Different Types of Fermentors / Bioreactors

The heart of the fermentation or bioprocess technology is the Fermentor or Bioreactor. A

bioreactor is basically a device in which the organisms are cultivated to form the desired products.
it is a containment system designed to give right environment for optimal growth and metabolic
activity of the organism.
A fermentor usually refers to the containment system for the cultivation of prokaryotic cells, while
a bioreactor grows the eukaryotic cells (mammalian, insect cells, etc).

Types of Bioreactor

1. Continuous Stirred Tank Bioreactor

2. Airlift Bioreactor
3. Fluidized Bed Bioreactor
4. Packed Bed Bioreactor
5. Photobioreactor
6. Membrane Bioreactor
Continous Stirred Tank Bioreactor
In Continous Stirred Tank Bioreactor, the contents of the vessel no longer vary with time, this
applies to the hold up of micro-organisms and the concentration of the components of the medium
in the fermentor.Steady state conditions can be achieved by
either Chemostatic or Turbidostatic principles.The former involves the adjustment of the flow
rate of the fermentor to an appropriate and constant value and allowing the micro-organisms,
substrates and biochemical product concentration to attain their natural levels. The turbidostat
requires an experimental determination of the turbidity (ie, indirect measurement of microbial
concentration). This thus used to control the flow rate. Both these methods have been employed in
practice, though the former is obviously the simpler from every view point.
The most successful continuous systems to date have been those employing yeasts and bacteria, in
which the desired products are the cells or primary metabolites, compounds that form the chemical
'inventory' of a microbe, (e.g. enzymes and amino acids), or some product clearly associated with
growth or energy producing mechanisms (e.g. the production of alcohol).
The most widely used continous process based on CSTF (Continous Stirred Tank Fermentor) is
the activated sludge process used in waste water treatment industry.
In continuous processing the autocatalytic (a reaction in which one of the products of the reaction
increases the overall rate of a reaction) nature of microbiological reactions takes on a further
significance. This is because the presence of one of the products, additional micro-organisms,
enhances the overall rate of reaction. In the absence of micro-organisms no reaction can take
place. Therefore, it is essential to retain at least a portion within the fermentor. It follows that if the
flow rate is raised to a high value, then all the micro-organisms will be swept from the fermentor,
and the conversion will cease. This phenomenon is commonly known as 'Wash-out'. if micro-
organisms are fed to the fermentor simultaneously with the substrate feed, the problems
associated with wash-out are abated, and the reaction proceeds normally.

Advantages of Stirred Tank Bioreactor

1. Continuous operation
2. Good temperature control
3. Easily adapts to two phase runs
4. Good control
5. Simplicity of construction
6. Low operating (labor) cost
7. Easy to clean

Airlift Bioreactor
This kind of fermenter works on the principle of an air lift pump. It is of two kinds:
1. Internal loop type
2. External loop type.
The reactor's volume is determined by its capacity, kinetic data, and specific growth rate of the
organism used. The rate of airflow of the reactor depends on the volumetric mass transfer
coefficient in the reactor system. It is a uniform cylindrical cross type and has an internal loop or
external loop riser configuration, diverging converging. The external loop riser configuration is
adjustable and the change in the configuration improves the O2 transfer rate vis-a-vis mass
transfer coefficient for a particular rate of airflow. This helps provide required particular dissolved
O2 concentration for specific microbial system. This reactor reduces the operating cost
for pumping air through the bioreactor.
Advantages:
 Simple design with no moving parts or agitator for less maintenance, less risk of defects.
 Easier sterilization (no agitator shaft parts)
 Low Energy requirement vs stirred tank: Obviously doesn’t need the energy for the
moving parts (agitator shaft).
 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. It is difference to the Stirred tank
bioreactor that needs the heat coat or plate surrounding the tank to make warm bioreactor.
It is clear enough that the Airlift bioreactor has greater heat-removal compare to Stirred
tank.
 Very low cost
Fluidized Bed Bioreactor
This is a characteristic of beds of regular particles suspended in an up flowing liquid stream.
If an additional gas phase is involved, there is a tendency for the particles in the bed to become
less evenly distributed.
There are two important features of the beds of mixed particle sizes:
(i) The increase in porosity from the bottom to the top of the bed, and
(ii) The decreased particle movement when compared with beds containing particles of constant
size.
Since porosity or voidage is a measure of the local free space within a bed, it also represents a
measure of the microbial hold-up when expressed as wet volume per unit bed volume. Thus, a
variation in microbial hold-up is to be expected within a 'fluidised bed' fermentor on fluidisation,
the smaller particles rise relative to the larger particles, and produce a situation where the smaller
particles are at the top and the larger particles are at the bottom of the bed.
As the smaller particles have the lowest settling velocity, the bed arranges itself, so that the
smaller particles may be in the region of the highest porosity and the lowest linear velocity. The
tower fermentor (developed for the continuous production of beer) is based upon these general
principles (Ault et al, 1969). In this process yeast flocs are maintained in suspension by the
upward movement of the nutrient medium. Moreover, any entrained particles are returned by
means of a sedimentation device at the top of the tower.
Essentially, the fermentor consists of a vertical cylinder with an aspect ratio (length to diameter) of
approximately 10:1 At the top of the tower a separator is provided to induce the gas bubbles
produced by the reaction, to coalesce and escape from the liquid phase.
Within the separator there is a quiescent lone, free of the rising gas, so that the yeast may settle
and return to the main body of the tower, and clear beer can be removed. A flocculent yeast (i.e. a
yeast capable of achieving relatively large floc sizes) is essential for an alcoholic fermentation in a
PBP at acceptable flow rates, otherwise a large proportion of the yeast would be washed out. As a
result of this, an insufficient yeast concentration is maintained. A mean yeast concentration of 25
% by weight (expressed as centrifuged wet weight) is typical with values as high as 30-35%
by weight at the bottom of the tower, and as low as 5-10% by weight at the top.
A significant feature of the tower is the progressive and continuous fall in the specific gravity of
the nutrient medium between the bottom and the top of the tower. There is an initial rapid fall at
the bottom of the tower. It is followed by a slower fall over the middle and the top of the tower.
This gradual fall in the specific gravity is due to the fermentation of the sugars.
Advantages of Fluidized Bed Reactor:
 Uniform Particle Mixing: Due to the intrinsic fluid-like behavior of the solid material,
fluidized beds do not 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 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.
 Ability to Operate Reactor in Continuous State: The fluidized bed nature of these reactors
allows for the ability to continuously withdraw product and introduce new reactants into
the reaction vessel. Operating at a continuos process state allows manufacturers to
produce their various products more efficiently due to the removal of startup conditions in
batch process.

Photobioreactor
Advantages of Photobioreactor
 Cultivation of algae is in controlled circumstances, hence potential for much higher
productivity
 Large surface-to-volume ratio. PBRs offer maximum efficiency in using light and
therefore greatly improve productivity. Typically the culture density of algae produced is
10 to 20 times greater than bag culture in which algaeculture is done in bags – and can be
even greater.
 Better control of gas transfer.
 Reduction in evaporation of growth medium.
 More uniform temperature.
 Better protection from outside contamination.
 Space saving – Can be mounted vertically, horizontally or at an angle, indoors or
outdoors.
 Reduced Fouling – Recently available tube self cleaning mechanisms can dramatically
reduce fouling.
Membrane Bioreactor
Membrane bioreactors successfully applied to various microbial bioconversions such as alcoholic
fermentation, solvents, organic acid production, waste water treatment, etc.
In membrane bioreactor the soluble enzyme and substrate are introduced on one side of ultrafilter
membrane by means of a pump. product is forced out through the membrane. membrane holds
back the enzyme. good mixing in the reactor can be achieved by using a stirrer.
The most widely used membrane materials includes polysulfonte, polyamide and cellulose acetate.
Advantages of Membrane Bioreactor
1. The loss of enzyme is reduced.
2. Enzyme lost by denaturation can be make up by periodic addition of enzyme.
3. Substrate and enzyme can be easily replaced.
Additional notes
1. Continuous Stirred Tank Bioreactors:
A continuous stirred tank bioreactor consists of a cylindrical vessel with motor driven central shaft
that supports one or more agitators (impellers). The shaft is fitted at the bottom of the bioreactor.
The number of impellers is variable and depends on the size of the bioreactor i.e., height to
diameter ratio, referred to as aspect ratio.
The aspect ratio of a stirred tank bioreactor is usually between 3-5. However, for animal cell
culture applications, the aspect ratio is less than 2. The diameter of the impeller is usually 1/3 rd of
the vessel diameter. The distance between two impellers is approximately 1.2 impeller diameter.
Different types of impellers (Rustom disc, concave bladed, marine propeller etc.) are in use.
In stirred tank bioreactors or in short stirred tank reactors (STRs), the air is added to the culture
medium under pressure through a device called sparger. The sparger may be a ring with many
holes or a tube with a single orifice. The sparger along with impellers (agitators) enables better gas
distribution system throughout the vessel.
The bubbles generated by sparger are broken down to smaller ones by impellers and dispersed
throughout the medium. This enables the creation of a uniform and homogeneous environment
throughout the bioreactor.
Advantages of STRs:
There are many advantages of STRs over other types. These include the efficient gas transfer to
growing cells, good mixing of the contents and flexible operating conditions, besides the
commercial availability of the bioreactors.
2. Bubble Column Bioreactors:
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 —O2 transfer, mixing. The bubble column bioreactors may be fitted with
perforated plates to improve performance. The vessel used for bubble column bioreactors is
usually cylindrical with an aspect ratio of 4-6 (i.e., height to diameter ratio).
3. 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. 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.
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 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 circulation.
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.

Tower bioreactors:
A pressure-cycle fermenter with large dimensions constitutes a tower bioreactor. A high
hydrostatic pressure generated at the bottom of the reactor increases the solubility of O 2 in the
medium. At the top of the riser, (with expanded top) reduces pressure and facilitates expulsion of
CO2. The medium flows back in the down comer and completes the cycle. The advantage with
tower bioreactor is that it has high aeration capacities without having moving parts.
4. Fluidized Bed Bioreactors:
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. These bioreactors are suitable for use to carry out reactions involving fluid suspended
biocatalysts such as immobilized enzymes, immobilized cells, and microbial flocs.

For an efficient operation of fluidized beds, gas is spared to create a suitable gas-liquid-solid fluid
bed. 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.
5. Packed Bed Bioreactors:
A bed of solid particles, with biocatalysts on or within the matrix of solids, packed in a column
constitutes a packed bed bioreactor. The solids used may be porous or non-porous gels, and they
may be compressible or rigid in nature. A nutrient broth flows continuously over the immobilised
biocatalyst. The products obtained in the packed bed bioreactor are released into the fluid and
removed. While the flow of the fluid can be upward or downward, down flow under gravity is
preferred.
The concentration of the nutrients (and therefore the products formed) can be increased by
increasing the flow rate of the nutrient broth. Because of poor mixing, it is rather difficult to
control the pH of packed bed bioreactors by the addition of acid or alkali. However, these
bioreactors are preferred for bioprocessing technology involving product-inhibited reactions. The
packed bed bioreactors do not allow accumulation of the products to any significant extent.
6. Photo-Bioreactors:
These are the bioreactors specialised for fermentation that can be carried out either by exposing to
sunlight or artificial illumination. Since artificial illumination is expensive, only the outdoor
photo-bioreactors are preferred. Certain important compounds are produced by employing photo-
bioreactors e.g., p-carotene, asthaxanthin.
The different types of photo-bioreactors are depicted in Fig. 19.4. 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 during night.
Sterilization of media and air for bioreactors
Sterilization of Culture Media and Gases:
For successful fermentation, it is absolutely essential to ensure:
a. Sterility of the media containing the nutrients.
b. Sterility of incoming and outgoing air.
c. Sterility of the bioreactor.
d. Prevention of contamination during fermentation.
A bioreactor can be sterilized by destroying the organisms by heat/chemicals/radiation or
sometimes by physical procedures such as filtration.
Methods for Sterilization of Media and Air (With Diagram)

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Read this article to learn about the various methods for sterilization of media and air.
Sterilization of Culture Media and Gases:
For successful fermentation, it is absolutely essential to ensure:
a. Sterility of the media containing the nutrients.
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b. Sterility of incoming and outgoing air.
c. Sterility of the bioreactor.
d. Prevention of contamination during fermentation.
A bioreactor can be sterilized by destroying the organisms by heat/chemicals/radiation or
sometimes by physical procedures such as filtration.
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Sterilization of media and air are discussed below:
1. Sterilization of Culture Media:
The constituents of culture media, water and containers contribute to the contamination by
vegetative cells and spores. The media must be free from contamination before use in
fermentation. Sterilization of the media is most commonly achieved by applying heat and to a
lesser extent by other means (physical methods, chemical treatment, and radiation).
Heat sterilization:
Heat is the most widely used sterilization technique. The quality and quantity of contamination
(i.e., the type and load of microorganisms), composition of the media and its pH and size of the
suspended particles are the important factors that influence the success of heat sterilization.
In general, vegetative cells are destroyed at lower temperature in a short time (around 60°C in 5-
10 minutes). However, destruction of spores requires higher temperature and relatively longer
time (around 80°C for 15-20 minutes). Spores of Bacillus stearothermophilus are the most heat
resistant. In fact, this organism is exploited for testing the sterility of fermentation equipment.
Physical methods:
The physical methods such as filtration, centrifugation, and adsorption (to ion-exchangers or
activated carbon) are in use. Among these, filtration is most widely used. Certain constituents
(vitamins, blood components, antibiotics) of culture media are heat labile and therefore, are
destroyed by heat sterilization. Such components of the medium are completely dissolved
(absolutely essential or else they will be removed along with microorganisms) and then subjected
to filter sterilization.
There are a couple of limitations of filtration technique:
1. Application of high pressure in filtration is unsuitable for industries.
2. Some of the media components may be lost form the media during filtration.
Sometimes, a combination of filtration and heat sterilization are applied. For instance, the water
used for media preparation is filtered while concentrated nutrient solution is subjected to heat
sterilization. The filtered water is now added for appropriate dilution of the media. The chemical
methods (by using disinfectants) and radiation procedures (by using UV rays, y rays, X-rays) are
not commonly used for media sterilization.
Batch sterilization:
The culture media are subjected to sterilization at 121°C in batch volumes, in the bioreactor. Batch
sterilization can be done by injecting the steam into the medium (direct method) or injecting the
steam into interior coils (indirect method). For the direct batch sterilization, the steam should be
pure, and free from all chemical additives (that usually come from steam manufacturing process).
There are two disadvantages of batch sterilization:
1. Damage to culture media:
Alteration in nutrients, change in pH and discolouration of the culture media are common.
2. High energy consumption:
It takes a few hours (2-4 hrs.) for the entire contents of the bioreactor to attain the requisite
temperature (i.e. 120°C). Another 20-60 minutes for the actual process of sterilization, followed
by cooling for 1-2 hours. All this process involves wastage of energy, and therefore batch
sterilization is quite costly.
Continuous sterilization:
Continuous sterilization is carried out at 140°C for a very short period of time ranging from 30 to
120 seconds. (This is in contrast to the batch fermentation done at 121°C for 20-60 minutes). This
is based on the principle that the time required for killing microorganisms is much shorter at
higher temperature. Continuous sterilization is carried out by directly injecting the steam or by
means of heat exchangers.
In either case, the temperature is very quickly raised to 140°C and maintained for 30- 120 seconds.
The stages of continuous sterilization process and the corresponding temperatures are depicted in
figure below. The different stages are— exchanger, heater, heat maintenance unit, recovery of
residual heat, cooling and fermenter.

In the continuous sterilization process, 3 types of heat exchangers are used. The first heat
exchanger raises temperature to 90-1 20°C within 20-30 seconds. The second exchanger further
raises temperature to 140°C and maintains for 30-120 seconds. The third heat exchanger brings
down the temperature by cooling in the next 20-30 seconds. The actual time required for
sterilization depends on the size of the suspended particles. The bigger is the size, the more is the
time required.
The main advantage with continuous sterilization is that about 80-90% of the energy is conserved.
The limitation however, is that certain compounds in the medium precipitate (e.g., calcium
phosphate, calcium oxalate) due to very high temperature differences that occur in a very short
time between sterilization and cooling. The starch-containing culture media becomes viscous in
continuous sterilization and therefore is not used.
2. Sterilization of Air:
In general, the industrial fermentations are carried out under vigorous and continuous aeration. For
an effective fermentation, the air should be completely sterile, and free from all microorganisms
and suspended particles. There is a wide variation in the quantity of suspended particles and
microbes in the atmospheric outdoor air.
The microorganisms may range from 10-2,000/m 3 while the suspended particles may be 20-
100,00/ m3. Among the microorganisms present in the air, the fungal spores (50%) and Gram-
negative bacteria (40%) dominate. Air or other gases can be sterilized by filtration, heat, UV
radiation and gas scrubbing. Among these, heat and filtration are most commonly used.
(a) Air sterilization by heat:
In the early years, air was passed over electrically heated elements and sterilized. But this is quite
expense, hence not in use these days.
(b) Air sterilization by filtration:
Filtration of air is the most commonly used sterilization in fermentation industries.
Depth filters:
When the air is passed through a glass wool containing depth filters the particles are trapped and
removed. This filtration technique primarily involves physical effects such as inertia, blocking,
gravity, electrostatic attraction and diffusion. Glass wool filters can be subjected to steam
sterilization and reused. But there is a limitation in their reuse since glass wool shrinks and
solidifies on steam sterilization. In recent years, glass fiber filter cartridges (that do not have the
limitations of glass wool filter) are being used.

Membrane cartridge filters:

These are removable pleated membrane filters made up of cellulose ester, nylon or polysulfone.
Membrane cartridge filters are smaller in size, simpler for operation and replacement. The most
important limitation of air sterilization is that there is no filter that can remove bacteriophages.
Bacteriophages are capable of crippling the industrial fermentation. e.g., bacteriophages interfere
in the production of glutamic acid by Corynebacterium glutamicum.