Bioreactors and

Fermenters

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Course Objectives
 This lesson will discuss bioreactors including
basic operations, typical instrumentation
configurations and calibration methods
 We will discuss the basic operation of a
Bioreactor/Fermenter
 We will discuss some of the instrumentation
applicable to a client bioreactor

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Bioreactor Basics
 Bioreactors are specialized tanks used for
growing cells
 Bioreactors are used in many different
processes from brewing beverages to
developing pharmaceutical products

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Bioreactor Basics
A bioreactor is a tank that maintains a nutrient
level necessary for cell growth, as well as
providing an environment that meets the
temperature, pH and oxygen levels that are
needed for the process
The steps in the bioreactor process
(fermentation) are as follows:

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Bioreactor Basics
 Batch Fermentation: A sterilized nutrient
solution in the bioreactor is inoculated with
microorganisms, and incubation begins. At
this stage nothing is added to the process
except oxygen, an antifoaming agent, and
acids or bases to control the pH level. The
biomass concentration at this point generally
changes constantly as a result of the
metabolism of the cells

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Bioreactor Basics
 After inoculation of the sterile nutrient solution
with microorganisms, four typical phases of
growth occur
 Lag Phase: During this phase, there is a
physiochemical equilibration between the
microorganism and the environment which
follows the inoculation where there is very
little growth

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Bioreactor Basics
 Log Phase: Towards the end of the lag phase, cells
have adapted to the new conditions of growth. The
growth of the cell mass now approaches a doubling
of cell number per unit of time for bacteria and
yeast, or a doubling of biomass per unit of time for
filamentous organisms such as fungi
 As long as there exists an excess of substrate,
growth rate of the cell mass remains constant during
this phase

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Bioreactor Basics
 Stationary Phase: When the substrate has been
metabolized or toxic substances have formed,
growth rate begins to slow down or completely stop.
The biomass increases gradually, or remains
constant during this phase, however composition of
the cells may change
 Due to lysis (destruction of the cell membrane) new
substrates are released which may serve as energy
sources for the slow growth of survivors.

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Bioreactor Basics
 Death Phase: During the Death Phase, the
energy reserves of the cells are exhausted.
Cells are now dying at an exponential rate
 During most processes, the fermentation is
interrupted somewhere between the log
phase and the death phase

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Bioreactor Basics
 Fed Batch Fermentation: In the fed batch
fermentation process, the substrate is added
in increments as fermentation progresses.
The critical elements of the nutrient solution
are added in small concentrations at the
beginning of the fermentation, and these
substances continue to be added in small
doses during the production phase

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Bioreactor Basics
 Continuous Fermentation: Sterile nutrient
solution is added to the bioreactor
continuously and an equivalent amount of
converted nutrient solution with
microorganisms is simultaneously taken out
of the system

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Bioreactor Basics
 Microbial Fermentation is a three phase system.
 Phase 1: The liquid phase contains dissolved
nutrients, substrates and metabolites.
 Phase 2: The solid phase consists of individual
cells, pellets, insoluble substrates, or precipitated
metabolic products.
 Phase 3: The gaseous phase provides a reservoir
for oxygen supply and CO2 removal.

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Bioreactor Basics
 Stirring and Mixing is crucial for the transfer of
energy, nutrients, substrate and metabolites.
 For the three phases, mixing brings about the
following:
 Dispersion of air in the nutrient solution.
 Homogenization to equalize the temperature and
concentration of nutrients.
 Suspension of microorganisms and solid nutrients.
 Dispersion of immiscible liquids.

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Bioreactor Basics
 One of the most critical factors in the fermentor
operation is provision for adequate gas exchange.
 Oxygen is the most important substrate for microbial
metabolism, and carbon dioxide is the most
important gaseous metabolic product.
 The mass transfer of oxygen to a liquid is
characterized by the oxygen transfer rate or the
volumetric oxygen transfer coefficient. These
values are a critical parameter of the bioreactor
process.

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Bioreactor Basics
 For optimal yield, the fermentation process
must be carried out at constant temperature.
 Finally, sterilization of the cultures at all
stages is crucial to the process.
 Factors that affect the sterilization of the
cultures include the composition of the
culture medium, the pH value, and the size of
the suspended particles.

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Bioreactor Basics
 It is for all of the stages of the
bioreactor/fermentation process mentioned
above, that instruments that monitor
agitation, temperature, pH and DO2
(dissolved oxygen) content are all critical to
the success of the process.
 The next section covers the instrumentation
involved in a typical Bioreactor.

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Bioreactor Instrumentation
The exact instrumentation configuration is
different from one bioreactor to the next, but
all bioreactors contain a similar set of
instrumentation.
Bioreactors contain a number of temperature
measuring devices measuring many
parameters including the following (Actual
usage vary from one bioreactor to another):

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Bioreactor Instrumentation
 Tank Temperature
 Jacket Temperature
 Inoculums Temperature
 Media Temperature
 Titrant Temperature
 Sample Temperature
 Harvest Temperature
 CIP Steam Temperature
 Agitator Steam Temperature

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Bioreactor Instrumentation
Bioreactors contain analytical measurement
instruments including one or more pH transmitters
and one or more DO2 transmitters.
Bioreactors contain air flow measurement instruments
measuring Sparge Gas, O2, CO2 and Headspace
Air flow into the tank.
Bioreactors have a number of pressure measuring
devices including pressure gauges and pressure
transmitters measuring many parameters including
the following:

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Bioreactor Instrumentation
 Tank Pressure
 Media Pressure
 Steam Pressure
 Compressed Air Pressure

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Bioreactor Instrumentation
Bioreactors typically have a level sensing
device of some type.
Bioreactors have a speed sensing instrument
attached to the agitator motor to monitor the
amount of agitation.

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Summary
 This lesson covered the basics of the
fermentation process, and the basic
functioning of a bioreactor including a
discussion of the critical parameters.
 The instrumentation used in a typical
bioreactor system were discussed.

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Where to Get More
Information
 There are many possible sources of
information about this subject including white
papers on the fermentation process,
discussion with scientists and engineers at
client sites, and Manuals and Standard
Operating Procedures for the actual
Bioreactor system being worked on.

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