DESIGN OF A FERMENTER

Fermentation Technology
Traditional Fermentation
Traditional Fermentation
Traditional Fermentation
Introduction

• A bioreactor is a vessel in which a substrate of

low value is utilized by living cells or enzymes to
generate a product of higher value.
• Bioreactors are extensively used for food
processing, fermentation, waste treatment, etc.
• Fermenters are the bioreactors which are
specially designed for fermentation.
• The main function of a fermenter is to provide a
controlled environment for the growth of
organisms, to obtain a desired product (e.g.
biomass, metabolite, bioconversion product)
The sizes of bioreactor can vary over several
orders of magnitudes
1. The microbial cell (few mm³)
2. Shake flask (100-1000 ml)
3. Laboratory fermenter (1-50 L)
4. Pilot scale (0.3-10 m³)
5. Plant scale (2-500 m³)
Considerations in Designing a Fermenter

1. The vessel should be capable of being operated

aseptically for a number of days and should be reliable
in long-term operation and meet the requirements of
containment regulations.
2. Adequate aeration and agitation should be provided to
meet the metabolic requirements of the microorganism.
However, mixing should not cause damage to the
organism nor cause excessive foam generation.
3. Power consumption should be as low as possible.
4. A system of temperature control, both during
sterilization and fermentation, should be provided.
5. A system of pH monitoring and control should be
provided together with the monitoring and control of
other parameters (e.g, dissolved oxygen, redox, etc.) as
appropriate.
6. Sampling facilities should be provided
Considerations in Designing a Fermentor

7. Evaporation losses from the fermenter should be

minimum.
8. The vessel should be designed to require the
minimal use of labor in operation, harvesting,
cleaning, and maintenance.
9. Ideally the vessel should be suitable for a range of
processes, but this may be restricted because of
containment regulations.
10.The vessel should be constructed to ensure smooth
internal surfaces, using welds
joints whenever possible.
Considerations in Designing a Fermentor

11.The vessel should be of similar geometry to both

smaller and larger vessels in the pilot plant or plant
to facilitate scale-up.
12.The cheapest materials, which enable satisfactory
results to be achieved should be used.
13.There should be adequate service provisions for
individual plants
Mechanically agitated and aerated fermenters
Mechanically agitated and aerated fermenters
Mechanically agitated and aerated fermenters
Fermenter Body Construction

• At pilot and industrial scale, when all fermenters

are sterilized in situ, any materials used will have
to be assessed on :
• their ability to withstand pressure
sterilization
• their ability to withstand corrosion
• their potential toxicity
• cost
Fermenter Body Construction

Glass fermenter
• In fermentations with strict aseptic requirements,
it is important to select materials that can
withstand repeated steam sterilization cycles.
• On a small scale (1 30 dm3) it is possible to use
glass and/or stainless steel.
• Glass is useful because it gives :
• smooth surfaces,
• nontoxic,
• corrosion proof,
• easy to examine the interior of the vessel.
Fermenter Body Construction

Glass fermenter
Fermenter Body Construction

Glass fermenter
Fermenter Body Construction

Stainless steel fermenter

• AISI (The American Iron and Steel Institute) grade 316
steels, which contain 18% chromium, 10% nickel and
2 2.5% molybdenum are now commonly used in
fermenter construction
• Chromium  enhances resistance to corrosion
(10 13% chromium develop an effective thin hydrous
oxide film on the metal)
• Nickel in high percent chromium steels  enhances
their corrosion resistance and improves their
engineering properties.
• Molybdenum  improves the resistance of stainless
steels to solutions of halogen salts and pitting by
chloride ions in brine or sea water
Fermenter Body Construction

Stainless steel fermenter

• The thickness of the

construction material
will increase with
scale.
• At 300,000 400,000
dm3 capacity, 7-mm
plate may be used for
the side of a vessel
and 10-mm plate for
the top and bottom,
which should be
hemispherical to
withstand pressures. Stainless Steel Pilot Plant Fermenters
(LSL, Luton, UK)
Fermenter Body Construction

Stainless steel fermenter Seals

• At this stage, it is important to consider the ways in which a
reliable aseptic seal is made between glass and glass,
glass and metal, or metal and metal joints.
• E.g. between a fermenter vessel and a detachable top or
base plate.
• This seal ensures that a good liquid- and/or gas-tight
joint is maintained in spite of the glass or metal expanding
or contracting at different rates with changes in
temperature during a sterilization cycle or an incubation
cycle.
• Nitryl or butyl rubbers are normally used for these seals as
they will withstand fermentation process conditions.
• These rubber seals have a finite life and should be checked
regularly for damage or perishing
Fermenter Body Construction

Stainless steel fermenter Seals

Achievement and Maintenance of
Aseptic Condition

• It will be necessary to be able to sterilize,

and keep sterile, a fermenter and its
contents throughout a complete growth
cycle.
• There may be also a need to protect
workers and the environment from
exposure to hazardous microorganisms or
animal cells.
• The containment requirements depend on
the size of the fermentation vessel
Achievement and Maintenance of
Aseptic Condition

• The following operations may have to be

performed to achieve and maintain aseptic
conditions and containment during a
fermentation:
1. Sterilization of the fermenter.
2. Sterilization of the air supply and the
exhaust gas.
3. Aeration and agitation.
4. The addition of inoculum, nutrients, and
other supplements.
5. Sampling.
6. Foam control.
7. Monitoring and control of various
parameters
Sterilization of The Fermenter

• Should be designed that it may be steam sterilized under

pressure.
• The medium may be sterilized in the vessel or separately,
and subsequently added aseptically.
• For in situ sterilization, the medium should be heated first
by coils or jacket to prevent formation of large amount of
condensate.
• Steam should be introduced through all the entry and exit
points except the air outlet from which steam should be
allowed to leave, as every point is a potential source of
contamination.
• All pipes should be constructed as simply as possible and
slope toward drainage points to make sure that steam
reaches all parts of the equipment
• For long-term aseptic operation welded joints should be
used wherever possible
Sterillization of The Air Supply

• Sterile air will be required in very large

volumes in many aerobic fermentation
processes.
• Two ways of sterilizing air :
1. Heat
2. Filtration
• Heat is generally too costly for full-
scale operation.
• Currently most fermenters are fitted
with cartridge-type filters. Made from
Polytetrafluoroethylene (PTFE), which
hydrophobic
Sterillization of The Air Supply

Packed Air Filter and Fermenter

Sterillization of The Air Supply

Packed Air Filter and Fermenter

• During sterilization the main nonsterile
air-inlet valve A is shut, and initially the
sterile air valve B is closed.
• Steam is applied at valve C and air is
purged downward through the filter to a
bleed valve at the base.
• When the steam is issuing freely through
the bleed valve, the valve B is opened to
allow steam to pass into the fermenter as
well as the filter.
Sterillization of The Air Supply

Simple Steam-Jacketed Air-Filter

Sterillization of The Air Supply

Simple Steam-Jacketed Air-Filter

• At the beginning of a sterilization cycle the valve A will be
closed and steam passed through valves B and C, and bled
out of D.
• Simultaneously steam will be passed into the steam jacket
through valve F and out of G.
• When steam is issuing freely from valve D, valve E may be
opened and steam circulated into the fermenter.
• The bleed valve D will have to be adjusted to ensure that
the correct pressure is maintained.
• Once the sterilization cycle is complete, valves B and E are
closed and A is opened to allow air to pass through the
heated filter and out of valve D to dry the filter.
• Finally the steam supply to the steam jacket is stopped.
• Valve D is closed and valve E opened, thus introducing
sterile air into the fermenter to achieve a slight positive
pressure in the vessel.
Sterillization of The Exhaust Gas

• Sterilization of the exhaust gas can be achieved by

0.2-µm filters on the outlet pipe.
• Under normal operation aerosol formation may occur
in the fermenter, and moisture and solid matter may
then plug the filter.
• To ensure satisfactory operation a cyclone separator
(for solids) and a coalescer (for liquids) would be
included upstream of two filters in series.
• The filters should be checked regularly to ensure that
no viable cells are escaping.
Sampling

• A sterile barrier must be maintained between the

fermenter contents and the exterior when the sample
port is not being used and it must be sterilizable after
use.

Simple Design for a Sampling Port An Alternative Simple Sample Port

Sampling

Simple Design for a Sampling Port

• A sample is obtained by removing the container of
formalin and closing valve A.
• Valves B and C are then opened until the piping has
been sterilized by steam.
• Valves B and C are then partially closed to allow a slow
stream of steam and condensate out of the sampling
port.
• Valve A is then opened slightly to cool the piping. The
broth is discarded.
• Valve C is then closed and a sample is collected.
• Valve A is then closed and the piping is resterilized
and left in the out of use arrangement.
Sampling

An Alternative Simple Sample Port

• Valves A and B being closed.
• Valve D is connected to a steam trap to avoid
condensate accumulation.
• To sterilize valve B prior to sampling, valve C is
partially closed, valve D completely closed and valve B
partially opened to allow a slow stream of steam and
condensate out of the sampling port.
•
broth is discarded.
• Valve C is then closed and a sample is collected. Valve
A is then closed and the piping is resterilized.
• In between collecting samples valves C and D are left
partially open
Feed Ports

• Additions of nutrients and acid/alkali to small fermenters

are normally made via silicone tubes which are autoclaved
separately and pumped after aseptic connection.
• In large fermenter units, the nutrient reservoirs and
associated piping are usually an integrated part which can
be sterilized with the vessel.
• However, there may also be ports which are used
intermittently.
• These can be sterilized in situ with steam after connection
has been completed and before any additions are made.
Sensor Probes

• have been used for many years

to provide an aseptic seal for glass electrodes in
stainless steel housings in fermenters.
• This system is suitable for lower level hazards,
provided that release of microorganisms is minimized.
• At higher containment levels, probes are fitted with
-ring seals.
• The use of pre-inserted backup probes is
recommended as a means for dealing with probe
failure rather than using a retractable electrode
housing during a fermentation cycle because of the
danger of leakage of broth
Foam Control

• When foaming becomes excessive, there is a

danger that filters become wet resulting in
contamination.
• Antifoams can be used to reduce foaming, but it
may cause problems with aeration or downstream
processing
• Foam breakers, which break down foam by an
impact mechanism created by some type of
rotating mechanism inside the fermenter.
Antifoaming agents
• Antifoaming agent / defoamer  reduce /
hinder the formation of foam on the
surface of liquids by reducing the surface
tension
• Examples : silicone oil, mineral oil,
vegetable oil, and polyoxyalkylene
derivatives
Industrial-scale penicillin
production by Penicillium
chrysogenum
Tower Fermenter

• Many types  main common feature : aspect ratio

(height : diameter)
• An elongated non-mechanically stirred fermenter
with an aspect ratio of at least 6 : 1 for the
tubular section
Bubble Column Fermenter

• Structurally very
simple and are not
mechanically
agitated.
• Aeration and
agitation are
achieved by gas
sparging and
consume less
energy than stirred
reactors.
Tower Fermenter

• Perforated plates were

positioned at intervals in the
tower to maintain maximum
yeast production.
• The settling zone  to provide a
zone free of rising gas so that
the cells could settle and return
to the main body of the tower.
• This design must be considered
as an intermediate between
single- and multistage systems.
• Towers of up to 20,000 dm3
capacity and capable of
producing up to 90,000 dm3
day 1 have been installed.

Tower Fermenter for the Brewing of Beer

Tower Fermenter
Cylindro-conical Vessels

• Application : brewing
• Cooling jacket around the
vessel upper part  to induce
convection mixing
• Cooling jacket around the
basal cone  to cause
flocculation and settling of
yeast after primary
fermentation
• Wort + yeast  40-48h
fermentation  mixing is
achieved by the generation of
CO2 that rises rapidly in the
vessel
Cylindro-conical Vessels

• Partially cleared beer left to allow secondary

fermentation & conditioning
• Advantages :
• Reduced process time (due to increase movement
within the vessel)
• Primary fermentation and conditioning (aging)
may be carried out in the same vessel
• The sedimented yeast may be easily removed
Air-lift Fermenters

• An air-lift fermenter is essentially a riser tube

(liquid ascending) where air is introduced
connected to a downcomer tube (liquid
descending)
Air-lift Fermenters

• Air or gas mixtures are introduced into the base

of the riser by a sparger during normal
operating conditions.
• The driving force for circulation of medium in
the vessel is produced by the difference in
density between the liquid column in the riser
(excess air bubbles in the medium) and the
liquid column in the downcomer (depleted in
air bubbles after release at the top of the loop).
Air-lift Fermenters

• Advantages :
• Simple design
• Easier sterilization (no curves of agitator)
• Low energy requirement (compared to stirred
tank)
• Greater heat removal
• Low cost
• Application : SCP (no need to remove heat
accumulated from stirring)
Deep-jet fermenters

• Injector : a jet of liquid is

surrounded by a jet of
compressed air (velocity :
5-100 m/s)
• Ejector : liquid jet enters
into larger gas-entrainer
• The air-medium mixture
falls down a slightly conical
shaft at a high velocity,
creates turbulence in the
fermenter
• Higher oxygen transfer rate
(compared to stirred vessel)