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SSG 2513

Introduction of Bioprocess
Engineering
5.0 Bioreactor selection and optimization
5.1 Identification of various bioreactor
5.2 Sterilization in bioreactor and bioprocess unit
operations

Goh Kian Mau

References:
Bioprocess Engineering. Shuler. (9.1, 9.2,
9.3, 9.5, 15.4.1,10.4)
Bioprocess Engineering Principles.
Pauline M. Doran. (P248.3 B56 1995)
Operation modes of bioreactors.
(TP.248.25.B55 O63 1992)
Bioreactor Design and product yield
(TP248.25 B55 B56 1992).

What is bioreactor?
The bioreactor in which biological/biochemical
reaction takes place occupies a central position
in the chemical process.
Synonymous for bioreactor: biochemical reactor,
biological reactor, fermenter, microbial reactor.
The reactions happens inside the vessel
because of the presence of either microbial
fermentation or enzyme (cell-free) reactions or
both.The cells or enzyme can be in suspension
or immobilized form.
Bioreactors can be operated as aerobic,
anaerobic, or solid state.

The bioreactor provides the volume (height, diameter) necessary for


the reaction and holds the amount of catalyst or cells required for the
reaction. The energy required to over-come the activation threshold
of each partial reaction is also supplied in the reactor, and the proper
parameters (temperature, concentration, pressure, pH, DO, etc) are
monitored and controlled.

Bioreactor Configurations
Batch Stirred Tank
only 70-80% of the volume of stirred
reactors is filled with liquid; this allows
adequate headspace for
disen-gagement (breaking off) of
droplets from the exhaust gas and to
accommodate any foam which may
develop. If foaming is a problem, a
supplementary impeller called a foam
breaker maybe installed as shown
above. Alternatively, antifoam agents
are added to the broth; because
reduces the rate of oxygen transfer.
Normally mechanical foam dispersal is
generally preferred.

Batch bioreactor
Exhaust air
(Air out)

Before inoculation (or adding the enzymes),


the batch reactor contains a certain volume
of nutrient (or substrate) in suspension.
After inoculation, the process is left
untouched, ie no material is added to or
removed from the reactor.
However, the reactor might be aerated.
Batch reactors are often referred to as
closed systems.
The term 'closed' refers to the fact that
material can neither enter nor leave the
reactor. (exceptional: airin and airout)

Air in

Batch bioreactor-ideal mixing


In an ideal well-mixed
bioreactor, the mixing is
assumed to be intense
enough that the fluid is
homogeneous through the
reactor.
no dead zones or clumps
of undissolved solid
substrate floating around
and there is no
concentration gradients in
vessel.

Fed batch bioreactor


The fed-batch reaction (FBR) is a batch
reactor to which, when the nutrients
approach depletion, fresh nutrients are
added. In other words, the reactor is fed. It
is assumed that the concentration of the
nutrients added is so high that volume
changes are negligible (justifying the batch
part of the name).

Bubble column
- no mechanical agitation.
-aeration and mixing are achieved by gas
sparging; this requires less energy than
mechanical stirring.
-height-to-diameter ratio is usually high to
maximize the aeration mixing
-Advantages of bubble columns include low
capital cost, lack of moving parts, and
satisfactory heat- and mass-transfer
perfor-mance. As in stirred vessels,
foaming can be a problem requiring
mechanical dispersal or addition of
antifoam to the medium. (Your imagination
is needed, bubble and foam is different!)

Bubble column
The oxygen transfer from the air bubble
and the mixing inside the bubble-column
depend entirely on the behaviour of the
bubbles released from the sparger.
Homogeneous flow occurs only at low
gas flow rates and when bubbles
leaving the sparger are evenly
distributed across the column crosssection. In homogeneous flow, all
bubbles rise with the same upward
velocity and there is no backmixing of the
gas phase.
Liquid mixing in this flow regime is also
limited, arising solely from entrainment in
the wakes of the bubbles. Under normal
operating conditions at higher gas
velocities, large chaotic circulatory flow
is develop and heterogeneous flow
occurs as illustrated in figure on the left.

Their distinguishing feature


compared with the bubble
column is that of liquid flow
are more defined owing to
the physical separation of
up-flowing and down-flowing
streams.
gas is sparged into a part of
the vessel section called the
riser.
Gas disengages at the top
of the vessel leaving heavier
bubble-free liquid to
recirculate through the
downcomer.
Liquid circulates in airlift
reactors as a result of the
density difference between
the riser and downcomer.

Airlift bioreactor

Packed-bed reactors are


used with immobilised or
particulate biocatalysts.
Medium/buffer can be fed
either at the top or bottom
of the column and forms a
continuous liquid phase
between the particles.
Damage due to particle
wear is minimal in packed
beds compared with stirred
reactors.

Packed-bed
bioreactor

Normally the vessel is not fully


packed with the bead for the
expansion and movement of the
bed.
When packed beds are
operated in upflow mode with
beads of appropriate size and
density, the bed expands at
liquid flow rates due to upward
motion of the particles.
Because particles in fluidized
beds are in constant motion,
channeling and clogging of the
bed are avoided and air can be
introduced directly into the
column.

Fluidized bed
bioreactor

Trickle-bed reactor is
another variation of
the packed bed.
Liquid is sprayed onto
the top of packing and
trickles down through
the bed in small
rivulets.
Air may be introduced
at the base. Tricklebed -bioreactors are
used widely for
aerobic wastewater
treatment.

Trinkle-bed
bioreactor

Membrane bioreactor (MBR)


In a membrane reactor continuous processing is
possible if production and separation can be
incorporated in one system.
One application of the membrane bioreactor is to
remove product that is formed, especially for those
that can inhibit the enzyme reaction.

Sterilization in bioreactor
Most bioreactors (especially fermentation process and
production of pharmaceutical products) outside of the
food and beverage industry are carried out using aseptic
conditions.
Most industrial bioreactors are designed for in situ steam
sterilisation under pressure. The vessel should have a
minimum number of internal structures, ports, nozzles,
connections and other attachments to ensure that steam
reaches all parts of the equipment. For effective
sterilisation, all air in the vessel and pipe connections
must be displaced by steam.
Think: In industry, where does the steam supply come
from?

Example of steam sterilization at the sampling port


of a fermentor
Sampling ports are fitted to fermenters to
allow removal of broth for analysis.
Initially, valves A and D are closed;
valves B and C are open to maintain a
steam barrier between the reactor and
the outside environ-ment.
Valve C is then closed, valve B partially
closed and valve D partially opened to
allow steam and condensate to bleed
from the sampling port D.
For sampling, A is opened briefly to cool
the pipe and carry away any condensate
that would dilute the sample; this broth is
discarded.
Valve B is then closed and a sample
collected through D. When sampling is
complete, valve A is closed and B
opened for re-sterilisation of the sample
line; this prevents any contaminants
which entered while D was open from
travelling up to the fermenter. Valve D is
then closed and valve C re-opened.

Sterilization in other unit operation in


bioprocess industry

Heat sterilization
Chemical sterilization (refer to handouts)
Sterilization using radiation
-Radiation sterilization is a good alternative for
sterilizing disposable medical, laboratory and
pharmaceutical products. The radiation that are
commonly are ultraviolet 200-280 nm (UV), -radiation
and -radiation.
In some industries such as the pharmaceutical, high
quality of water (such as RO, milli-Q, water-forinjection (WFI)) is required. Normally, a series of
filtration and ion-exchange purification steps are
involved. High quality water are normally keep in
water tanks before utilize, and these tanks usually are
equipped with UV radiation to avoid the growth of
microorganism.

Sterilization in other unit operation in


bioprocess industry

Sterilization by using membrane filter


All microorganism including viruses have certain sizes.
The microorganism can simply be removed by using
membrane filter that has pore size smaller than
microorganism. In some application, a series of
membrane filters are used.
Depth filters with bigger pore size (e.g 0.6 m) may be
used first to remove insoluble particle prior passing
through membrane filters with smaller pore size (e.g
0.45 m or 0.2 m). (Think: Why? Check inside
textbook what is the size of common microorganism).

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