivation of plant cells. 
 Static root mass is contained
in a chamber that is mostly
empty.
 Nutrients are supplied as mist
of fine droplets suspended in
circulating air currents that
penetrates the spaces between
the roots.

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MIST BIOREACTOR
Advantages Disadvantages

  High oxygen transfer.  Mesh trays and cylindrical

 Good mixing facilitates stainless steel meshes are
better growth and impart required
less hydrodynamic stress.

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