10

i) density of fluid : A float (ball / bucket) float in water, sink in steam. When it floats it
closes and when it sinks it opens the valve
ii) temperature of fluid : It has water / alcohol mixture which senses the change in
temperature. This mixture expands in hot steam and closes the valve. When it
contracts in cool water opens the valve.
iii) kinetic effect of fluid in motion : if a low density steam is flowing it will be high
velocity. Like wise high density will flow with low velocity. The conversion of
pressure energy into kinetic energy control the opening and closing

1.3 TYPES OF FERMENTERS

The main function of a fermenter is to provide a controlled environment for the growth of
microorganisms or animal cells, to obtain a desired product. Few of the bioreactor types are
discussed below:

1.3.1 STIRRED TANK FERMENTER

Microbial fermentations received prominence during 1940's namely for the production of
life saving antibiotics. Stirred tank reactor is the choice for many (more than 70%) though it is
not the best. Stirred tank reactor’s have the following functions: homogenization, suspension of
solids, dispersion of gas-liquid mixtures, aeration of liquid and heat exchange. The Stirred tank
reactor is provided with a baffle and a rotating stirrer is attached either at the top or at the bottom
of the bioreactor. The typical decision variables are: type, size, location and the number of
impellers; sparger size and location. These determine the hydrodynamic pattern in the reactor,
which in turn influence mixing times, mass and heat transfer coefficients, shear rates etc. The
conventional fermentation is carried out in a batch mode. Since stirred tank reactors are
commonly used for batch processes with slight modifications, these reactors are simple in design
and easier to operate. Many of the industrial bioprocesses even today are being carried out in
batch reactors though significant developments have taken place in the recent years in reactor
design, the industry, still prefers stirred tanks because in case of contamination or any other
substandard product formation the loss is minimal. The batch stirred tanks generally suffer due to
their low volumetric productivity. The downtimes are quite large and unsteady state fermentation
imposes stress to the microbial cultures due to nutritional limitations. The fed batch mode
adopted in the recent years eliminates this limitation. The Stirred tank reactor’s offer excellent
mixing and reasonably good mass transfer rates. The cost of operation is lower and the reactors
can be used with a variety of microbial species. Since stirred tank reactor is commonly used in
chemical industry the mixing concepts are well developed. Stirred tank reactor with immobilized
cells is not favored generally due to attrition problems; however by separating the zone of
mixing from the zone of cell culturing one can successfully operate the system.
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Fig.2 Stirred tank fermenter

1.3.2 AIR-LIFT FERMENTER

Airlift fermenter (ALF) is generally classified as pneumatic reactors without any

mechanical stirring arrangements for mixing. The turbulence caused by the fluid flow ensures
adequate mixing of the liquid. The draft tube is provided in the central section of the reactor. The
introduction of the fluid (air/liquid) causes upward motion and results in circulatory flow in the
entire reactor. The air/liquid velocities will be low and hence the energy consumption is also
low. ALFs can be used for both free and immobilized cells. There are very few reports on ALFs
for metabolite production. The advantages of Airlift reactors are the elimination of attrition
effects generally encountered in mechanical agitated reactors. It is ideally suited for aerobic
cultures since oxygen mass transfer coefficient are quite high in comparison to stirred tank
reactors. This is ideal for SCP production from methanol as carbon substrate. This is used mainly
to avoid excess heat produced during mechanical agitation.

Fig.3 Air-lift fermenter

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(a) (b)

Fig.4 (a) Inner loop air lift fermenter (b) Outer loop air lift fermenter

1.3.4 FLUIDISED BED BIOREACTOR

Fluidized bed bioreactors (FBB) have received increased attention in the recent years due
to their advantages over other types of reactors. Most of the FBBs developed for biological
systems involving cells as biocatalysts are three phase systems (solid, liquid & gas). The
fundamentals of three phase fluidization phenomena have been comprehensively covered in
chemical engineering literature. The FBBs are generally operated in co-current upflow with
liquid as continuous phase and other more unusual configurations like the inverse three phase
fluidized bed or gas solid fluidized bed are not of much importance. Usually fluidization is
obtained either by external liquid re-circulation or by gas fed to the reactor. In the case of
immobilized enzymes the usual situation is of two-phase systems involving solid and liquid but
the use of aerobic biocatalyst necessitate introduction of gas (air) as the third phase. A
differentiation between the three phase fluidized bed and the airlift bioreactor would be made on
the basis that the latter have a physical internal arrangement (draft tube), which provides aerating
and non-aerating zones. The circulatory motion of the liquid is induced due to the draft tube.
Basically the particles used in FBBs can be of three different types: (i) inert core on which the
biomass is created by cell attachment. (ii) Porous particles in which the biocatalyst is
entrapped.(iii) Cell aggregates/ flocs (self-immobilization). In comparison to conventional
mechanically stirred reactors, FBBs provide a much lower attrition of solid particles. The
biocatalyst concentration can significantly be higher and washout limitations of free cell systems
can be overcome. In comparison to packed bed reactors FBBs can be operated with smaller size
particles without the drawbacks of clogging, high liquid pressure drop, channeling and bed
compaction. The smaller particle size facilitates higher mass transfer rates and better mixing. The
volumetric productivity attained in FBBs is usually higher than in stirred tank and packed bed
bioreactors. There are several successful examples of using FBBs in bioprocess development.
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Fig.5 Fluidised bed bioreactor

1.3.5 PACKED BED BIOREACTOR

Packed bed or fixed bed bioreactors are commonly used with attached biofilms especially
in wastewater engineering. The use of packed bed reactors gained importance after the potential
of whole cell immobilization technique has been demonstrated. The immobilized biocatalyst is
packed in the column and fed with nutrients either from top or from bottom. One of the
disadvantages of packed beds is the changed flow characteristic due to alterations in the bed
porosity during operation. While working with soft gels like alginates, carragenan etc the bed
compaction which generally occurs during fermentation results in high pressure drop across the
bed. In many cases the bed compaction was so severe that the gel integrity was severely
hampered. In addition channeling may occur due to turbulence in the bed. Though packed beds
belong to the class of plug flow reactors in which backmixing is absent in many of the packed
beds slight amount of backmixing occurs which changes the characteristics of fermentation.
Packed beds arc generally used where substrate inhibition governs the rate of reaction. The
packed bed reactors are widely used with immobilized cells. Several modifications such as
tapered beds to reduce the pressure drop across the length of the reactor, inclined bed, horizontal
bed, rotary horizontal reactors have been tried with limited success.

Fig.6 Packed bed bioreactor

1.3.6 BUBBLE COLUMN FERMENTER

Bubble column fermenter is a simplest type of tower fermenter consisting of a tube which
is air sparged at the base. It is an elongated non-mechanically stirred fermenter with an aspect
ratio of 6:1. This type of fermenter was used for citric acid production.
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Fig.7 Bubble column fermenter

1.4 CONTROL AND MONITORING FERMENTATION SYSTEM

The integral part of a high-quality bioreactor is a process controller. Such a controller is

commonly specially formed for a definite bioreactor brand. This is rather connected with the fact
that microorganism cultivation processes have relatively high requirements in respect to
precision and sophistication. All this is despite the fact that almost all bioreactors monitor and
regulate the same values actually invariably.

There are three types of sensors used in fermenter. They are,

In-line sensors form integral part of fermenter. The directly measured value controls the process.
Eg. Antifoam probe

On-line sensors form integral part of fermenter. The measured value must be entered into
control system to control process. Eg. Ion specific sensors, mass spectrophotometer.
Off-line sensors do not form integral part of fermenter. The measured value must be entered into
control system for data collection.

1.4 TEMPERATURE

Heat is generated from any fermentation process due to microbial activity and agitation.
The heat control in small scale is carried out by thermostatically controlled bath, internal heating
coils, heating jacket(water),silicon heating jacket ;large scale: inter coils and cold water
circulation. Cooling water is required less for bacteria but more for fungi (due to low optimum
temperature for growth).

1.4.1 TEMPERATURE MEASURING DEVICES

The devices are classified into three major types viz. Non-electrical type, Electrical type
and Radiation type.
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Non-electrical Type:

Non-electrical

Bimetallic thermometer Liquid-in-glass thermometer Pressure thermometer

Mercury in steel Constant volume Vapour pressure

Electrical type:

Electrical Type

Electrical resistance Thermo electrical resistance

Metallic resistance Semi conductor resistance

Radiation type:

Radiation type

Total radiation Partial radiation IR pyrometer

Among the various types listed above three types of temperature measuring devices such
as Mercury-in-glass, Electrical resistance and Thermistors are used widely in fermentation
process.

Mercury-in-glass thermometers: Mercury enclosed in bulb expands with increase in

temperature. Expansion is read as measure of temperature and is used only as indication.

Electrical resistance: The property of some metals whose resistance changes with change in
temperature is used to measure temperature. Bulb with mica is used for accurate measurement
and ceramic for less accurate measurement. This is covered by temperature sensing platinum
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wire. Leads from bulb connected to measuring device Wheat-stone bridge circuit. It is
advantageous in showing fast response to change in temperature.

Thermistors: These are semi conductors of pure oxides of iron, nickel or other metals. They
exhibit large change in resistance with a small temperature change and hence highly sensitive
even little temperature change.

1.4.2 TEMPERATURE CONTROLLING DEVICE

Temperature is an important parameter of fermentation, since, in the cultivation of many

microorganisms, the temperature deviation by a couple of degrees can diminish dramatically the
growth and biosynthesis productivity. The cultivation temperature is commonly monitored with
accuracy not less than ± 0.5°C. For temperature measurements, stainless steel Pt100 sensors are
normally used. The temperature in laboratory bioreactors is controlled by one of the following
ways:
1. A heater is located inside the bioreactor vessel, and cooling is ensured by thin-wall
pipes located in the upper cover, which are connected with an electromagnetic valve
with the cooling water.
2. Heating and cooling proceed in a thermostat, and this thermostatted water, with the
help of a pump, circulates through the bioreactor jacket.

Variant 1 is less complicated, and it ensures a more economic constructive solution. This
variant works very well for small bioreactors with the volume up to about 5 litres. Variant 2
ensures a more even distribution of heat throughout the bioreactor volume, which is essential in
microorganisms' cultivation.
In the temperature regulation process, the main reason for the regulation inaccuracy is the
incorrectly chosen PID parameters. This manifests itself as temperature oscillations. To regulate
the temperature precisely, the main obstacle is often the too high minimal portion of the cooling
water. In this connection, the valves in the cooling water supply line should be adjusted
correspondingly. Another factor for the regulation accuracy is the area and density of the heat
transfer surface, since the higher is inertia, the more difficult is to reach a higher accuracy.

Components used for controlling temperature in a fermenter are water inlet, pressure
regulator, magnetic inlet valve, heater, circulation pump, jacket, cooling water valve, cold finger,
pt-100 sensor, controller, exit gas cooler and drain to remove overflow.
 Manual inlet is used to fill the jacket and closed to allow 30 to 50 drops per minute and
compensate evaporation loss.
 Low voltage heating system - when heating is needed. Controlled by controller-regulated
by pt-100 sensor
 Pipe line - goes to jacket and re circulated to point before heating system
 Cooling water valve - controlled by controller-regulated by pt-100 sensor. Enters through
cold finger which is closed pipeline/coil passes through top plate. Act as heat exchanger
with culture
 Exit gas cooler - receives water from point before cooling water valve
 Drain - water from exit gas cooler, overflow from cold finger, overflow from water
jacket(drained in to lowest point)
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 pt-100 - platinum resistance electrode. Indicate vessel/culture temperature by relating

changes in electrical resistance of sensor to temperature
 Novel sparger – impeller design – improves aeration at slow speeds
 Swept back ports – produce negative pressure ideal for animal cell culture

1.4.3 GAS FLOW RATE

1.4.3.1 GAS FLOW RATE MEASURING DEVICE

Flow rate can be measured by simple variable area meter.

a) Rotameter: is a vertically mounted glass tube with an increasing bore size and enclosing a free
moving float (a ball or a hollow thumble). The position of float indicates the flow rate. This is
less accuracy at low flow rate. Since air coming out of it is non-sterilized, it is placed between
inlet and filter. Metal tubes can replace glass tubes. Float position determined by magnetic or
electrical techniques. This can be used to measure gas and liquid flow rates.
b) Thermal mass flowmeter: In this 2 thermistors placed (T1, T2) are placed upstream and
downstream of the heat source which may be inside or outside the piping. The mass flow rate Q
can be calculated from the equation,
H = QCp(T1-T2)
Where, Cp – specific flow rate of the gas, T1- temperature of gas before heat transfer and T2 –
temperature of gas after heat transfer
This change could induce voltage signal which could be used in data logging.

Fig.8 Thermal mass flow meter

As shown on the scheme, the heater R H and sensor R S need to be connected in a bridge
circuit. It is essential to determine the correct values of the resistors R1, R2, and R3. The bridge
is in balance as soon as the desired temperature difference between R S and R H has reached e.g.
10K. At a particular flow velocity the bridge voltage U_Br needs to be controlled in dependence
of the bridge balance V1-V2. The values for R1..R3 are depending on the measuring range, the
temperature difference dT and the medium which should be measured. We will provide you with
the values of R1..R3 , depending on the application. For calibration the R2 needs to be adjusted
within a range of ± 5%. The method of adjustment relies on the application.