BIO

REACTOR

Dr. Suchitra Patil

 INTRODUCTION
• Bioreactor – can be described as a vessel which has provision
of cell cultivation under sterile condition & control of
environmental conditions e.g., pH, Temperature, Dissolved
oxygen etc.

• It can be used for the cultivation of microbial plant or animal

cells.

• This process can either be aerobic or anaerobic.

• The bioreactors are commonly cylindrical, ranging in size

from litres to cubic metres and are made of stainless steel.
 BIOREACTOR DESIGN

• The design and mode of operation of a bioreactor are

based on the production of an organism, optimum
conditions required for desired product formation,
product value, and its scale of production.

• A good bioreactor design will help to improve

productivity and provide higher quality products at
lower prices.
A bioreactor is a device that consists of various features such as -

• agitator system,
• oxygen delivery system,
• foam control system,
• temperature control system
• pH control system,
• sampling ports,
• cleaning system
• sterilization system,
• system for charging & emptying the reactor.
The material used for the construction of a
bioreactor must have the following important
properties:

• It should not be corrosive.

• It should not add any toxic substances to the
fermentation media.
• It should tolerate the steam sterilization process.
• It should be able to tolerate high pressure and
resist pH changes.
The sizes of the bioreactor vary widely depending on
the application -

• Some bioreactors are designed for small scale

fermenters and some for large scale industrial
applications.

• shake flask (100-1000 ml)

• laboratory-scale fermenter (1 – 50 L)
• pilot level (0.3 – 10 m3)
• plant scale (2 – 500 m3)
 BIOREACTOR PRINCIPLE

• The bioreactor is the heart of any biochemical process as it

provides an environment for microorganisms to obtain optimal
growth and produce metabolites for the biotransformation
and bioconversion of substrates into desirable products.

• The reactors can be engineered or manufactured based on the

growth requirements of the organisms used.

• They can be used for the production of various enzymes and

other bio-catalysis processes.
PARTS OF THE BIOREACTOR AND THEIR FUNCTION

• These reactors have been designed to maintain certain

parameters like flow rates, aeration, temperature, pH, foam
control, and agitation rate.

• The number of parameters that can be monitored and

controlled is limited by the number of sensors and control
elements incorporated into a given bioreactor.

• Other factors should be kept in mind before designing a

fermenter as described below and demonstrated in the
figure below.
 DIAGRAM
OF
BIOREACTOR
DIAGRAM OF A TYPICAL BIOREACTOR
 SPECIFICATIONS OF A BIOREACTOR
• A typical bioreactor consists of following parts:

1. Fermenter Vessel
• A fermenter is a large cylinder closed at the top and bottom
connected with various pipes and valves.
• The vessel is designed in such a way that it allows to work under
controlled conditions.
• Glass and stainless steels are two types of fermenter vessels used.
• The glass vessel is usually used in small-scale industries. It is non-
toxic and corrosion-proof.
• Stainless steel vessel is used in large scale industries. It can resist
pressure and corrosion.
Images of Fermenter Vessel
2. Heating and Cooling Apparatus

• The fermentor vessel’s exterior is fitted with a cooling jacket

that seals the vessel and provides cooling water.

• Thermostatically controlled baths or internal coils are

generally used to provide heat while silicone jackets are used
to remove excess heat.

• A cooling jacket is necessary for sterilization of the nutrient

medium and removal of the heat generated during
fermentation in the fermentor.
Image of cooling jacket
3. Aeration System

• An aeration system is one of the very important parts of a

fermentor.

• It is important to choose a good aeration system to ensure

proper aeration and oxygen availability throughout the culture.

• It contains two separate aeration devices (sparger and impeller)

to ensure proper aeration in a fermentor.
The stirring accomplishes two things:

• It helps to mix the gas bubbles through the liquid culture

medium.

• It helps to mix the microbial cells through the liquid culture

medium which ensures the uniform access of microbial cells
to the nutrients.
4. Sealing Assembly

• The sealing assembly is used for the sealing of the stirrer

shaft to offer proper agitation.

There are three types of sealing assembly in the fermenter:

• Packed gland seal

• Mechanical seal
• Magnetic drives
Images of mechanical seals of fermenter
5. Baffles

• The baffles are incorporated into fermenters to prevent a

vortex and improve aeration in the fermenters.

• It consists of metal strips attached radially to the wall.

Images of baffles
6. Impeller

• Impellers are used to provide uniform suspension of microbial

cells in different nutrient mediums.
• They are made up of impeller blades attached to a motor on
the lid.
• Impeller blades play an important role in reducing the size of
air bubbles and distribute them uniformly into the
fermentation media.
Variable impellers are used in the fermenters and are classified
as follows -
• Disc turbines
• Variable pitch open turbine
 TYPES OF IMPELLER

1. Open type

2. Partially open/closed

3. Closed type
 IMPELLERS
 Impeller agitators divided into two classes.
 AXIAL-FLOW IMPELLERS
 Those impellers that generate currents parallel with the
axis of the impeller shaft are called Axial-Flow
Impellers.

Axial-flow impeller
 RADIAL-FLOW
IMPELLERS
 Those impellers that generate currents an a radial or
tangential direction are called radial flow
impellers.

Radial-flow impellers
 TYPES OF
IMPELLERS
Three main types of impellers for low-to moderate
viscosity
 liquids are.
 PROPELLERS
 A propeller is an axial-flow, high speed impeller
for liquids of low viscosity.

 Small propellers turn at full speed either 1,150 or

1,750 rpm.

 Larger propellers turn at 400 to 800 rpm.

 TURBINES
There are 4 types of turbine impeller

 The straight-blade turbine

 Disc turbine

 Concave blade CD-6 disc turbine

 Pitched-blade turbine
 The straight-blade turbine
 The simple pushes the liquid
radially and tangentially with
almost no vertical motion at the
impeller.

 The current it generate travel

outward to the vessel wall and
then flow either upward or
downward.

 Such impellers sometimes called

straight-blade turbine
p a dd le s .
 Disc turbine
 The disc turbine with multiple straight blades mounded
on a horizontal disc.

 Useful for dispersing a gas in a liquid.

 Concave blade CD-6 disc
turbine

 It is also used for gas

dispersion.
 Consists of six concave
blades.
 Pitched-blade turbine

 Used when good overall

circulation important.

 Because it provides some axial

flow in addition to the radial
flow.
 HIGH EFFICENCY IMPELLERS
• The high efficiency impeller E-3 has
three slanted blades that are crimped to
decrease the blade angle near the tip.

• The A3 1 0 fluid fo il im p e lle r uses air

foil shaped blades with taper so that they
are narrower at the tip than their edges.

• Used to mix low or moderate viscosity

liquids.
7. Sparger

• A sparger is a system used for introducing sterile air to a

fermentation vessel.
• It helps in providing proper aeration to the vessel.
• The sparger pipes contain small holes of about 5-10 mm,
through which pressurized air is released.

Three types of sparger are used

• Porous sparger
• Nozzle sparger
• Combined sparger–agitator
Images of sparger
Stainless steel Sparger Porous Sparger
8. Feed Ports

• They are used to add nutrients and acid/alkali to the

fermentor.

• Feed ports are tubes made up of silicone.

• In-situ sterilization is performed before the removal or

addition of the products.
Images of feed port
9. Foam-Control

• The level of foam in the vessel must be minimized to avoid

contamination.

• Foam is controlled by two units foam sensing and a control

unit.

• A foam-controlling device is mounted on top of the

fermentor, with an inlet into the fermentor.
Images of foam control
10. Valves

• Valves are used in the fermentor to control the movement of

liquid in the vessel.

There are around five types of valves are used -

• globe valve
• butterfly valve
• ball valve
• diaphragm valve
• safety valve is built-in in the air and pipe layout to operate
under pressure
Images of valve
11. Controlling Devices for Environmental Factors

• A variety of devices are utilized to control environmental

elements like temperature, oxygen concentration, pH,
cell mass, essential nutrient levels, and product
concentration.
12. Use of Computer in Fermenter

• For an efficient process, monitoring, and data collecting,

fermentors are generally coupled with modern
automated and semi-automated computers and
databases.
Desktop fermenter and computer
 CONCLUSION

A variety of bioreactor types providing growth and

expression of bioactive substances are available today.

An improved understanding of the interactions between

cultivated cells, product formation and the specific designs
for different bioreactor types will enhance and sustain high
productivity and also reduce the process costs.
Types of Bioreactors
 BIOREACTORS TYPES
1. Continuous Stirred Tank Bioreactors
2. Bubble Column Bioreactors
3. Airlift Bioreactors
4. Fluidized Bed Bioreactors
5. Packed Bed Bioreactors
6. Photo-Bioreactors
 IMMOBILIZED
CELL
BIOREACTORS
OR
FLUIDIZED BED
BIOREACTORS
 FLUIDIZED BED BIOREACTORS /
IMMOBILIZED CELL BIOREACTORS
• 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.

• These bioreactors are suitable for use to carry out reactions

involving fluid suspended biocatalysts such as immobilized enzymes,
immobilized cells, and microbial flocs.

• An upflowing stream of liquid is used to suspend or “fluidize” the

relatively dense solids.
Basic diagram of a fluidized bed reactor
 The reactor is expanded in the top to reduce the
superficial velocity of the liquid to a value below that
needed to keep the solids in suspension.

 Consequently solids sediment in the expanded zone and

drop back into the narrower reactor column.

 Hence the solids are retained in the column and liquid

flows out.
 A liquid fluidized bed may be sparged with air or any
gas to produce a gas-liquid-solid fluidised bed.

 If the solid particles are too light they may have to be

artificially weighted, for example by embedding
stainless steel balls.

 A high density of solids improve solid-liquid mass

transfer and denser solids are easier to sediment.
 But the density should not be too high, otherwise
fluidization itself may be difficult.

 Even with lighter solid particles, superficial velocity

needed to suspend the solids may cause the liquid to
leave from the reactor too quickly.

 This leads to insufficient solid liquid contact time for the

reaction and liquid may have to be recycled to obtain
sufficiently long cumulative time with the biocatalyst.
 The minimum fluidization velocity –the superficial liquid
velocity needed to just suspend the solids from a settled
state depends on

 Density difference between two phases

 Shape and diameter of the particles

 Viscosity of the liquid.

 Advantages

Uniform Particle Mixing:

Due to the intrinsic fluid-like behavior of the solid material, fluidized beds do not
1 experience poor mixing as in packed beds. This complete mixing allows for a
uniform product that can often be hard to achieve in other reactor designs. The
elimination of radial and axial concentration gradients also allows for better fluid-
solid contact, which is essential for reaction efficiency and quality.

Uniform Temperature Gradients:

Many chemical reactions require the addition or removal of heat. Local hot or
2 cold spots within the reaction bed, often a problem in packed beds, are avoided in
a fluidized situation such as an FBR. In other reactor types, these local
temperature differences, especially hotspots, can result in product degradation.
Thus FBRs are well suited to exothermic reactions. Researchers have also learned
that the bed-to-surface heat transfer coefficients for FBRs are high.
CONTINUED…

Ability to Operate Reactor in Continuous State:

The fluidized bed nature of these reactors allows for the ability
3 to continuously withdraw product and introduce new reactants
into the reaction vessel. Operating at a continuous process state
allows manufacturers to produce their various products more
efficiently due to the removal of startup conditions in batch
processes.

USEFUL IF THE REACTION INVOLVES THE

4 UTILISATION OR RELEASE OF GASEOUS MATERIAL.
 DISADVANTA
GES

Increased Reactor Vessel Size:

1 Because of the expansion of the bed materials in the reactor, a larger vessel is
often required than that for a packed bed reactor. This larger vessel means that
more must be spent on initial capital costs.

Pumping Requirements and Pressure Drop:

The requirement for the fluid to suspend the solid material necessitates
2 that a higher fluid velocity is attained in the reactor. In order to achieve
this, more pumping power and thus higher energy costs are needed. In
addition, the pressure drop associated with deep beds also requires
additional pumping power.
 CONTINUED…

Particle Entrainment:
The high gas velocities present in this style of reactor often result in fine
particles becoming entrained in the fluid.
3 These captured particles are then carried out of the reactor with the fluid,
where they must be separated. This can be a very difficult and expensive
problem to address depending on the design and function of the reactor. This
may often continue to be a problem even with other entrainment reducing
technologies.

4 DIFFICULTY IN SCALLING-UP THESE REACTIONS.

APPLICATIONS OF FLUIDIZED BED
BIOREACTOR
IN WASTE WATER TREATMENT:

• The use of fluidized bed reactor in wastewater treatment,

represents a unique opportunity for cost-effective treatment of
wastewater containing recalcitrant pollutants (compounds that are
slowly biodegradable or non- biodegradable are known as
recalcitrant compounds, and range from simple halogenated
hydrocarbons to complex polymers)

• Although the application of fluidized bed reactor in biological

wastewater treatment is well established with many large-scale
plants in existence, its application in advanced oxidation processes
is mostly at laboratory-scale.
A major application of FBBs is fluidized catalytic
cracking which was developed in 1940s.

Anaerobic fluidized-bed platform has been

tested as an anaerobic platform for the high
strength and high solids waste streams such as
the municipal sludge's and corn ethanol thin
stillage.
A photograph of a
fluidized bed bioreactor
(inside volume 450 cm3)
used to produce butanol
from whey permeate
using C. acetobutylicum
P262.
Images of a fluidized bed reactor
 AIRLIFT
BIOREACTO
RS
 AIRLIFT BIOREACTORS
• In the airlift bioreactors, the medium of the vessel is divided
into two interconnected zones by means of a baffle or draft
tube.

• In one of the two zones referred to a riser, the air/gas is

pumped. The other zone that receives no gas is the down
comer.

• The dispersion flows up the riser zone while the down flow
occurs in the down comer.
BASIC IMAGE OF AIRLIFT BIOREACTOR
 TYPES OF AIRLIFT BIOREACTORS
There are two types of 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 contents is better compared to bubble columns.
Schematic of airlift bioreactor with (a) external recirculation
and (b) internal recirculation
IMAGES OF AIRLIFT BIOREACTORS
 TWO-STAGE AIRLIFT BIOREACTORS
• 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
proper takes place in the second reactor.
IMAGE OF TWO-STAGE AIRLIFT
BIOREACTORS
1. Highly energy efficient and productivities are comparable
to those of stirred tank bioreactors.
2. Simple design with no moving parts or agitator for less
maintenance, less risk of defects.
3. Easier sterilization (no agitator shaft parts)
4. Low Energy requirement vs. stirred tank Obviously doesn’t need
the energy for the moving parts (agitator shaft).
5. Greater heat-removal vs. stirred tank
At the Airlift bioreactor it doesn’t need the heat plate to control
the temperature, because the Draught-Tube which is inside the
bioreactor can be designed to serve as internal heat exchanger.
1. The agitation on the Airlift bioreactor is controlled by the
supply air to adjust the supply air then the higher pressure
needed.

2. The higher pressure of air needed then more energy

consumption needed and more cost must pay.

3. Inefficient break the foam when foaming occurs.

4. No bubbles breaker, There are no blades that used as a breaker

the bubbles which produced from the air supply (sparger).
1. The reactor is commonly used in the culture of shear
sensitive organisms.

2. Airlift bioreactors are commonly employed for aerobic

bioprocessing technology. They ensure a controlled
liquid flow in a recycle system by pumping.

3. Due to high efficiency, airlift bioreactors are sometimes

preferred e.g., methanol production, waste water treatment,
single-cell protein production.
Which type of reactor is the world's
largest industrial fermenter?
 A STR can never be very large because of the heat-
exchanger, mixing and aeration problems.

 The world's largest fermenter is Air-lift Bioreactor!

The fermenter is 200' high and 25 ft diam. The first
photo shows the fermenter being transported on
vehicles with tank treads.
 The next photo shows the fermenter being raised into
position at the ICI, Ltd. factory, Billingham, UK.
 MODES
OF
FERMENTATION
 Modes of
fermentation:

• Batch fermentation
• Continuous fermentation
• Fed batch fermentation
1. Batch culture technique
• It is also called as closed system of cultivation.

•In this technique at first nutrient solution is prepared and

it is inoculated with inoculum and then nothing is added in
the fermentation tank except aeration.

•In batch culture, neither fresh medium is added nor

used up media is removed from the cultivation vessel.
Therefore volume of culture remains same.
•Since fresh media is not added during the course of
incubation, concentration of nutrition decreases
continuously.

•Furthermore various toxic metabolites also accumulates in

the culture vessel.

•Therefore batch culture technique gives characteristics

growth curve with lag phase, log phase, stationary phase and
decline phase.
Lag phase: microbial population remains constant as
there is no growth .However it is the period of intense
metabolic activity.

Exponential phase: cells divide with increasing frequency

till it reaches the maximum growth rate. At this point
logarithmic growth begins and cell numbers and cell
biomass increase at constant rate.
Stationary Phase
The specific growth rate of the microorganism continues
decelerating until the substrate is completely depleted.
Overall growth rate has declined to zero and there is no net
change in cell numbers/ biomass
i.e. rate of cell division equals rate of cell death.
Microorganisms are still metabolically active, metabolizing
intracellular storage compounds, utilizing
nutrients released from lysed cells and in certain cases
produce secondary metabolites.

Death rate: cells die at constant rate and undergone lysis.

2. Continuous culture technique
• It is also called as open system of cultivation.

• In this technique fresh sterile medium is added

continuously in the vessel and used up media with bacterial
culture is removed continuously at the same rate.

• So the volume and bacterial density remain same in the

cultivation vessel.
•In this technique, bacteria grow continuously in their log
phase. This type of growth is known as steady state growth.

•The cell density in continuous culture remains constant

and it is achieved by maintaining constant dilution and
flow rate.
Continuous culture apparatus
Microbial growth in a chemostat
3. Fed-batch culture
• It is also called as semi-closed system of cultivation.

•In this technique, at first nutrient media is prepared and

it is inoculated with culture organism and then incubated
for particulate time.

•During the course of incubation a particular nutrient is

added at intervals without removing the used up media
so the volume of culture increases continuously.
•Fed batch culture technique is applied in many types of
fermentation process.

•In fermentation some nutrient is very essential for the

process but when these nutrients are provided in higher
concentration in the culture they inhibit the growth of
bacteria ultimately ceasing the fermentation.

•Therefore such nutrients are kept in lower

concentration initially and it is added slowly and
continuously during the course of fermentation.
Difference b/w batch, continuous and fed batch
Characteristics Batch culture Fed-batch culture Continuous culture

Cultivation system Closed type Semi-closed type Open type

Addition of fresh nutrition No Yes Yes

Volume of culture Constant Increases Constant

Removal of wastes No No Yes

Chance of contamination minimum Intermediate Maximum

Lag, log, stationary and Lag, log , stationary and

Growth phase decline phase decline phase Lag and log phase

Log phase Shorter longer Longest and Continuous

Density of bacteria Change with time Change with time Remain same

Product yield Low Medium High