TOPIC 7:

INSTRUMENTATION AND
CONTROL SYSTEM

 Bioreactor for industrial application should have low capital and

operating cost.
However, there is no single system which adequately meets the
needs of all biological systems can be constructed.

 Laboratory scale bioreactors liquid volume < 10 L constructed out of Pyrex

glass.
For larger bioreactors, stainless steel is used. Stainless steel = refers to
various alloys of primarily iron, nickel and chromium.
Different grade of SS = 302, 304, 316, 318. (higher the number, the greater the
resilience of the steel.
316 L most widely used (L indicates the steel has low carbon content)
The stainless steel used in bioreactors are polished to a mirror finish (makes
cleaning and sterilization easier).
Components joined in an oxygen-free environment to avoid corrosion, displace
by argon (TIG technique, Total Inert Gas)

STANDARD GEOMETRY MEASUREMENT OF A STIRRED TANK

BIOREACTOR

A mechanically stirred tank bioreactor fitted with a sparger and a

Rushton turbine impeller will typically have relative dimensions (Table 1)

Table 1: Geometry Dimension of Stirred Tank Bioreactor

Typical
values
~0.7-0.8

Remarks

Height of reactor to diameter Ht/Dt

of tank

~1 - 2

European reactors tend to be

taller than those designed in
the USA

Diameter
of
impeller
diameter in tank

1/3 - 1/2

Rushton Turbine reactors are

generally 1/3 of the tank
diameter. Axial flow impellers
are larger.

Ratio
Height of liquid in reactor to HL/Ht
height of reactor

to Da/Dt

Diameter of baffles to diameter Db/Dt

of tank

~0.08 - 0.1

Impeller blade height

diameter of impeller

to W/Da

0.2

Impeller
blade
width
diameter of impeller

to L/Da

0.25

Distance between middle of E/W

impeller blade and impeller
blade height

Depends on the level of

foaming produced during the
fermentation

A tank's height:diameter ratio is often referred to as its

aspect ratio. A stirred tank bioreactor is approximately
cylindrical in shape.
The geometry of the reactor can be defined by the following
ratios:
Dt:Ht

0.50

Da:Dt

0.33

Db:Dt

0.10

A modern mechanically agitated bioreactor:

A BIOREACTOR
An agitator system
An oxygen delivery system
A foam control system
A temperature control
system
A pH control system
Sampling ports
A cleaning and sterilization
system.
A sump and dump line for
emptying of the reactor.

AGITATION SYSTEM (Agitator and Baffles)

Function:
provide good mixing and thus
increase mass transfer rates
through the bulk liquid and
bubble boundary layers.
provide the appropriate shear
conditions required for the
breaking up of bubbles.

Stirrer's shaft seal

- subject to high mechanical loads
- Important for good operation; to avoid risk of contamination; shaft
jamming, possible leak.
- All component with contact to the product must have smooth surfaces and
may not have any recesses
- < 1 L bioreactor = plastic coated magnetic rods are used located on
bearings on the bottom (associated with possible aeration problems) or
suspended for rotational movement.
Stirred vessels > 10 L volume, mechanical drive coupling together with rotating
mechanical seals.
Seal components: carbon and ceramic.
Laboratory scale: simple rotating mechanical seal.
Larger scale: double-action rotating mechanical seals; achieving longer
operation times.
The rotating mechanical seal must be free from cracks and the cavities
between the packing must be steamable.

Rotating mechanical seals

Driver Configuration (the drive for the agitator shaft)

Can be installed either above
through the reactor cover or
from below through the bottom
flange.
Bottom drive; - leaves cover
free for the installation of other
components and connection,
the agitator shaft can be kept
shorter.
Top drive; - easily protected
against leakages; sterility is
easier to maintain.

 Multi-blade disc impellers- produce radial flow and high energy

dissipation density in the proximity of the agitator.
Propeller impellers- create an axial flow (for mammalian cell
cultivation), low shear forces in connection with low oxygen
requirements are applicable
INTERMIG impellers- create both radial and axial flows, for high
viscous media or products
Turbine normally for loop systems(mixing effect is achieved by
circulating the liquid)
EKATO INTERPROP improved heat and mass transfer, high
dispersion of drops and bubbles, homogenous distribution of solid
particles as well as low investment costs.
The function(s) of the agitation system include;
increasing mass transfer rates, especially KLa, through the bulk
liquid
providing appropriate levels of shear
increasing heat transfer rates
reducing the size of boundary layers

Effect of Impeller Spacing on Flow Pattern and Power Absorption

a) Spaced too closely multiple impellers tend to behave like a single large
impeller.
b) Spaced too far apart appearing regions in the liquid (Stagnant area).
- Optimal spacing is about one impeller apart, with the lowest impeller about
one impeller above the bottom of the vessel.

Impellers have 2 distinct functions:

I) To provide mixing by pumping liquid round the vessel. For this function,
it requires a large diameter, low speed impeller with a small number of
blades.
II) To dispersed the injected gas steam as small bubbles and re-disperse
coalesced bubbles. It requires a high speed, small diameter impeller with a
large number of blades
Agitators characteristics:
Radial Flow
- the liquid flow from the impeller is initially directed towards the wall of the
reactor
Axial Flow
- The liquid flow from the impeller is directed downwards towards the base of
the reactor

Radial flow impellers

- contain two or more impeller blades which are set at a
vertical pitch

Radial flow impellers - Shear characteristics

Radial flow
impellers
Eddy swirling of a fluid

Flow
pattern
using
radial flow impellers.

Radial flow impellers - Rushton turbine

Generation
of high shear
conditions by
radial flow
impeller

Radial flow impellers - Rushton turbine

Six bladed Rushton

turbine impeller
Mixing is achieved with the
use of baffles.
higher input energy
Mixing is not efficient as axial
flow mixing

Function of Rushton turbine

impeller.

Axial flow impellers

Marine and hydrofoil impellers

Flow pattern of axial impeller

More energy efficient, effective at
lifting solids from the base of the
tank, low shear properties
Use for shear sensitive process
such as crystallization,
precipitation reactions and culture
of animal cells.
Not suitable for bacterial and
fungal aerobic fermentation
(ineffective breaking up bubbles).

Axial flow impellers - Intermig Impeller

- for microbial fermentation

Bottom: has large axial flow section

The tips of impeller contain finger like
extensions, create a turbulence wake
for breaking bubbles.
Overall shear conditions in the
reactor are lower than would be
generated by a radial flow impeller
(Rushton turbine).

Flow pattern created by INTERMIG

impeller

http://www.ekato.com/en/products/ekato-rmt/products/ekato-impellers/intermig/

Advantages:
>For low to medium product viscosities
>Homogeneous dissipation of introduced energy
>High axial flow velocities, therefore increased heat transfer and
reduced scaling at vessel wall
>Low shear mixing

BAFFLES

Unbaffled bioreactor. Note

the presence of a large
vortex. The liquid is
circulating around the
impeller.

Baffled bioreactor. Note the

presence of small bubbles
from gas entrainment and
the absence of a large
vortex.

Formation of eddies by baffles

Baffles break the liquid flow lines causing the formation of

turbulent eddies.

OXYGEN DELIVERY SYSTEM

Consists of:
a compressor
inlet air sterilization system
an air sparger
exit air sterilization system

A compressor
forces the air into the reactor, need sufficient pressure to force the air;
for large reactors, produce air at 250kPa.The air should be dry and oil
free so as to not block the inlet air filter (not to use instrument air)
Air sterilisation system
to prevent contaminating organisms from entering the reactor as well as
to prevent the microorganism in the reactor from contaminating the air.
Common method: filtration
Smaller bioreactor (<5 L) disc-shaped hydrophobic Teflon membranes
housed in a polypropylene housing.
Teflon tough, reusable and does not readily block.

Membrane filters
housed in
polypropylene
cartridges are used.

Membrane filter housed with

Teflon for air sterilization
Large bioreactor (> 1000L) pleated membrane filters housed in
polyproplylene cartridge.
By pleating (folding and pressing) the membrane, it is possible to create a
compact filter with a very large surface area for air filtration. Large scale
membrane filtration is very expensive process.
filtration is not possible; too expensive; Heat sterilisation (steam is use to
sterilize the air)

For small reactor, the exit air system, will include a condenser.

Condenser
simple heat
exchanger
through which
cool water is
passed; minimize
water evaporation
and the loss of
volatiles; Drying
the air also
prevents blocking
the exit air filter
with water

AIR SPARGER
The air sparger breaks the
incoming air into small
bubbles
The sparger must located
below the agitator to
facilitate bubble break up
Ring Sparger

During emptying of a
fermenter, it is important that
the air feed valve is closed.
This will minimize
contamination of the inlet air
line.
Formation of bubbles from ring sparger

Effect of agitation speed on bubbles distribution in liquid

Slow impeller speed
bubbles will not be broken,
rise directly to to the surface,
accumulate and coalesce
under impeller leading to
from large bubbles and low
oxygen transfer rates.

Slow impeller
speed

Fast impeller
speed

Fast speed smaller

bubbles will be generated,
move throughout the reactor
increasing the gas hold up
and bubble residence time.

AIR FLOW RATES

Volume per volume per minute, vvm.

The airflow rate and liquid volume must have the same basal unit. The
air flow rate must be expressed in terms of volume per min.

CONDENSOR TEMPERATURE

A
cold
condenser
temperature can help to
control the foam. The density
of the foam increases when
it moves from the warm
headspace volume to the
cold condenser region. This
causes the foam to collapse.

CONTROL SYSTEMS

-The success of a fermentation depends upon the existence of defined

environmental conditions for biomass and product formation.

-Thus temperature, pH, degree of agitation, oxygen concentration in the
medium and other factors may have to kept constant during the process
-- Require careful monitoring of the fermentation so that any deviation from the

specified optimum might be corrected by a control system

--Monitoring equipment should be capable of being linked to a suitable control

system as well as producing information indicating fermentation progress

It is also important to consider the need for a sensor and its associated control
system to interface with a computer.

Process sensors and their possible control functions

Category

Sensor

Possible control function

Physical

Temperature

Heat/cool

Pressure

Air outlet and inlet

Agitator shaft power

on/off

Agitation speed (rpm)

on/off

Foam

Foam control

Flow rate

Change flow rate

pH
Redox

Acid or alkali addition, carbon source feed

rate
Additives to change redox potential

Oxygen

Change feed rate

Exit-gas analysis

Change feed rate

Medium analysis

Change in medium composition

Chemical

Sensors-

Can be characterised by the indication of its

operation in relation to its application for process control

Off-line sensor not part of the fermentation equipment. The

measured values is not directly available for process control. e.g. of offline monitored variable: Bioreactor volume, flow of acidic and alkaline
titrant, flow of antifoam agent, viscosity; nutrient concentration

On-line sensor Part of the fermentation equipment. e.g. On-line

monitored variable: dissolved oxygen tension, off-gas CO2, O2
concentration;.

In-line sensor Part of the fermentation equipment and directly

used for process control. Ex: air flow, level of foaming, flow nutrient,
stirrer speed, temp, pH.

CONTROL SYSTEMS (Component)

Basic Component of control loop:
1. A measuring element.
-senses a process property such as flow, pressure, temperature, etc., and
generates a corresponding output signal
2. A controller
-The controller compares the measurement signal with a predetermined
desired value (set point) and produces an output signal to counteract any
differences between the two
3. A final control element.
-receives the control signal and adjusts the process by changing a valve
opening or pump speed and causing the controlled process property to
return to the set point.

Manual Control
Simple, e.g.- manual control of a steam valve to regulate the
temperature of water flowing through pipe; manual control for
the addition of antifoam
Depends on the skill of the individual operatives in knowing when and how

much adjustment to make.

Costly in labour

Automatic Control
- Certain modifications are necessary when an automatic control loop is
used.
-The measuring element must generate an output signal which can be
monitored by an instrument
-For temperature control; Eg Thermometer changed to thermocouple, which
is connected to a controller which in turn will produce a signal which will
operate the steam valve
Types:
1. Two-position controllers (On/Off).
2. Proportional controllers
3. Integral controllers.
4. Derivative controllers.
P, I & D - These control methods may be used singly or in combinations in applying
automatic control to a process depending on the complexity of the process

1) Two-Position Controllers (ON/OFF)

the simplest automatic controller
has a final control unit (valve, switch, etc.) which is
either fully open (On) or fully closed (Off).
Example: Temperature control by the thermostat
operating with On/Off
-There will be a delay in response when the
temperature reaches the set point before the heating
source is switched off.
- On the other hand, the water will continue
cooling after the heating source is switched off.

* It is important establish that the maximum and minimum values are

acceptable for the specific process
On/Off control is not satisfactory for controlling any process
parameter where there is likely to be large sudden changes from
the equilibrium. In these cases alternative forms of automatic
control must be used.

2) Proportional Control
The change in output of the controller is proportional to the
input signal produced by the environment change which has
been detected by a sensor

M = Mo + Kc
Where,
M = output signal,
Mo = controller output signal when there is no error,
Kc = controller gain or sensitivity,
= the error signal.
Hence, greater the error (environmental change) the larger is the
initial corrective action which will be applied

3) Integral Control
The output signal of an integral controller is determined by the
integral of the error input over the time of operation. Thus:
M = Mo + 1/ Ti dt
where ,Ti = integral time.
The controller output signal changes relatively slowly at first as
time is required for the controller action to integrate the error.

4) Derivative Control
- The controller senses the rate change of the error signal
and contributes a component of the output signal that is
proportional to a derivative of the error signal . Thus:
M = Mo + Td [d/dt]
where ,Td = a time rate constant.

Combinations of methods of control

Three combinations of control systems are used in
practice:
(a) Proportional plus integral.
(b) Proportional plus derivative.
(c) Proportional plus integral plus derivative.

TYPES OF CONTROLLER
i) AGITATOR SHAFT POWER (to measure the power
consumption)
a watt meter (sensor) attached to the agitator motor
will give a fairly good indication of power uptake
Torsion dynamometers can be used in small-scale
applications.
strain gauges mounted on the shaft within the
fermenter are the most accurate method of
measurement and overcome frictional problems

ii) RATE OF STIRRING

important to monitor the rate of rotation (rpm) of the stirrer shaft
tachometer is used for this purpose; may employ electromagnetic induction,
voltage generation, light sensing or magnetic force as detection mechanisms
iii) FLOW MEASUREMENT AND CONTROL
One of the simplest methods for measuring gas flow to a fermenter is by
means of a variable area meter.
Rotameter, (vertically mounted glass tube with an increasing bore and
enclosing a free-moving float which may be a ball or a hollow thimble )
should not be sterilized and are therefore normally placed between a gas
inlet and a sterile filter

iv) TEMPERATURE CONTROL SYSTEM

The temperature in a vessel or pipe is the most important parameter to
monitor and control in any process
Use of water jackets or pipe coils within a fermenter
The temperature control system consist of; - temperature probe; heat
transfer system

Temperature Probe/Electrode
Mercury-in-glass-thermometers used directly in small bench
fermenters, fragile, use of indication not for automatic control.
Bimetalic thermometers- consists of a bimetallic helical coil
surrounded by a protecting tube or well
Pressure bulb thermometers - a pressure gauge connected by smallbore tubing, which may be up to 60 m in length, to the detecting bulb
(12 X 125 mm). The whole system is gastight and filled with an
appropriate gas or liquid under pres- sure (2800-8000 kN m-2).
Thermocouples - normally operated at ambient temperatures
Electrical resistance thermometers - contains the resistance element, a
mica framework (for very accurate measurement) or a ceramic
framework (robust but for less accurate measurement) around which
the sensing element is wound
Thermistor- semiconductors made from specific mixtures of pure
oxides of iron, nickel and other metals.

Heat Transfers System

The jacket will typically be
"dimpled" to encourage turbulence
in the jacket and thus increase the
heat transfer efficiency
. Coils have a much higher heat
transfer efficiency than jackets.
However coils take up valuable
reactor volume and can be difficult
to clean and sterilize (lead to
contamination); poor mixing

Jacketed vessel for temperature

control

Laboratory scale
reactors

Pilot and
production scale
reactors

Heating requirements

Electric heaters

Cooling requirements

Tap water or
refrigerated water
baths

Steam generated in
boilers
Cooling water
produced by cooling
towers or refrigerants
such as ammonia.

v) pH CONTROL SYSTEM
The pH probe is typically steam sterilizable . The electrodes may be
silver/silver chloride with potassium chloride as an electrolyte.
The pH control system consists of:
A pH probe
Control units and neutralizing agents

pH control system - Setpoint and deadband

Deadband: to prevent excessive alkali and acid addition

If the deadband is too small,

then it is possible that pH
will often overshoot and
undershoot the deadbands
leading to excessive alkali
and acid addition. The trade
off is that a wide deadband
will lead to less precise pH
control

vi) MEASUREMENT AND CONTROL OF DISSOLVED

OXYGEN
Most aerobic fermentations it is essential to ensure that the
dissolved oxygen concentration does not fall below a
specified minimal level
In small fermenters (1 L), the commonest are galvanic and
have a lead anode, silver cathode and employ potassium
hydroxide, chloride, bicarbonate or acetate as an electrolyte
Galvanic - Because of the relatively slow movement of oxygen
across the membrane, this type of electrode has a slow response
of the order of 60 seconds to achieve a 90% reading of true value.
These electrodes are therefore suitable for monitoring very slow
changes in oxygen concentration and are normally chosen
because of the compact size and relatively low cost

Construction of dissolved-oxygen electrode:

(a) galvanic,
(b) polarographic.

Polarographic electrodes - commonly used in pilot and production

fermenters, needing instrument ports of 19 or 25 mm diameter ; Response
times of 0.05 to 15 seconds to achieve a 95% reading

Control Units and Methods of DO control

The control of dissolved oxygen (DO) concentration in the
culture could be either by ;
1. variation of the air flow rate
2. variation of the agitation speed (the rpm of the
impeller) or combination of both processes.
3. gas blending system

vii) FOAM CONTROL SYSTEM

Foam control is an essential element of the operation of a
sparged bioreactor.

Excessive foam formation can lead to

blocked air exit filters and to pressure
build up in the reactor. The latter can
lead to a loss of medium, damage to the
reactor and even injury to operating
personnel

Factors affecting antifoam requirements

The following factors affect the foam formation
and the requirement of antifoam addition:

the fermentation medium

products produced during the fermentation
the aeration rate and stirrer speed.
the use of mechanical foam breakers
the head space volume
condenser temperature

Formation of foaming in culture

>> Media rich in proteins will tend to foam more readily than
simple media

Aeration rate and stirrer speed

Higher stirrer speeds and higher aeration rates increase
foaming problems

Fast stirring speed

Slower stirring speed

Mechanical Foam Breakers

Mechanical foam breakers can eliminate or at least reduce the antifoam
requirement. These devices generate sit above the liquid and generate high
shear forces which break the bubbles in the foam. High shear agitators and
nozzles connected to high shear pumps are often used. The foam is sucked
into a high shear device and in the process is broken up.

The action of mechanical

foam breaker

Headspace volume

The headspace plays

an important role in
foam control
The
larger
the
headspace
volume,
then the greater the
tendency for the foam
to collapse under its
own weight

Antifoam Addition System

When the upper level

probe is below the
foam level, no current
will pass between the
level probes and the
antifoam pump
remains turned off.

When the upper level

probe is immersed in the
foam layer, a current is
carried in the foam. This
causes the antifoam to
turn on.

Foam is typically detected

using two conductivity or
"level" probes. One probe
is immersed in the
fermentation liquid while
the other placed above the
liquid level. When the
foam reaches the upper
probe, a current is carried
through the foam

Foam sensor and control unit

EXIT GAS ANALYSIS

The measurement and recording of the effluent gas composition is
important in many fermentation studies.
By observing the concentrations of carbon dioxide and oxygen in the entry
and exit gases in the fermenter and knowing the gas flow rate it is possible to
determine the oxygen uptake of the system, the carbon dioxide evolution
rate and the respiration rate of the microbial culture

Carbon dioxide is commonly monitored by infrared

analysis using a positive filtering method
The oxygen concentration can be determined by a
paramagnetic gas analyser