Range of Fermentation Processes
To Produce Microbial cells or Biomass
To Produce Microbial Enzymes
To Produce Microbial Metabolites
To Produce Recombinant Products
To modify a compound which is added to the
fermentation (Transformation)
Basic Design of a Fermenter
Various components of an ideal fermenter for batch process are:
Monitoring and controlling parts of fermenter are:
Basic Functions of a Fermenter
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.
Adequate aeration and agitation should be provided to meet the
metabolic requirements of the micro-organism. However, the mixing
should not cause damage to the organism.
Power consumption should be as low as possible.
A system of temperature control should be provided.
A system of pH control should be provided.
Sampling facilities should be provided.
Evaporation losses from the fermenter should not be excessive.
The vessel should be designed to require the minimal use of
labour in operation, harvesting, cleaning and maintenance.
Ideally the vessel should be suitable for a range of processes, but
this may be restricted because of containment (the action of
keeping something harmful under control or within limits)
regulations.
The vessel should be constructed to ensure smooth
internal surfaces, using welds instead of flange joints
whenever possible.
The vessel should be of similar geometry to both
smaller and larger vessels in the pilot plant or plant to
facilitate scale-up.
The cheapest materials which enable satisfactory
results to be achieved should be used.
There should be adequate service provisions for
individual plants
Hazard Assessment Systems
Once the organism has been allocated to a hazard group, the
appropriate containment requirements can be applied.
Hazard group 1 organisms used on a large scale only require Good
Industrial Large Scale Practice (GILSP).
Processes in this category need to be operated aseptically but no
containment steps are necessary, including prevention of escape of
organisms.
If the organism is placed in Hazard group 4 the stringent requirements
of level 3 will have to be met before the process can be operated.
Materials for Body Construction of a 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 to 30 dm3) it is possible to use glass
and/or stainless steel.
Glass is useful because it gives smooth surfaces, is
non-toxic, corrosion proof and it is usually easy to
examine the interior of the vessel.
Two basic types of Fermenters
A glass vessel with a round or flat bottom and a top
flanged carrying plate.
All vessels of this type have to be sterilized by
autoclaving.
A glass cylinder with stainless-steel top and bottom
plates.
Vessels with two stainless steel plates cost
approximately 50% more than those with just a top
plate.
At pilot and large scale, when all fermenters are
sterilized in situ, any materials used will have to be
assessed on their ability to withstand pressure
sterilization and corrosion and on their potential
toxicity and cost.
Pilot scale and Industrial scale vessels are normally constructed of
stainless steel or at least have a stainless-steel cladding to limit
corrosion.
The American Iron and Steel Institute (AISI) states that steels
containing less than 4% chromium are classified as steel alloys and
those containing more than 4% are classified as stainless steels.
Mild steel coated with glass or phenolic epoxy materials has
occasionally been used
The corrosion resistance of stainless steel is thought to depend on
the existence of a thin hydrous oxide film on the surface of the
metal.
The film is stabilized by chromium and is considered to be
continuous, non-porous, insoluble and self healing.
If damaged, the film will repair itself when exposed to air or an
oxidizing agent.
Types of Seals
Gasket Seal
Lip Seal
O ring Seal
With glass and metal, a seal can be made with a
compressible gasket, a lip seal or an '0' ring.
With metal to metal joints only an '0' ring is suitable.
Nitryl or butyl rubbers are normally used for these
seals as they will withstand fermentation process
conditions.
A single '0' ring seal is adequate for GILSP and levels 1
and B2, a double '0' ring seal is required for levels 2
and B3 and a double '0' ring seal with steam between
the seals (steam tracing) is necessary for levels 3 and
B4
Aeration and Agitation
Primary purpose of aeration is to provide
microorganisms in submerged culture with sufficient
oxygen for metabolic requirements.
While agitation should ensure that a uniform
suspension of microbial cells is achieved in a
homogenous nutrient medium.
Agitator (Impellers)
The agitator is required to achieve a number of mixing objectives,
e.g. bulk fluid and gas-phase mixing, air dispersion, oxygen transfer,
heat transfer, suspension of solid particles and maintaining a
uniform environment throughout the vessel contents.
Agitators may be classified as disc turbines, vaned discs, open
turbines of variable pitch and propellers.
The disc turbine consists of a disc with a series of rectangular vanes
set in a vertical plane around the circumference.
The vaned disc has a series of rectangular vanes attached vertically
to the underside.
Air from the sparger hits the underside of the disc and is displaced
towards the vanes where the air bubbles are broken up into smaller
bubbles.
Stirrer Glands and Bearings
The satisfactory sealing of the stirrer shaft assembly
top plate has been one of the most difficult problems
to overcome in the construction of fermentation
equipment which can be operated aseptically for long
periods.
The stirrer shaft can enter the vessel from the top, side
or bottom of the vessel.
A simple stirrer seal
A simple Stirrer Seal
A porous bronze bearing for a 13-mm shaft was fitted
A simple Stirrer Seal
in the centre of the fermenter top and another in a
A simple Stirrer Seal
yoke directly above it.
The bearings were pressed into steel housings, which
screwed into position in the yoke and the fermenter
top.
The lower bearing and housing were covered with a
skirt-like shield having a 6.5 mm overhang which
rotated with the shaft and prevented air-borne
contaminants from settling on the bearing and
working their way through it into the fermenter.
Four Main Types
The Stuffing Box (Packed Gland seal)
The Mechanical Seal
Simple Bush Seal
The Magnetic Drive Seal
The Stuffing Box (Packed Gland Seal)
The shaft is sealed by several layers of packing rings
of asbestos or cotton yarn, pressed against the shaft by
a gland follower.
Chain et al. (1954) used two stuffing boxes on the
agitator shaft with a space in between kept filled with
steam.
These seals are sufficient for the requirements of
GILSP containment.
The Mechanical Seal
The seal is composed of two parts, one part is
stationary in the bearing housing, the other rotates on
the shaft, and the two components are pressed
together by springs or expanding bellows.
The two meeting surfaces have to be precision
machined, the moving surface normally consists of a
carbon-faced unit while the stationary unit is of
stellite-faced stainless steel.
Magnetic Drive
The problems of providing a satisfactory seal when the
impeller shaft passes through the top or bottom plate of
the fermenter may be solved by the use of a magnetic
drive in which the impeller shaft does not pierce the
vessel.
A magnetic drive consists of two magnets: one driving
and one driven.
The driving magnet is held in bearings in a housing on
the outside of the head plate and connected to a drive
shaft.
The internal driven magnet is placed on one end of the
impeller shaft and held in bearings in a suitable housing
on the inner surface of the headplate.
Baffles
Four baffles are normally incorporated into agitated vessels of
all sizes to prevent a vortex and to improve aeration efficiency.
Baffles are metal strips roughly one-tenth of the vessel diameter
and attached radially to the wall.
The agitation effect is only slightly increased with wider baffles,
but drops sharply with narrower baffles.
It is recommended that baffles should be installed so that a gap
existed between them and the vessel wall, so that there was a
scouring action around and behind the baffles thus minimizing
microbial growth on the baffles and the fermenter walls.
Extra cooling coils may be attached to baffles to improve the
cooling capacity of a fermenter without unduly affecting the
geometry.
Aeration System (Spargers)
A sparger may be defined as a device for introducing air into the liquid in a fermenter.
Three basic types of sparger have been used and may be described as the Porous
sparger, the Orifice sparger (a perforated pipe) and the Nozzle sparger (an open or
partially closed pipe).
Porous Sparger
The porous sparger of sintered glass, ceramics or metal, has been used primarily on a
laboratory scale in non-agitated vessels.
The bubble size produced from such spargers is always 10 to 100 times larger than the
pore size of the aerator block.
There is also the problem of the fine holes becoming blocked by growth of the
microbial culture.
Orifice Sparger
In small stirred fermenters the perforated pipes were arranged below the impeller
in the form of crosses or rings (ring sparger), approximately three-quarters of the
impeller diameter.
In most designs the air holes were drilled on the under surfaces of the tubes
making up the ring or cross.
ger holes should be at least 6 mm (1/4 inch) diameter because of the tendency of
smaller holes to block and to minimize the pressure drop.
Nozzle Sparger
Single open or partially closed pipe as a sparger to provide the stream of air bubbles
Ideally the pipe should be positioned centrally below the impeller and as far away
as possible from it to ensure that the impeller is not flooded by the air stream.
Sterilization of Air Supply for Fermentation
Sterile air will be required in very large volumes in
many aerobic fermentation processes.
Heating and Filtration are the main methods for
sterilization. Heat is generally too costly for full scale
operation.
Glass wool, glass fibre or mineral slag wool have been
used as filter material, but currently most fermenters
are fitted with cartridge-type filters.
Two Procedures based on the construction of filter unit.
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 downwards
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.
It is essential to adjust the bleed valve to ensure that the
correct sterilization pressure is maintained in the
fermenter and filter for the remainder of the sterilization
cycle.
Valves and Steam Traps
Valves attached to fermenters and ancillary equipment are used
for controlling the flow of liquids and gases in a variety of ways.
There are four main types of valves:
Simple ON/OFF valves which are either fully open or fully closed.
Valves which provide coarse control of flow rates.
Valves which may be adjusted very precisely so that flow rates
may be accurately controlled.
Safety valves which are constructed in such a way that liquids or
gases will flow in only one direction.
Gate Valves
In this valve, a sliding disc is moved in or out of the flow path by turning
the stem of the valve.
It is suitable for general purposes on a steam or a waterline for use when
fully open or fully closed and therefore should not be used for regulating
flow.
Not suitable for aseptic conditions
there may be leakage round the stem of the valve which is sealed by a
simple stuffing box.
This means that the nut around the stem and the packing must be checked
regularly.
Globe Valves
In this valve, a horizontal disc or plug is raised or
lowered in its seating to control the rate of flow.
It is not suitable for aseptic operation because of
potential leakage round the valve stem which is similar
in design to that of the gate valve.
There is a high pressure drop across the valve because
of the flow path.
Piston Valves
The piston valve is similar to a globe valve except that
flow is controlled by a piston passing between two
packing rings.
This design has proved in practice to be very efficient
under aseptic operation.
There may be blockage problems with mycelial culture
and the pressure drop is similar to a globe valve.
Needle Valve
The needle valve is similar to the globe valve, except
that the disc is replaced by a tapered plug or needle
fitting into a tapered valve seat.
The valve body can be used to give fine control of
steam or liquid flow.
Accurate control of flow is possible because of the
variable orifice formed between the tapered plug and
the tapered seat.
The aseptic applications are very limited.
Ball Valve
This valve has been developed from the plug valve.
The valve element is a stainless-steel ball through
which an orifice is machined.
The ball is sealed between two wiping surfaces which
wipe the surface and prevent deposition of matter at
this point.
The valve is suitable for aseptic operation, can handle
mycelial broths and can be operated under high
temperatures and pressures.
Butterfly Valve
The butterfly valve consists of a disc which rotates
about a shaft in a housing.
The disc closes against a seal to stop the flow of liquid.
This type of valve is normally used in large diameter
pipes operating under low pressure where absolute
closure is not essential. It is not suitable for aseptic
operation.
Diaphragm Valve
Like the pinch valve, the diaphragm valve makes use of
a flexible closure, with or without a weir.
Suitable for aseptic operation.
Most Suitable Valve
Among these group of valves which have just been
described, globe and butterfly valves are most commonly
used for ON/OFF applications, gate valves for crude flow
control, needle valves for accurate flow control and ball,
pinch or diaphragm valves for all sterile uses.
Check Valves
The purpose of the check valve is to prevent
accidental reversal of flow of liquid or gas in a pipe due
to breakdown in some part of the equipment.
There are three basic types of valve: swing check, lift
check and combined stop and check with a number of
variants.
The swing check valve is most commonly used in
fermenter designs.
Steam Traps
In all steam lines it is essential to remove any steam
condensate which accumulates in the piping to ensure
optimum process conditions
This may be achieved by incorporating steam traps,
which will collect and remove automatically any
condensate at appropriate points in steam lines
