BTK5301 (Bioreactor System)

22/07/10

BIOREACTOR
DESIGN
Prof Madya Dr. Umi Kalsom Md Shah
Dept. Bioprocess Technology
 Types of Bioreactor
Major Function:
To provide controlled environment for
growth of microorganism (or mixture) to
obtain desired product.
Points to be considered in designing and
constructing bioreactor:
1)               Microbiological and biochemical characteristics
of cell systems.
2)               Hydrodynamic characteristics of the bioreactor.
3)               Mass and heat characteristics of the bioreactor.
4)               Kinetic of cell growth and product formation.
5)               Genetic stability characteristics of the cell system.
6)               Aseptic equipment design.
7)               Control of bioreactor environment (macro and
microenvironments).
8)               Implication of bioreactor design on downstream
product separation.
9)               Capital and operating costs of the bioreactor.
10)         Potential for bioreactor scale up.
Vary in size and complexity,
test tube (10 ml) to
computer controlled
fermenters (>100 m3).
1. STANDING CULTURES

Little or no power used for aeration.

Aeration - dependent on transfer of oxygen
through the still surface of the culture.

Poor Rate of oxygen transfer - due to small

surface area.

Used in small-scale (oxygen supply is not

critical).

Ex: biochemical tests for identification of

bacteria (test-tubes containing 5-10 mL of
media).
 a. T- Flasks
Small scale culture of animal cells. Incubated
horizontally to increase the surface area for
oxygen transfer.

Surface aeration rate can be increased

using large volume flasks.
b. Fernback Flasks

Fig. 1 shows a 3 L "Fernback" flask

containing 1 L of medium and a 250 mL
Erlenmeyer flask containing 100 mL of
medium. Note how the former has a large
surface area.

Figure 1
Fernback
flask
Large Pyrex flasks are used for small-
scale production of fermented products.
Ex: Kombucha tea, (a tea brewed by
mixture of yeasts and acetic acid
bacteria).
c. Surface Cultures

Standing culture aeration is not

restricted to the laboratory. In some
countries, where availability of
electricity is unreliable, citric acid is
produced using surface culture
techniques.
Aspergillus niger mycelia are grown on
surface of liquid media in large shallow
trays. The medium is neither gassed
nor agitated.
 Ex: Aerobic solid substrate
fermentations (biomass is grown
on solid biodegradable substrates
such as water softened bran, rice or
barley, solids continuously or
periodically turned over to improve
aeration and to regulate culture
temperature).
Ex: 1. Production of koji by
Aspergillus oryzae on soya beans,
(part of soya sauce process).
2. Mushroom cultivation.
 2. SHAKE FLASKS

For small scale cell cultivation

(continuous shaking of culture fluid),
higher oxygen transfer rates.
 
Shaking breaks liquid surface and
provides 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.
 
 a. Factors Affecting KLa (volumetric
oxygen transfer rate)
 
Rate of oxygen transfer is dependent on:
1.  shaking speed,
2. liquid volume,
3. shake flask design

KLa increases with

KLa decreases with KLa is higher when
liquid surface area
liquid volume
baffles are present
     KLa increase with shaking speed.
 
   At high shaking speeds, bubbles
become entrained into medium to
further increase oxygen transfer rate.
 
2. Baffles
     Presence of baffles in flasks will
increase oxygen transfer efficiency,
particularly for orbital shakers.
    High level of foam formation in the
baffled flask is due to higher level of gas
entrainment.

Unbaffled flask Baffled flask

3. MECHANICALLY STIRRED
BIOREACTORS

Most important bioreactor for industrial

application, (pharmaceutical industry).
  Low capital cost, low operating costs,
best understood and flexible.
  Non-sparged mechanically agitated
bioreactors can supply sufficient aeration
for microbial fermentations with liquid
volumes up to 3 L.
              However, stirring speeds of up to
600 rpm may be required before the
culture is not oxygen limited.

In non-sparged reactors, oxygen is

transferred from head-space above
fermenter liquid.
Agitation continually breaks the liquid
surface and increases surface area for
oxygen transfer.
 
 a. Sparged Stirred Tank
Bioreactors             

For liquid volumes greater than 3 L, air

sparging is required for effective oxygen
transfer.

Agitation: break up bubbles and increase

KLa.

Sparged fermenters required lower

agitation speeds for aeration efficiencies
comparable to non-sparged fermenters.
Air-sparged fermenters can have liquid
volumes of greater than 500,000 L
 4. BUBBLE DRIVEN BIOREACTORS

Sparging without mechanical agitation can

also be used for aeration and agitation.
     Two classes: 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.
 Airlift fermenters are more expensive
to construct than bubble column
reactors.

An airlift fermenter differs from bubble

column bioreactors by the presence of a
draft tube provides better mass and heat
transfer efficiencies and more uniform
shear conditions.
 a. Height to Diameter Ratios in Bubble
Driven Bioreactors

     Bubble driven fermenters are tall with

liquid height to base ratios of between 8:1
and 20:1.           
The tall design of these fermenters leads to
high gas hold-ups, long bubble residence
times and a region of high hydrostatic
pressure near the sparger at the base of the
fermenter.
2. Airlift Bioreactors - draft tube

    The main functions of draft tube:

1.  Increase mixing through the reactor

2.  The draft tube enhances axial
mixing throughout the whole reactor
3.   Reduce bubble coalescence.
Due to circulatory effect, draft tube induces in
the reactor. Circulation occurs in one
direction and hence the bubbles also travel in
one direction.
     Small bubbles lead to an increased
surface area for oxygen transfer.
Equalize shear forces throughout the
reactor (major reason why airlift
bioreactors have higher productivities
than stirred tank reactors).
5. AIRLIFT BIOREACTOR

a. Air-riser and Down-comer  

Three regions: air-riser, down-comer and
disengagement zone.
              Air-riser: region into which
bubbles are sparged (inside or outside of
draft-tube).
              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.
              Enhanced liquid circulation 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.

              Disengagement zone: add volume to

the reactor, reduce foaming minimize
recirculation of bubbles through the down
comer.
    Sudden widening at top of reactor slows
bubble velocity and disengages the bubbles
from the liquid flow. Carbon-dioxide rich
bubbles prevented from entering the down-
comer.

     Reduced bubble velocity in disengagement

zone leads to a reduction in loss of medium due
aerosol formation.

    Increase in area helps to stretch bubbles in

foams, causing the bubbles to burst.
    Axial flow circulation caused by the draft
tube also helps to reduce foaming.
Problems
 
      Higher energy requirements

    Higher levels of foam formation

excessive foaming and cell damage,
particularly with animal cell cultures. This
arises mainly due to the shear forces, which
arise bubble at the surface burst.
 
6. FIXED BED REACTORS
              Cells are immobilized by
absorption on or entrapment in solid,
non-moving solid surfaces(e.g: plastic
blocks, concrete blocks, wood shavings or
fibrous material such as plastic or glass
wool)..
 
              The liquid feed is either pumped
through or allowed to trickle over the
surface of the solids where the immobilized
cells convert the substrates into products.
             
Once steady state has been reached there
will be a continuous cell loss from the solid
surfaces.
             
Used in waste treatment and as
biological filters in small aquarium
water recycling systems.
              In other types of fixed-bed
fermenters, the cells are immobilized
in solidified gels such as agar or
carragenan.
 
              Cells are trapped inside pores of gels
(better cell retention and a large effective
surface area for cell entrapment).
             
Increase surface area for cell
immobilization: hollow fibres and pleated
membranes as immobilization surfaces.
             
Industrial applications: waste water
treatment, production of enzymes and
amino acids, steroid transformations
              Advantages: non-growing
cells can be used. Cells enzymatically
act on substrates in the feed. The
cells can be either inactivated or not
fed nutrients required for growth.
7.       PACKED BED BIOREACTOR
 
              Rate of mass transfer between cells
and medium depends on flow rate and on
thickness of biomass film on or near the
surface of the solid particles.
 
              Problems: poor mass transfer rates
and clogging.
 
              Used commercially with
enzymatically catalysts and with slowly or
non-growing cells.
              Used in anaerobic treatment of
high strength wastewaters (eg. food
processing wastes).
              Large plastic blocks are used as
solid supports for cells. These blocks have a
large surface area for cell immobilization and
when packed in the reactor, are difficult to
clog.
8. TRICKLE FLOW BIOREACTORS
              Class of packed bed reactors, in
which medium flows (or trickles) over the
solid particles. Particles are not immersed in
the liquid.
 
              Used in aerobic treatment of
sewage. Oxygen transfer is enhanced by
ensuring cells are covered by only a very thin
layer of liquid, thus reducing the distance
over which the dissolved oxygen must diffuse
to reach the cells.
The liquid medium
trickles over the surface
of the solids on which
the cells are immobilized

Oxygen diffuses through

a thin layer of liquid
surrounding the cells.
 Because stirring is not used,
considerable capital and energy
costs are saved. However, oxygen
transfer rates per unit volume are
low compared with sparged stirred
tank systems.
 
9. FLUIDIZED BED BIOREACTORS
 
 Maintaining high biomass
concentrations, good mass transfer rates
in continuous cultures.
  Mixing is assisted by the action of a
pump.
             

cells or enzymes are immobilized in and/or

on the surface of light particles.
              A pump located at base of tank
causes the immobilized catalysts to move
with the fluid. The pump pushes fluid and
particles in a vertical direction.
              Upward force of the pump is
balanced by the downward movement of
the particles due to gravity.
              This results in good circulation. 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 fluidized 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 microcarrier cultures
can be operated both in batch and
continuous mode.

Use of small particles increases the

surface area for cell immobilization and
mass transfer. Because the particles
are small and light, they can be easily
made to flow with the liquid (ie.
fluidized).
 2 Category: 2 phase system (not
aerated) and 3 phase system (aerated
by sparging).

              Fluidization of particles leads to the

surface of particles being continuously
turned over (increases mass transfer rate).
             
              Used in wastewater treatment.
Aquarium scale fluidized bioreactors for
biological nitrification are readily available.
 Used for animal cell culture. Animal
cells are trapped in gels or on the
surface of special particles known as
"microcarriers". Ex: perfusion culture
technology used for animal cell
 
culture.

6.10 INDUSTRIAL BIOREACTORS

              World’s largest fermenter (from
Chemical and Engineering News), 200 ft high
and 25 ft diameter.