Advanced

Chemical Reaction
Engineering
Topic 2d
Bioreactors and Biosynthesis
-Various Bioreactor Designs
 L10-2

Topic Learning Outcomes

After completing this topic, students will be able to:
• Discuss the pseudo-steady-state-hypothesis and
explain how it can be used to solve reaction
engineering problems.
• Write reaction pathways for complex reactions.
• Explain what an enzyme is and how it acts as a
catalyst.
• Describe the Michaelis-Menten enzyme kinetics and
rate law along with its temperature dependence.
• Discuss how to distinguish the different types of
enzyme inhibition.
• Discuss the stages of cell growth and the rate laws
used to describe growth.
• Write material balances on cells, substrates, and
products in bioreactors to size chemostats

Slides courtesy of Prof M L Kraft, Chemical & Biomolecular Engr Dept, University of Illinois, Urbana-Champaign.
 Microbial Kinetics and Bioreactors
• Introduction to Process Biotechnology
– Advantages of Bioprocesses
– New Developments
– Overview of a Bioprocess
• Microbial Kinetics and Bioreactors
– Phases of Growth
– Growth Kinetics
– Batch Bioreactor
– Continuous Bioreactor: Growth and Substrate Uptake
– Inhibition Kinetics
– Product Formation kinetics
• Various Bioreactor Designs and Applications
3
Various Bioreactor Designs

4
Fermenter
View looking down into a 125m3 stainless
steel fermentor
 Industrial Bioreactor

Glacial Lakes Energy in Watertown, South Dakota

47+ million gallon per year ethanol production .

World's Largest Industrial Fermenter (Chem. Eng. News,10-Apr-78)

The fermenter is 200' high and 25 ft diam.

 Broad classification of Bioreactors

• Solid state: water content: 40~ 80%, mostly mold

fermentation on agriculture products and food:
rice, wheat, barley, corn and soybean.
e.g. Rotary drum fermentor
• Submerged systems: water content > 95%
e.g. bacteria, yeast, mold, plant, animal cells
 Overview of bioreactors for
submerged system
operation modes:
- batch: stirred tank
- continuous: chemostat, fluidized-bed
- modified types of the above modes:
fed-batch, chemostat with recycle,
multi-stage continuous reactors

Oxygen supply: Form of biocatalyst:

- aerobic - free cell or enzyme
- anaerobic - immobilized cell or enzyme,
packed-bed, membrane reactor
 Requirements for Cultivation Methods

• Biomass concentration which must remain high

• Sterile conditions being maintained
• Effective agitation so that the distribution of
substances in the reactor is uniform
• Heat removal
• Creation of the correct shear conditions - high
shear may damage cells, low shear may lead to
flocculation or growth on wall and stirrer
 Types of Bioreactors
• Bioreactors vary in size and complexity from a 10 ml volume
in a test tube to computer controlled fermenters with liquid
volumes greater than 100 m3. Their cost varies from a few
cents to a few million dollars.
• In the following sections we will compare the following
reactors
 Solid substrate fermentations
 Shake flasks
 Stirred tank reactors
 Bubble column and airlift reactors
 Fluidized bed reactors
 Standing cultures
• Aerobic solid substrate fermentations are another example
of standing cultures. In these fermentations, the biomass is
grown on solid biodegradable substrates such as water
softened bran, rice or barley.
• The solids may be continuously or periodically turned over to
improve aeration and to regulate the culture temperature.
• One example of a commercial scale, solid substrate
fermentation is the production of koji by Aspergillus oryzae on
soya beans which is part of the soya sauce process.
• Another is mushroom cultivation. Considerable research is
currently being invested into the feasibility of producing
biochemicals by solid substrate fermentations.
Aspergillus niger mycelia
Shake flasks
 Shake flasks
• Shake flasks are commonly used for small scale aerobic
cell cultivation.
• Through continuous shaking of the culture fluid, higher
oxygen transfer rates can be achieved as compared to
standing cultures.
• Shaking continually breaks the liquid surface and thus
provides a greater surface area for oxygen transfer.
• Increased rates of oxygen transfer are also achieved by
entrainment of oxygen bubbles at the surface of the liquid.
 Stirred Tank bioreactors
• For liquid volumes greater than 3 litres, air sparging is
required for effective oxygen transfer.
• The introduction of bubbles into the culture fluid by
sparging, leads to a dramatic increase in the oxygen
transfer area.
• Agitation is used to break up bubbles and thus further
increase kLa.
• Sparged fermenters required significantly lower agitation
speeds for aeration efficiencies comparable to those
achieved in non-sparged fermenters.
• Air-sparged fermenters can have liquid volumes greater
than 500,000 litres.
 Bubble driven bioreactors
• Sparging without mechanical agitation can also be used for
aeration and agitation. Two classes of bubble driven
bioreactors are bubble column fermenters and airlift
fermenters.
• Bubble driven bioreactors are commonly used in the culture
of shear sensitive organisms such as moulds and plant cells.
• An airlift fermenter differs from bubble column bioreactors by
the presence of a draft tube which provides better mass and
heat transfer efficiencies.
• Airlift fermenters are however considerably more expensive
to construct than bubble column reactors.
• There are several designs for air-lift fermenters although the
most commonly used design is one with a central draft tube.
Bubble driven bioreactors
 Airlift bioreactors
• An airlift fermenter differs from bubble column bioreactors
by the presence of a draft tube.
• The main functions of the draft tube are to:
 Increase mixing through the reactor
The presence of the draft tube enhances axial mixing
throughout the whole reactor
 Reduce bubble coalescence.
This presumably occurs due to circulatory effect that
the draft tube induces in the reactor. The circulation
occurs in one direction and hence the bubbles also
travel in one direction.
 Airlift bioreactors
Small bubbles lead to an increased surface area for oxygen transfer.
 Airlift bioreactors
• Equalise shear forces throughout the reactor.
Major reason why the productivity of cells grown in airlift
bioreactors have higher productivities than those grown
in stirred tank reactors.
 Airlift bioreactors
• The major disadvantages of air-lift fermenters are

- high energy requirements

- excessive foaming

- cell damage due to bubble bursting;

particularly with animal cell culture
 Airlift bioreactor
Air-riser and down-comer
• An air-lift reactor is divided into three regions:

- the air-riser
- down-comer
- disengagement zone.
Airlift bioreactor
 Airlift bioreactor
• The region into which bubbles are sparged is called the air-
riser. The air-riser may be on the inside or the outside of the
draft-tube. The latter design is preferred for large scale
fermenters as it provides better heat transfer efficiencies.
• The rising bubbles in the air-riser cause the liquid to flow in a
vertical direction. To counteract these upward forces, liquid will
flow in a downward direction in the down-comer. This leads to
liquid circulation and thus improved mixing efficiencies as
compared to bubble columns.
• The enhanced liquid circulation also causes bubbles to move in
a uniform direction at a relatively uniform velocity. This bubble
flow pattern reduces bubble coalescence and thus results in
higher kLa values as compared to bubble column reactors.
 Airlift bioreactors -
Disengagement zone
 Airlift bioreactors -
Disengagement zone
• The sudden widening at the top of the reactor slows the
bubble velocity and thus disengages the bubbles from the
liquid flow.
• Carbon-dioxide rich bubbles are thus prevented from entering
the downcomer.
• The reduced bubble velocity in the disengagement zone also
leads to a reduction in the loss of medium due aerosol
formation.
• The increase in area will also helps to stretch bubbles in
foams, causing the bubbles to burst. The axial flow circulation
caused by the draft tube also helps to reduce foaming
 Packed bed and trickle flow
bioreactors
• In packed bed bioreactors, immobilized enzymes or
cells are packed in a column.
 Packed bed bioreactors
• The rate of mass transfer between the cells and the medium
depends on the flow rate and on the thickness of the biomass
film on or near the surface of the solid particles.
• Packed bed reactors often suffer from problems caused by
poor mass transfer rates and clogging. Despite this, they are
used commercially with enzyme catalysts and with slowly or
non-growing cells.
• They are also used in the anaerobic treatment of high strength
wastewaters (eg. food processing wastes).
• Large plastic blocks are used as solid supports for the cells.
These blocks have a large surface area for cell immobilization
and when packed in the reactor are difficult to clog.
Fluidised bed reactors
 Fluidized bed reactors
• Fluidized bed bioreactors are one method of maintaining high
biomass concentrations and at the same time good mass
transfer rates in continuous cultures.
• Fluidized bed bioreactors are an example of reactors in which
mixing is assisted by the action of a pump. In a fluidized bed
reactor, cells or enzymes are immobilized in and/or on the
surface of light particles.
• A pump located at the base of the tank causes the
immobilized catalysts to move with the fluid. The pump
pushes the fluid and the particles in a vertical direction.
• The upward force of the pump is balanced by the downward
movement of the particles due to gravity. This results in good
circulation.
 Fluidised bed reactors
• For aerobic microbial systems, sparging is used to improve
oxygen transfer rates.
• A draft tube may be used to improve circulation and
oxygen transfer.
• Both aerobic and anaerobic fluidised bed bioreactors have
been developed for use in waste treatment.
• Fluidized beds can also be used with microcarrier beads
used in attached animal cell culture.
• Fluidized-bed micro carrier cultures can be operated both
in batch and continuous mode.
• In the former the fermentation fluid is recycled in a pump-
around loop.
Fluidised bed reactors
TABLE 1. Basic Bioreactor Design Criteria
___________________________________________________________________
 Microbiological and Biochemical Characteristics of the Cell
System (Microbial, Mammalian, Plant)
 Hydrodynamic Characteristics of the bioreactor
 Mass and Heat Transfer Characteristics of the Bioreactor
 Kinetics of the Cell Growth and Product Formation
 Genetic Stability Characteristics of the Cell System
 Aseptic Equipment Design
 Control of Bioreactor Environment (both macro-and micro-
environment)
 Implications of Bioreactor Design on Downstream Products
Separation
 Capital and Operating Costs of the Bioreactor
 Potential for Bioreactor Scale-up
______________________________________________________________________
TABLE 2. Summary of Bioreactor Systems
__________________________________________________________
Bioreactor Cell Systems Products
Design used
__________________________________________________________
 Air-Lift Bioreactor Bacteria, Yeast and SCP, Enzymes,Secondary
other fungi metabolites, Surfactants

 Fluidized-Bed Immobilized bacteria, Ethanol, Secondary

Bioreactor yeast and other fungi, metabolites, Wastewater
Activated sludge treatment

 Microcarrier Immobilized (anchored) Interferons, Growth factors,

Bioreactor mammalian cells on Blood factors, Monoclonal
solid particles antibodies, Vaccines, Proteases,
Hormones

 Surface Tissue mammalian, tissue Interferons, Growth factors,

Propagator growth on solid surface, Blood factors,
tissue engineering Monoclonal antibodies,
Vaccines, Proteases, Hormones

__________________________________________________________
TABLE 2. Summary of Bioreactor Systems
(Cont’d)
____________________________________________________________________________________________________
Bioreactor Cell Systems used Products
Design
________________________________________________________________________________________

 Membrane Bioreactors, Bacteria, Yeasts, Ethanol, Monoclonal anti-

Hollow fibers and Mammalian cells, Plant bodies, Interferons, Growth
membranes used, cells factors, Medicinal products
Rotorfermentor

 Modified Stirred Immobilized Bacteria, Ethanol, Monoclonal anti-

Tank Bioreactor Yeast, Plant cells bodies, Interferons, Growth
factors

 Modified Packed- Immobilized Bacteria, Ethanol, Enzymes, Medicinal

Bed Bioreactor Yeasts and other fungi products

 Tower and Loop Bacteria, Yeasts Single Cell Protein (SCP)

Bioreactors

________________________________________________________________________________________
TABLE 2. Summary of Bioreactor Systems
(Cont’d)
______________________________________
Bioreactor Cell System used Products
design
_______________________________________________________________________________
 Vacuum Bioreactors Bacteria, Yeasts, Fungi Ethanol, Volatile
products

 Cyclone Bioreactors Bacteria, Yeasts, Fungi Commodity products,

SCP

 Photochemical Photosynthetic bacteria, SCP, Algae, Medicinal

Bioreactors Algae, Cyano bacteria, plant products,
Plant Cell culture, r-DNA Monoclonal antibodies,
plant cells Vaccines, Interferons

________________________________________________________________________________________