ASSIGNMENT ON MCB 432 (INDUSTRIAL MICROBIOLOGY)

FERMENTORS AND FERMENTOR OPERATION

Definition of a fermentor
The aerated stirred tank batch fermentor
Construction materials for fermentors
Aeration and agitation in a fermentor
Temperature control in a fermentor
Foam production and control
Foaming patterns and foam control
What are Fermentors?
Fermentors, also known as bioreactors, are sterilised and enclosed vessels that are used for the
growth of microorganisms under optimal conditions. The microorganisms can be grown in large
quantities to produce metabolites for commercial uses. Fermentors are equipped with special
components for heating, mixing, and aeration. Its volume can be as big as 500,000 litres for an
industrial scale, or as small as 1 litre for laboratory uses. The fermentor is a vessel or an
apparatus which is used for the fermentation activity of the microorganisms for the commercial
production of certain substances. There are different types of fermentors depending on the type
of product and the volume of the product to be produced. The common type of fermentors are
tower fermenter, bubble column, air-lift, fluid bed fermenter, etc.
A Fermentor is a device used to accomplish the fermentation process by using
microorganisms, and for this reason, it is also called “Biofermentor or Bioreactor“. It is
equipped with all the elements that are necessary to carry out the commercial production of
substances like antibiotics, enzymes, beverages etc. in many industries.
Fermentor is designed in such a way that a higher value of product could form by the
microorganisms by utilizing a small amount of media. It shows broad applicability in the food
industry, wastewater treatment etc. Fermentation is the bioprocessing of the raw substrates
through microbial biomass (mostly yeasts and bacteria).

Properties of a Fermentor
 It should be reliable for long-term operation.
 A fermentor should be capable of being operated aseptically or should provide sterile
conditions.
 The bioreactor provides adequate aeration and agitation for uniform mixing of the
contents in the vessel.
 It should consume less power.
 A fermentor must be equipped with controlling probes that can maintain the temperature,
pH, oxygen level etc.
 It facilitates the passage of inoculum and media into the vessel.
 A bioreactor does not allow excessive evaporation loss.
 It minimizes the labour input for the operation, harvesting, cleaning and maintenance.
Basic components are necessary for the construction of fermentor, which involves:
Top-plate: It is the cover that is generally made of stainless steel.
Inoculation pipe: It helps to port the inoculum inside the fermentor.
Drive motor: It drives the impeller shaft.
Impeller shaft: Holds the agitator centrally.
Impeller: Acts as an agitating device for mixing up the nutrients and microorganisms uniformly.
Stirrer: Mixes the gas bubbles throughout the liquid culture medium.
Baffle: Prevents the counterflow or vortex formation by breaking down the gas bubbles to
improve aeration efficiency.
Sparger: It supplies oxygen into the culture medium through the perforated tubes.
Drain point: Withdraws cells or medium for the continuous fermentation.
Cooling jacket: It is fitted externally to the fermentation vessel which allows the passage of
steam or cold water to balance the heat generated during the process.
THE AERATED STIRRED TANK BATCH FERMENTOR
The stirred tank bioreactor is a simple and widely used fermenter design that consists of
a cylindrical vessel with a stirrer. The design has been used extensively in all microbial
fermentation and has been the main system used in yeast fermentation in the brewing
industry for centuries.
 A continuous stirred tank bioreactor is made up of a cylindrical vessel with a central shaft
controlled by a motor that supports one or more agitators (impellers).
 The sparger, in combination with impellers (agitators), allows for improved gas
distribution throughout the vessel.
 A stirred tank bioreactor can be operated continuously in the fermentor, temperature
control is effortless, construction is cheap, easy to operate, resulting in low labor cost,
and it is easy to clean.
 It is the most common type of bioreactor used in industry.
It has various applications in Antibiotics, citric acid, Exopolysaccharides, cellulose, Chitinolytic
enzymes, Laccase, Xylanase, Pectic, and pectate lyase, Tissue mass culture, Lipase,
Polygalacturonases, Succinic acid,etc.
•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.
Its basic functions include:
 Homogenization
 Suspension of solid material
 Aeration to the medium
 Heat exchange
In stirred tank fermentor, rotating stirrer and baffle are found either at the top or the bottom. It
mainly uses the batch process of fermentation.
CONSTRUCTION MATERIALS FOR FEMENTORS
As fermentation required adequate aseptic conditions, for better yield of biomass or product, it is
important to select a material for the body of the fermenter, which restricts the chances of
contamination. Moreover, it needs to be non-toxic and corrosion free. Glass is a material that
provides a smooth surface inside the vessel and also non-toxic in nature. Apart from that, it is
corrosion-proof and due to the transparency, it is easy to examine the inside of the vessel. There
are mainly two types of glass fermenters
1. A glass vessel with a flat bottom and a top plate with a diameter of 60 cm
(approximately). Sterilization of this type of vessel is performed by general autoclaving.
Borosilicate battery jars were used as large glass vessels.
2. The second glass vessel contains stainless steel plates at the top and bottom of the glass
vessel. In situ sterilization is possible for this type of glass vessel. However, it is more
expensive than the glass top vessels due to the use of stainless steel plates.
The main disadvantage of glass vessels is that it is difficult to top to design a pilot-scale
fermenter with glass. It is difficult to handle glass as a pilot-scale fermenter. Therefore, another
non-toxic, corrosion-proof material, stainless steel, was used for pilot scale fermenter. According
to Americal Iron and Steel Institute, steel contains more than 4% chromium is standardized as
stainless steel. However, the minimum amount of chromium required to protect the steel from
corrosion depends on the corroding agent present in a specific environment. In a pilot-scale
fermenter normally the steel contains around 10-13% of chromium. In many cases nickel is also
mixed in high concentration with the chromium to make the steel more corrosion resistant and it
also provides engineering advantages. In this modern-day, stainless steel fermenters are mostly
used for industrial production. However, small scale production requires glass vessels.
AERATION AND AGITATION IN A FERMENTOR
The primary use of Aeration is to provide microorganisms in submerged culture with sufficient
oxygen for metabolic requirements. Agitation ensures that a uniform suspension of microbial
cells is achieved in a homogenous nutrient medium.
What is Aeration?
Aeration is the process where the supply of adequate oxygen to the microorganisms takes place
in a submerged culture. This increases the oxygen concentration in the culture. The device that
introduces air to the fermenter is called a sparger. During aeration, the air supply to the fermenter
should be as fine bubbles with a high surface area. This allows efficient oxygen transfer to the
microorganisms to promote fermentation. In some fermenters, aeration also provides agitation.
Therefore, a separate agitator will not be required for the fermenter with medium-level or low
viscous fluids.
The three types of aeration spargers are porous sparger, orifice sparger, and nozzle sparger. All
spargers should consist of air filters to prevent contamination. Porous spargers are used only in
lab-scale fermenters, while nozzle spargers are used in larger-scale fermenters. In medium-scale
fermenters, the orifice sparger is used for aeration.

What is Agitation?
Agitation is the process of uniform suspension of microbial cells to homogenize the nutrient
medium. The function of agitation is to facilitate bulk fluid and gas phase mixing, oxygen
transfer, air dispersion, heat transfer, and a uniform condition within the fermenter. The impeller
or agitator facilitates the agitation inside the fermenter.
There are four types of impellers: disc impeller, vanned disc impeller, open impeller, and marine
impeller. The disc impeller prevents the flooding caused by air bubbles due to the lack of
dispersion of air bubbles properly during aeration. Modern fermenters consist of a dual impeller
for agitation that provides dual agitation. One impeller is for gas dispersion, and the other
impeller is for circulation. Similar to aeration, agitation is a crucial step in fermentation to help
the fermenter to provide the required results.
What are the Similarities Between Aeration and Agitation?
 Aeration and agitation are two mechanical processes.
 They play a key role in microbial fermentation.
 Both provide optimum conditions for the growth of microorganisms.
 Moreover, both processes require machinery.
 Both aeration and agitation are controlled processes.
What is the Difference Between Aeration and Agitation?
Aeration uses a sparger apparatus, while agitation uses an impeller or an agitator blade during the
fermentation process. Thus, this is the key difference between aeration and agitation. Aeration is
the process that supplies adequate oxygen to microorganisms in a submerged culture, while
agitation is the uniform suspension of microbial cells to homogenize the nutrient medium.
Moreover, there are three key types of aeration systems, while there are four types of agitation
systems.
The key difference between aeration and agitation is that aeration is done using a sparger
apparatus in order to provide oxygen for growing cells, while agitation is done using an impeller
or an agitator blade in order to distribute oxygen and nutrients uniformly inside the bioreactor.
TEMPERATURE CONTROL IN A FERMENTOR
In case of fermentation, a temperature control helps to control the temperature at the optimum
level by removing or providing heat.
If the fermentation temperature is too cold then the yeast goes dormant and won’t complete the
fermentation. On the other hand, if the fermentation temperature is too hot the yeast will become
stressed, creating all sorts of byproducts that cause off flavours and fusel alcohols that taste like
solvents. Way too hot and the cell walls of the yeast become permeable and alcohol which is
toxic to yeast will kill the yeast.

The best way of controlling fermentation temperature will be dictated by ambient temperatures
and whether we need to heat or cool the fermenter in order to stay within the ideal temperature
range.
Cooling Fermentation
The most common need, particularly if fermenting an ale or wine in a centrally heated house is to
keep things cool.
Cool Area / Fermentation Chamber
The simplest way to do this is to keep the fermenter in a cool cupboard that isn’t directly heated.
What we do need to be aware of though is we maintain a consistent temperature throughout a 24-
hour period. Swings of temperature from warm to cool will stress the yeast.
Water Bath
Another way to control fermentation temperature is to immerse the fermenter in a water bath. A
builders trub is a cheap container for a water bath. The extra mass of the water bath takes longer
to heat and cool which prevents temperature swings throughout the day and night.
You can also add ice to the water bath which will keep the temperature of the fermenter lower
than ambient temperatures.
Temperature Controlled Refrigerator
If you have more funds and space available the most precise and efficient option is to convert an
unused fridge into a temperature-controlled fermentation chamber.
If you have a fridge large enough for your fermenter then a temperature controller can be used to
control the internal temperature of the refrigerator.
A temperature controller such as this one can be set to cycle the power on and off for the
refrigerator according to the temperature you set.
The added benefit of an automatic temperature controller attached to the fridge is you can also
add a heating device inside the refrigerator to have even more control and accuracy of the
fermentation temperature.

Heating Fermentation Temperatures

If you require warmer temperatures for your fermentation, such as fermenting in an unheated
garage then you will need a device to heat the fermenter.
Heating Belts
Heating belts, which are adjustable bands that plug into the mains to produce heat can be slipped
around the outside of the fermenter.
These heat belts produce a set amount of heat which can only determine how much the fermenter
is heated by how high or low it is placed on the fermenting vessel. The lower the position the
more the belt will heat the vessel, the higher it’s placed a smaller amount of heat will transfer
through to the fermenter.
There is very limited control with a heat belt unless you use a dedicated temperature
controller with the belt plugged into this. In this method you get much greater control.
Heating Mats or Pads
Heating mats or pads are devices you place the fermenter on which then transfer heat into the
fermenter.
Heatings mats work in a very similar fashion as heating belts. Most don’t have built in
thermostats or controls so again there is not much precision in their use unless you use a seperate
temperature controller.
Temperature Controlled Refrigerator
Again, the most efficient method of controlling your fermentation temperature is a refrigerator.
Of course, using a refrigerator to warm a fermentation doesn’t make sense.
In the case of heating a fermenter we need to place a heat source in the fridge connected to a
temperature controller. Heat sources could be a storage heater, lightbulb or even a heat mat. The
refrigerator acts simply as a insulated box which keeps the heat in.
The added benefit of a refrigerator fermentation chamber is you can easily heat and cool at
various points of fermentation and then cold crash the fermenter when you want to get the brew
ready for packaging.
FOAM PRODUCTION AND FOAM CONTROL
How is foam generated
The pharmaceutical Industry uses fermenters for production and involves stirring and mixing of
air and gas for operation. However, these natural organisms create unwanted bi-products which
support foam stabilization making it easy for stable foam production.
There are the chances of liquid completely turning into foam if not controlled on time. Anti-
foams are generally used to protect from foam generation, but it leads to unwanted side effects
which is difficult to control.
Overall, anti-foam usage can alter the end results significantly, therefore, measuring and control
of foam generation using foam detectors is necessary.
Precise foam control is important in pharmaceutical industry during fermentation process. The
industry deals with health sector, therefore, accurate result during the process used in the
production of medicines, antibiotics, vaccines and many other drugs is critical.
Foam-Control
 The level of foam in the vessel must be minimized to avoid contamination, this is an
important aspect of the fermentor.
 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.