Professional Documents
Culture Documents
AND CONTROL
A well-stirred tank, one of the most
commonly employed bioreactors,
approximates reasonably well to the
idealized state of perfect mixing. It ensures
a very short mixing time and a high gas
liquid mass transfer on a small scale. This
particular culture conditions are mostly
achieved in so-called high performance
bioreactors.
Introduction
This is very important in microbial
physiological studies under conditions
closely approaching those found in
industrial reactors to be able to distinguish
between the dynamics of cell reactions
and the dynamics of the reaction vessel. In
such bioreactors cells are subjected to an
unchanging environment when circulated
throughout the liquid medium.
A standard stirred tank bioreactor is normally
supplied with a means of measuring temperature,
agitator speed, pH, the incoming air-flow rate and
dissolved oxygen concentration. This basic
instrumentation can be complemented with several
other sensors capable of determining the pressure
drop, dissolved CO2 concentration, biomass, off-gas
analysis, foam control, power and torque, and liquid
flow feeding.
Temperature
Thermocouples are the most widely used
temperatures sensors since they are cheap,
provide rapid response and may be used
to measure both high and low
temperatures.
They are less accurate (0.5 C to 2.2C)
than other sensors, however, and must be
calibrated periodically.
Common instruments for
process automation
General view of the laboratory of the research group
Bioprocesses Control Laboratory with instrumentation
Gas Flowrate
Differential Pressure Producers are
frequently found in industrial applications
given their reliability and easy of use. Their
accuracy ranges from 0.8 % to 5%.
The basic principle is that any obstruction in
the fluid stream generates a pressure loss
that is related to the flowrate of the stream.
The main limitation of such devices is that
the cost in energy from the loss of pressure
cost can be considerable.
In-gas converter (air and oxygen), prototype developed
by Bioprocesses Control Laboratory group ICT Prague
utilises controllers F-201C-FZ (Bronkhorst Hi-Tec,
Netherlands)
Another commonly found air velocity
measurement is the turbine anemometer.
Here, a rotating device is placed in the
path of the fluid, where its rotational speed
is proportional t the fluid velocity and
provides accurate flow measurement over
wide ranges.
Liquid Flowrate
There are many ways of measuring liquid
flowrates ranging from those based on
simple hydraulic properties to those based
on radioactive effects.
Instruments are usually sensitive to noise,
however, interference from other electric
devices must be controlled.
Turbine meters are small electric turbines
that generate an electromotive force
which is proportional to the mean fluid
velocity.
This instruments are reliable, precise and
can be used with clean liquids.
More suited for the dirty liquids sensors are
magnetic sensors, since they are non-
invasive, but which are very expensive.
The instrument generates a magnetic field
that is perpendicular to the liquid flow.
Less expensive Doppler sensors, can also
be used to measure flow rates of dirty
liquids.
Here, a continuous ultrasonic wave (0.5 to
10 MHz) emitted by the instrument is
reflected by the bubbles and suspended
solids in the liquid stream.
Radioactive sensors are based
on the tracking of radioactive
trace species.
These are very precise
instruments and applicable to
any kind of fluid, but they are
very expensive and dealing with
radioactive species is difficult.
Off-Gas Analysis
Analysis of exhaust gases provides
information on the physiological state and
respiration rate of the culture.
On-line measurement of CO2 and O2 can
be performed by gas chromatography
(GC) or special purpose gas analyzers.
The main advantages of GC are that many
compounds can be monitored with the
same instrument over a wider range of
values
Gas Analyzers, on the other hand, are
more precise and provide a faster response
(few seconds). Several such instruments
exist. Paramagnetic analyzers, available for
CO2 and O2, are very precise, do not
require periodic calibration, present low
interference to other gases and have long
lifetimes, but they are expensive.
Infrared instruments , are available for CO2
only, are precise and having a long
lifetime, though they are expensive and
need occasional calibration.
Electrochemical analyzers, only
for O2, are low cost instruments
that provide good precision,
however a fuel cell must be
changed periodically (between
months and 2 years)
Infrared analyzer CO
2
Servomex 1440B
(Servomex International Ltd., Great
Britain) for off-gas analysis
Paramagnetic analyzer CO
2
Servomex 1100A (Servomex International Ltd.,
Great Britain) for off-gas analysis
Analyzer of medium composition HPLC
chromatograph with detectors Waters
UV/VIS 2487 and Waters RI 2410 with two
pumps Waters 515 (Waters Corp.,
Massachusetts, USA)
pH
The pH sensitive electrode (usually made of
glass) contains a buffer solution at a constant
pH. The glass behaves as a membrane that
separates the sample from the buffer solution;
a potential proportional to the pH difference is
generated.
If pH control is critical to bioreactor operation,
measurement redundancy is recommended.
Using three sensors would be wise because
when one gives a very different reading from
the other two it means that it needs cleaning.
Ion Sensitive Field-Effect Transistor - ISFET
Dissolved Oxygen
When controlling dissolved oxygen in a
bioreactor, it is often sufficient to keep the
dissolved oxygen above a certain level.
pO2 (partial pressure of dissolved oxygen)
One of the most specific aspects of the
fermentation monitoring is
pO2 measurement and control.
pO2 control is characteristic only for
fermentation processes. There are different
pO2 control principles:
pO2 is commonly adjusted in % from the
fixed one. The adjusted pO2 value has a
lower and upper limit. The difference
between both these limits is usually 10% -
20%.
Important parameters in pO2 control are
the control limits of the mixer's rotational
speed n: nmin and nmax. It means that,
when controlling pO2, n will vary only within
this range. These limits are determined in
connection with eliminating of different
undesirable phenomena:
Foam
The appearance of foam is a very undesirable
phenomenon, since, in the course of its
appearance, there is a risk to loose an essential part
of the fermentation broth. During the foaming, it is
not possible to perform high-quality analyses and
measurements. For elimination of foam, 2 methods
or their combinations are commonly used:
Additional metering of an antifoam, based on the
information provided by the foam sensor. The given
impulses are relatively low, with long pauses and a
limited metering time. This additional control is
necessary to avoid the possible overdose, since, in
this case, the mass exchange parameters can
decrease dramatically.
Mechanical metering of foam. For this
purpose, an upper drive with a special disk-
type or other type of the mechanical foam
breaking mixer is installed in the
bioreactor's upper cover. If an intensive
foaming begins, then the mechanical
breaking of foam will not help any more.
An optimal solution is the combination of
both the parameters. The application of
Variant 1 is more widely used in laboratory
bioreactors.
Thank you!