BIOREACTOR

ENGINEERING

BIOREACTOR
A bioreactor is a reactor system used
for the culture of LIVING CELLS. They
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. They
similarly vary in cost from a few cents
to a few million dollars.
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BIOREACTORS
Standing Cultures
Shake flasks
Mechanically stirred bioreactors
Bubble driven bioreactors

Airlift bioreactors
Packed bed and trickle flow
bioreactors
Fluidized bed bioreactors

Standing Cultures
Standing cultures - T flasks
T-flasks used in the small scale
culture of animal cells are another
example of a standing culture. Tflasks are normally incubated
horizontally to increase the surface
area for oxygen transfer.
The surface aeration rate in
standing cultures can be increased
by using large volume flasks.
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Standing Cultures
Fernback flasks----Have larger surface area
The following photograph shows a 3 litre
"Fernback" flask containing 1 litre of medium
and a 250 ml Erlenmeyer flask containing
100 ml of medium. Note how the former has
a large surface area.
Large Pyrex flasks are used for the small
scale production of fermented products. One
example is Kombucha tea which is a tea
brewed by mixture of yeasts and acetic acid
bacteria.
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Shake flasks
Shake flasks are commonly used for
small scale 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.9

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Mechanically Stirred Bioreactors

For liquid volumes greater than 3 liters,
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 nonsparged fermenters. Air-sparged
fermenters can have liquid volumes of
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greater than 500,000 liters.

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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.
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Airlift fermenters are however

more expensive to construct than
bubble column reactors.
An airlift fermenter differs from
bubble column bioreactors by the
presence of a draft tube which
provides
>better mass and heat transfer
efficiencies
>more uniform shear conditions.
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An airlift fermenter differs from a bubble

column bioreactor by the presence of a draft
tube. The main functions of the draft tube
are to:
Increase mixing through the reactor 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. 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.
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 Equalize shear forces throughout the

reactor. This is believed to be the major
reason why airlift bioreactors have higher
productivities than stirred tank reactors.

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Airlift bioreactor
An air-lift reactor is divided into three regions:
the air-riser, down-comer and disengagement
zone.
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.

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Airlift bioreactor
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 downcomer. 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.
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Immobilized Cell
Reactors

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Fixed bed reactors

In fixed bed fermenters, the cells are
immobilized by absorption on or
entrapment in solid, non-moving solid
surfaces.

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In one type of fixed bed fermenter, the

cells are immobilized on the surfaces
of immobile solid particles such as
plastic blocks
concrete blocks
wood shavings or
fibrous material such as plastic or
glass wool.
*In other types of fixed-bed fermenters,
the cells are immobilized in solidified
gels such as agar or carrageenin.
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**Industrial applications of
fixed bed reactors include
waste water treatment
production of enzymes and
amino acids
steroid transformations

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Cell recycle systems

In a fermenter with cell recycle the cells
are separated from the effluent and then
recycled back to the fermenter; thus
minimizing cell removal from the fermenter
Cell recycle is used in activated sludge
systems. A portion of the cells are
separated in a settling tank and returned
to the activated sludge fermenter.

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Packed and Trickle bed Bioreactors

Packed bed reactors often suffer from
problems caused by poor mass transfer
rates and clogging. Despite this they are
used commercially with enzymatic catalysts
and with slowly or non-growing cells.
Trickle bed reactors are a class of packed
bed reactors in which the medium flows (or
trickles) over the solid particles. In these
reactors, the particles are not immersed in
the liquid. They are used widely in aerobic
treatment of sewage.
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Fluidized bed reactors

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.

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Aeration and Agitation in

Animal Cell Bioreactors
Animal cells fragilecan be easily
damaged
In mammalian cell culture the objectives of
maximizing oxygen transfer must be
balanced by the need to maintain cell
integrity. Fortunately, mammalian cell
growth rates are considerably slower than
those of most aerobic microorganisms and
oxygen transfer requirements are
therefore also proportionately lower.
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Methods of minimizing cell damage

Bubble free oxygenation - External
oxygenation
A more commonly used and effective
method of bubble free oxygenation is to
use a separate oxygenation chamber.
The medium is oxygenated in a separate unit
which can either be a stirred tank reactor or a
static mixer. The oxygenated medium is
pumped into the bioreactor while the oxygen
depleted medium is pumped back into the
oxygenation unit
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 The same principle can be used

with immobilized cell cultures
such as fluidized bed and fibre
bed reactors.

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Effect of eddy streamlines on cell

> 2/3-1/2 bead size
Power near impeller
= mass of fluid near impe
= Di3

Kolmogrov Scale

= (3/)1/4

Rate of energy dissipation or

energy dissipated per unit mass of fluid

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Kinematic viscosity

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