1

Bioreactors (Fermenters): Function, Designs and types

A bioreactor is a device in which a substrate of low value is utilized by living cells or

enzymes to generate a product of higher value. Bioreactors arc extensively used for food
processing, fermentation, waste treatment, etc.

On the basis of the agent used, bioreactors are grouped into the following two broad
classes: (i) those based on living cells and, (ii) those employing enzymes. But in terms of
process requirements, they are of the following types: (i) aerobic, (ii) anaerobic, (iii) solid
state, and (iv) immobilized cell bioreactors.

All bioreactors deal with heterogeneous systems dealing with two or more phases, e.g.,
liquid, gas, solid. Therefore, optimal conditions for fermentation necessitate efficient
transfer of mass, heat and momentum from one phase to the other. Chemical engineering
principles are employed for design and operation of bioreactors. But, in general,
theoretical explanation usually lags behind technical realization.

A bioreactor should provide for the following: (i) agitation (for mixing of cells and
medium), (ii) aeration (aerobic fermenters; for O2 supply), (iii) regulation of factors like
temperature, pH, pressure, aeration, nutrient feeding, liquid level, etc., (iv) sterilization
and maintenance of sterility, and (v) withdrawal of cells/medium (for continuous
fermenters). Modern fermenters are usually integrated with computers for efficient
process monitoring, data acquisition, etc.

The first truly large-scale aseptic anaerobic fermentation vessels were developed in the
wake of the process developed (during the First World War, 1914-1918) by Weizmann and
co-workers of U.K. to produce acetone by a deep liquid fermentation using Clostridium
acetobutylicum.

For this, large cylindrical vessels of mild steel that permitted sterilization with steam
under pressure were constructed, and piping, joints and valves were specially designed to
maintain aseptic conditions, which were the major problem; mixing was achieved by the
large volumes of gas produced during fermentation.
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The large-scale aerobic fermentation vessels were first used in Central Europe during
1930s for the production of compressed yeast; these fermenters had large cylindrical
tanks in which air was introduced at the base via a network of perforated pipes.

In later modifications, mechanical impellers were used to improve mixing of broth and
dispersal of air bubbles. Fermenter design was considerably improved during 1940s to
accommodate the requirements of strict aseptic conditions, and good agitation and
aeration for penicillin production from submerged cultures; for this, steel fermenters with
working volumes of 54,000 1 were built in U.S.A. In 1944, Cooper and co-workers (and
several others) reported the findings from studies on agitation in baffled stirred tank
fermenters, which had a major influence on the design of later fermenters.

Basic Functions of a Fermenter:

1. It should provide a controlled environment for optimum biomass/product yields.

2. It should permit aseptic fermentation for a number of days reliably and dependably,
and meet the requirements of containment regulations. Containment involves prevention
of escape of viable cells from a fermenter or downstream processing equipment into the
environment.

3. It should provide adequate mixing and aeration for optimum growth and production,
without damaging the microorganisms/cells. The above two points (items 2 and 3) are
perhaps the most important of all.

4. The power consumption should be minimum.

5. It should provide easy and dependable temperature control.

6. Facility for sampling should be provided.

7. It should have a system for monitoring and regulating pH of the fermentation broth.

8. Evaporation losses should be as low as possible.

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9. It should require a minimum of labour in maintenance, cleaning, operating and

harvesting operations.

10. It should be suitable for a range of fermentation processes. But this range may often
be restricted by the containment regulations.

11. It should have smooth internal surfaces, and joints should be welded wherever
possible.

12. The pilot scale and production stage fermenters should have similar geometry to
facilitate scale-up.

13. It should be contrasted using the cheapest materials that afford satisfactory results.

14. There should be adequate service provisions for individual plants.

Fermenter Design:

A bioreactor is a device 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.

On the basis of the agent used, bioreactors are grouped into the following two broad
classes: (i) those based on living cells and, (ii) those employing enzymes. But in terms of
process requirements, they are of the following types: (i) aerobic, (ii) anaerobic, (iii) solid
state, and (iv) immobilized cell bioreactors.

All bioreactors deal with heterogeneous systems dealing with two or more phases, e.g.,
liquid, gas, solid. Therefore, optimal conditions for fermentation necessitate efficient
transfer of mass, heat and momentum from one phase to the other. Chemical engineering
principles are employed for design and operation of bioreactors.

But, in general, theoretical explanation usually lags behind technical realization. A

bioreactor should provide for the following: (i) agitation (for mixing of cells and medium),
(ii) aeration (aerobic fermenters; for O2 supply), (iii) regulation of factors like temperature,
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pH, pressure, aeration, nutrient feeding, liquid level, etc., (iv) sterilization and
maintenance of sterility, and (v) withdrawal of cells/medium (for continuous fermenters).
Modern fermenters are usually integrated with computers for efficient process monitoring,
data acquisition, etc.

Agitation and Aeration:

In scaling up, both chemical (O2, pH, medium constituents and removal of wastes) and
physical (the configuration of bioreactor and power supplied to the reactor) factors have
to be optimised for good results.

The medium must be suitably stirred to keep the cells in suspension and to make the
culture homogeneous; it becomes increasingly difficult with the scaling up. Various types
of stirrers range from simple magnetic stirrers, flat blade turbine impellers, to marine
impellers, to those using pneumatic energy, e.g., airlift fermenter, and those using
hydraulic energy, e.g., medium perfusion.

Improved mixing can be obtained by changing the design of stirrer paddle or by using
multiple impellers. The objective of stirring is to achieve good mixing without causing
damage to the cells. Vibro-mixer achieves stirring by vertical reciprocating motion of 0.1-
3 mm at a frequency of 50 cycles/sec of a mixing disc fixed horizontally to the agitator
shaft. These stirrers cause random mixing, less foaming and lower shear forces.

It is important to supply sufficient O2 without damaging the cells. Mean O2 utilization rate
by cells is about 6 mg O2/106 cells/hour. But O2 is only sparingly soluble in culture
medium; the oxygen transfer rate (OTR) from gas phase into medium is about 17
g/cm/hr.

Therefore surface aeration can support about 50 x 106 cells in I 1 culture vessel. Efficient
aeration is achieved by bubbling air through the medium (sparging), but this may
damage animal cells due to the high surface energy of the bubble and on the cell
membrane.
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The damage can be reduced by using larger bubbles, lower gassing rates and by adding
non-nutritional supplements like Pluronic F-68 (polyglycol) and sodium carboxymcthyl
cellulose (these protect cells from damage due to shear forces and bubbles, respectively).
Silicone tubing (highly gas permeable) can be arranged inside the culture vessel (2-5 cm
tubing of 30 111 length for a 1000 1 culture) and air is passed though the tube; however
it is inconvenient to use.

Aeration may be achieved by medium perfusion, in which medium is continuously taken

from culture vessel, passed through an oxygenation chamber and returned to the culture.
The cells are removed from the medium taken for perfusion so that the medium can be
suitably altered, e.g., for pH control. Perfusion is used with glass bead and, more
particularly, with micro-carrier systems.

Where considered safe and desirable, O2 supply in the culture vessel can be enhanced
from the normal 21% to a higher value and the air pressure can be increased by 1
atmosphere. This increases the O2 solubility and diffusion rates in the medium, but there
is a risk of O2 toxicity.

The basic objective of aeration is to provide microorganisms growing in submerged

cultures with adequate oxygen for their metabolic needs. Agitation, on the other hand,
aims to ensure a homogeneous distribution of microorganisms and the nutrients in the
broth.

The type of aeration- agitation system used in the fermenter is dictated by the
characteristics of the fermentation process. For example, in processes based on low
viscosity, low total solids broths, agitation may not be needed as aeration itself would
create the necessary agitation.

Fine bubble aerators without mechanical agitation offer the advantage of lower
equipment and power costs. Such fermentations are usually carried out in vessels having
height/diameter ratio of 5 : 1, but a tall column of liquid would require a higher energy
input for the compression of air used for aeration.
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However, mechanical agitation is usually necessary for fermentation processes based on

actinomycetes and fungi. The following components of the fermenter are required for
aeration and agitation: (t) agitator (impeller), (ii) stirrer glands and bearings, (iii) baffles,
and (iv) sparger (the aeration system).

1. Agitator (Impeller):

Agitators achieve the following objectives; (a) bulk fluid and gas-phase mixing, (b) air
dispersion, (c) oxygen transfer, (d) heat transfer, (e) suspension of solid particles, and (f)
maintenance of a uniform environment throughout the vessel.

These objectives are achieved by a suitable combination of the most appropriate agitator,
air sparger and baffles, and the best positions for nutrient feeds, acid or alkali for pH
control and antifoam addition. Agitators are of several different types, e.g., (i) disc
turbines, (ii) vaned discs, (iii) open turbines of variable pitch and (iv) propellers.

Disc turbine consists of a disc with a series of rectangular vanes set in a vertical plane
around its perpheri (Fig. 14.1 A). The vaned disc turbine has a series of rectangular vanes
attached vertically to the underside of the disc (Fig. 14. 1B). In case of variable pitch open
turbine, the vanes are attached directly to a boss on the agitator shaft (Fig. 14.1C).

The marine propeller is similar to variable pitch open turbine, except that it has blades in
the place of vanes (Fig. 14.1D). In case of disc and vaned disc turbines, the air bubbles
from the sparger first-hit the underside of disc before being broken into smaller bubbles
and dispersed by the vanes.

But in the case of the latter two types of agitators, air bubbles contact the vanes/blades
directly and arc broken up and dispersed by them. These basic agitation devices have
been variously modified. For example, the variable pitch open turbine scheme has been
modified to develop four modern agitator types, viz., Scaba 6SRGT, Prochem Maxflo T,
Lightning A315 and the Ekato Intcrmig.

The Rushton disc turbine, having a diameter of one-third the fermenter diameter, has
been long considered optimum for many fermentation processes. The disc turbine was
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considered optimum because it was shown to be able to break up a fast air stream
without itself becoming flooded in air bubbles; the latter situation seriously hampers
oxygen dispersal in the broth.

In contrast, the impeller and open turbine were found to have the tendency to be flooded
in air at higher aeration rates. In subsequent studies, it was found that in low viscosity
broths, all the four agitator types can achieve good gas dispersion provided the agitator
speed is high enough.

In such broths, agitator type does not appear to be a significant factor affecting oxygen
transfer efficiencies. In high viscosity broths, however, gas dispersal presents problems
and is greatly reduced. In view of this, a number of agitators have been developed for
high viscosity broths, e.g., Scaba 6SRGT, Prochem Maxflow T, Lightning A315 and Ekato
Intermig (Fig. 14.2).

These agitators are larger, require lower power input (they do not lose as much power as
the Rushton turbines when aerated), are able to handle higher air volumes without
flooding, and give better bulk blending and heat transfer in more viscous media.

But they can cause mechanical problems mostly of vibrational nature. Good mixing and
aeration in high viscosity broths may also be achieved by a dual impeller combination in
which the lower impeller primarily dispenses the air, while the upper impeller primarily
enhances mixing of the broth.

2. Stirrer Glands and Bearings:

The satisfactory sealing of the stirrer shaft assembly has been one of the most difficult
problems; this is very important for maintaining aseptic conditions over long periods. Four
basic types of seal assembly have been used in fermenters: (1) the stuffing box (packed-
gland seal), (2) the simple bush seal, (3) the mechanical seal and (4) the magnetic drive.
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Most modern fermenters use mechanical seals; these

seals are more expensive, but they are more durable
and less prone to leakage or contaminant entry.
Magnetic drives, although quite expensive, are being
used in some animal cell culture vessels.

The mechanical
seal consists of
two parts; one
part remains
stationery in the
bearing housing,
while the other
rotates on the
shaft. The two
components of the seal are pressed together
by springs or expanding bellows. Steam
condensate is used to lubricate and cool the
seals during operation and servers as a
contaminant barrier.

3. Baffles:

Baffles are metal strips roughly one-tenth of the

vessel diameter and attached radially to the fermenter wall (Fig. 14.3). They are normally
used in fermenters having agitators to prevent vortex formation and to improve aeration
efficiency.

Usually, four baffles are used, but larger fermenters may have 6 or 8 baffles. Extra
cooling coils may be attached to baffles to improve cooling. Further, the baffles may be
installed in such a way that a gap exists between the baffles and the fermenter wall. This
would lead to a scouring action around and behind the baffles, which would minimise
microbial growth on the baffles and the fermenter wall.
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4. Aeration System (Sparger):

The device used to introduce air into the fermenter broth is called sparger. Spargers are
of the following three basic types: (1) porous spargers, (2) orifice spargers and (3) nozzle
spargers. Porous spargers may be made of sintered glass, ceramics or a metal.

They are used primarily on a laboratory scale in non-agitated vessels. The bubble size
from such spargers is always 10 to 100 limes larger than the pore size of the sparger.
These spargers have low air throughput because pressure drops across the sparger, and
the fine holes often become blocked by microbial growth.

Orifice spargers consist of perforated pipes arranged in various ways, e.g., the sparger
pipe forming a ring below the impeller. In most cases, air holes are drilled on the
underside of the pipe and the holes are arranged in the form of ring or cross.

It is desirable that the

holes are at least 6 mm in
diameter to avoid
clogging by microbial
growth. These spargers
(without agitation) have
been used to a limited
extent in yeast
manufacture, effluent
treatment and in air-lift fermenters used for single-cell protein (SCP) production. Nozle
sparger consists of an open or partially closed pipe. Most modern fermenters (laboratory
to production scale) have a single open or partially closed pipe as a sparger that is ideally
placed centrally below the impeller.

It provides a stream of air bubbles. The sparger should be as far below the impeller as
possible to avoid flooding of the impeller in a stream of air bubbles. These spargers cause
a lower pressure loss than the other spargers and they are not easily blocked.
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In small fermenters, a combined sparger-agitator may be used. In this case, the air is
introduced via a hollow agitator shaft, and it comes out through holes drilled in the disc
between the blades and connected to the base of the main shaft. This design gives a
good aeration in baffled vessels over a range of agitator speeds.

Temperature Regulation:

The fermenter must have an adequate provision for temperature control. Both microbial
activity and agitation will generate heat. If this heat generates a temperature that is
optimum for the fermentation process, then heat removal or addition may not be
required.

But in most cases, this may not be the case; in all such cases, either additional heating or
removal of the excess heat would be required. Temperature control may be considered at
laboratory scale, and pilot and production scales.

1. In laboratory scale fermentations, normally little heat is generated. Therefore, heat has
to be added to the system; this can be achieved in the following ways: (a) the fermenter
may be placed in thermostatically controlled bath, (b) internal heating coils may be used,
(c) water may be circulated through a heating jacket, or (d) a silicone healing jacket may
be used. The silicone jacket consists of two silicone rubber mats, and heating wires
between these mats. This jacket is wrapped around the fermenter and is held in place by
Velcro strips.

2. In case of larger fermenters beyond a certain size, excess heat is generated, and the
fermenter surface becomes inadequate for heat removal. The size at which fermenter
surface becomes inadequate for heat removal will depend on the fermentation process
and the ambient temperature at which fermentation is being carried out. In such cases,
internal coils have to be used to circulate cold water through them for removing the
excess heat.

The cooling surface area necessary for temperature control will depend mainly on the
following factors: (i) temperature of cooling water, (ii) the culture temperature, (iii) the
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type of microorganism, and (iv) the energy provided by stirring. The average cooling area
for a 55,000 l fermenter may be considered to be around 50-70 m2; if the cold water
temperature were 14C, the broth temperature would cool down to 30C from 120C in
2.5 to 4 hours without stirring. The cooling water consumed during bacterial fermentation
in a vessel of this size would range between 500 to 2,000 l h-1. Fungal fermentation,
however, may need 2,000 to 10,000 l cooling water per hour as they have a lower
optimum temperature for growth.

The heating/cooling requirements for a specific fermentation process can be accurately

estimated by taking into account the overall energy balance of the process.

Foam Control:

Foam is produced during most microbial fermentations. Foaming may occur either due to
a medium component, e.g., protein present in the medium, or due to some compound
produced by the microorganism. Proteins are present in corn-steep liquor, pharma media,
peanut meal, soybean meal, etc.

These proteins may denature at the air-broth interface and form a protein film that does
not rupture readily. Foaming can cause removal of cells from the medium; such cell wills
undergo autolysis and release more proteins into the medium. This, in turn, will further
stabilize the foam. Five different patterns of foaming are recognized; these are listed
below.

1. Foaming remains at a constant level throughout the fermentation. Initial foaming is due
to the medium, but later microbial activity contributes to it.

2. Foaming declines steadily in the initial stages, but remains constant thereafter. This
type of foaming is due to the medium.

3. The foaming increases after a slight initial fall, in this case, microbial activity is the
major cause of foaming.
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4. The foaming level increases with fermentation duration; such foaming pattern is solely
due to microbial activity.

5. A complex foaming pattern that combines features of two or more of the above
patterns.

Foaming may lead to several physical and biological problems. Some examples
of physical problems are as follows:

(1) The working volume of the fermenter may decrease due to a circulation of oxygen-
depleted gas bubbles in the system.

(2) The bubble size may also decrease, and

(3) The heat and mass transfer rates may also decline.

(4) Foaming may interfere with the functioning of sensing electrodes resulting in invalid
process data, and incorrect monitoring and control of pH, temperature, etc. The biological
problems of foaming include (1) deposition of cells in the upper parts of the fermenter, (2
problems of sterile operation as the air filter exits of the fermenter become wet, and (3)
increased risk of contamination. In addition, (4) there may be product loss due to
siphoning of the culture broth.

Whenever excessive foaming occurs, the following approaches may be used to

resolve the problem:

(1) A defined medium may be used to avoid foam formation. This may be combined with
modifications in physical parameters like pH, temperature, aeration and agitation. This
approach will be successful in such cases where medium is the main culprit, but will fail
whenever microbial activity is the main contributor.

(2) Often the foam may be unavoidable; in such case, antifoam should be used. This is
the most standard approach to combat foaming.
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(3) A mechanical foam breaker may also be used. Antifoams are surface active agents;
they reduce surface tension in the foams and destabilize protein film by the following
effects: (a) hydrophobic bridges between two surfaces, (b) displacement of the absorbed
protein, and (c) rapid spreading of the surface film. Ideal antifoam should have the
following properties.

1. It should disperse rapidly and act fast on existing foam.

2. It should be used at a low concentration.

3. It should prevent new foam formation for a long time.

4. It should not be used up or degraded by the microorganism.

5. It should be nontoxic (to the microorganism as well as animals, including humans).

6. It should not interfere with downstream processing.

7. It should not cause problems in effluent treatment.

8. It should be safe to handle.

9. It should be cheap.

10. It should not affect oxygen transfer.

11. It should be heat stable for heat sterilization.

Several compounds meet most of these requirements, and have been found to be
suitable for different fermentation processes; these compounds are as follows: alcohols
(stearyl and octyl decanol), esters, fatty acids and their derivatives (especially,
triglycerides like cottonseed oil, linseed oil, soybean oil, sunflower oil, etc.), silicones,
sulphonates, and miscellaneous compounds like oxaline, Alkaterge C, and polypropylene
glycol.
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Many of the antifoams are of low solubility; therefore, they are added with a carrier like
lard oil, liquid paraffin and castor oil. There carriers, however, may be metabolized, and
they may affect the fermentation process. Further, many antifoams would reduce oxygen
transfer by up to 50% when used at effective concentrations.

Antifoams are generally added when foaming occurs during fermentation. But foam
control in fermentation industry is still an empirical art. Therefore, the best method of
foam control for a particular process in one factory is not necessarily the best for the
same process in other factories. Further, the design and operating parameters of the
fermenters may affect the properties and the foams produced during the fermentation
process.

Types of Fermenters:

A variety of fermenters have been described in the literature, but few of them have
proved satisfactory for large scale aerobic fermentations. The most commonly used
fermenters are based on a stirred upright cylinder with sparger aeration (Fig. 14.3).

The volumes ranging from 1 l to several thousand I. A general description of the following
types of fermenter is given in the following sections: (1) stirred tank reactor, (2) airlift
fermenter, (3) tower fermenter and (4) bubble up fermenter.

Stirred Tank Fermenter:

These are glass (smaller vessels) or stainless steel (larger volumes) vessels of 1-1,000 1
or even 8,000 1 (Namalva cells grown for interferon; but in practice their maximum size is
20 1 since larger vessels are difficult to handle, autoclave and to agitate the culture
effectively).

Stirred tank reactor is the choice for many (more than 70%) though it is not the best.
Stirred tank reactors have the following functions: homogenization, suspension of solids,
dispersion of gas-liquid mixtures, aeration of liquid and heat exchange. The Stirred tank
reactor is provided with a baffle and a rotating stirrer is attached either at the top or at
the bottom of the bioreactor. The typical decision variables are: type, size, location and
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the number of impellers; sparger size and location. These determine the hydrodynamic
pattern in the reactor, which in turn influence mixing times, mass and heat transfer
coefficients, shear rates etc. The conventional fermentation is carried out in a batch
mode. Since stirred tank reactors are commonly used for batch processes with slight
modifications, these reactors are simple in design and easier to operate. Many of the
industrial bioprocesses even today are being carried out in batch reactors though
significant developments have taken place in the recent years in reactor design, the
industry, still prefers stirred tanks because in case of contamination or any other
substandard product formation the loss is minimal. The batch stirred tanks generally
suffer due to their low volumetric productivity. The downtimes are quite large and
unsteady state fermentation imposes stress to the microbial cultures due to nutritional
limitations. The fed batch mode adopted in the recent years eliminates this limitation.
The Stirred tank reactors offer excellent mixing and reasonably good mass transfer rates.
The cost of operation is lower and the reactors can be used with a variety of microbial
species. Since stirred tank reactor is commonly used in chemical industry the mixing
concepts are well developed. Stirred tank reactor with immobilized cells is not favored
generally due to attrition problems; however by separating the zone of mixing from the
zone of cell culturing one can successfully operate the system.

Continuous-Flow culture systems, a type of stirred tank reactors, are either of chemostat
or turbidostat type. In both the types, cultures begin as a batch culture. In a chemostat
type, inoculated cells grow to the maximum density when some nutrient, e.g., a vitamin,
becomes growth limiting.

Fresh medium is added after 24-48 hours of growth, at a constant rate (usually lower than
the maximum growth rate of culture) and at an equal rate the culture is withdrawn. When
the rate of growth equals the rate of cell withdrawal, the cultures are in a steady state,
and both the cell density and medium composition remain constant. One of the
constituents of the medium is used at a lower concentration to make it growth-limiting.
However, chemostat is the least efficient or controllable at the cells maximum growth
rate hence the steady-state growth rates in them are much lower than the maximum.
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In contrast, in a turbidostat cells grow to achieve a pre-decided density (measured as

turbidity using a photoelectric cell). At this point, a fixed volume of culture is withdrawn
and the same volume of fresh normal (not having a growth-limiting factor) medium is
added; this lowers the cell density or turbidity of the culture.

Cells keep growing, and once the culture reaches the preset density the fixed volume of
culture is replaced by fresh medium. This system works really well when the growth rate
of the culture is close to the maximum for the cell line.

The continuous-flow cultures provide a continuous source of cells, and are suitable for
product generation, e.g., for the production of viruses and interferons. It is often
necessary to use a two-stage system in which the first stage supports cell growth, while
the second stage promotes product generation.

Airlift Fermenter:

An airlift fermenter consists of a gas light baffled riser tube or draught tube (broth rises
through this tube) connected to a down-comer tube
(broth flows down through this tube). The riser tube
may be placed within the down-comer tube as shown
in Fig. 14.4, or it may be externally located and
connected to the latter (Fig. 14.5). Air/gas mixture is
introduced into the base of the riser tube by a
sparger.

The aerated medium/broth of the riser tube has a

lower density, while that in the down-flow tube it is
relatively much less aerated and, as a consequence, has a higher density. This density
difference drives the circulation of broth.

The lighter medium in the rise tube flows upward till it reaches the gas disengagement
space of the fermenter. The O2 is continuously consumed by the cells and CO2 is
generated by respiration.
 17

The bulk of CO2 and other gases move out of the

medium broth into the gas phase, and the un-aerated
medium flows down through the down-flow tube.
Circulation times in loops of 45 m height may be 120
seconds.

Single cell protein (SCP) production by Marlow Foods,

U.K uses an air-lift fermenter in which the riser tube is
externally placed (Fig. 14.6). Air and gaseous ammonia
are introduced at the base of riser tube, while
sterilized glucose, biotin and mineral salts are pumped into the fermenter at the base of
the down-flow tube.

An internal heat exchanger coil is located at the bottom loop connecting the riser and
down-flow tubes; it maintains the temperature at 30C. The upper loop connecting the
riser and down-flow tubes acts as an air outlet assembly through which CO2 is
continuously extracted. The removal of CO2 and continuous consumption of O2 dissolved
in the medium increases the density of the culture broth, which causes it to settle down
through the down-flow tube.

SCP is harvested through a port at the base of riser tube, which leads into an RNA
reduction vessel; steam is injected into the vessel to raise the temperature to 60C, which
reduces RNA content of SCP. After RNA reduction, SCP is harvested and processed.
 18

This air-lift fermenter of 43 m3 volume is

used in a continuous mode for the
production of mycoprotein Quorn from
Fusarium gaminareum grown on wheat
starch-based medium. It allows
production of long hyphae due to low
shear, which is the preferred form of the
product.

However, it gives lower biomass yields

(only 20 g l-1) due to lower oxygen
transfer rates in the high viscosity broth
resulting from fungal hyphae. This fermenter is a modification of that designed by ICI pic,
U.K. for SCP production using methanol as substrate.

The fermenter was developed to reduce production costs by minimising cooling costs
since agitated vessels would generate additional heat. ICI pic used it in a continuous
process to produce SCP for animal feed, but the process had to be discontinued because
of high methanol cost and competition from animal feeds based on protein-rich crop
produce. The mycoprotein production is, however, primarily for animal food.

Modifications of airlift fermenters include various modifications of draught (riser) tubes

and multiple air-lift fermenters.

(1) In one modification of draught tube, stainless steel four-mesh tubes were placed at
the top and bottom of the tube. This fermenter was used for growing Aspergillus terreus
for itaconic acid production. The sieves modulated the fungal morphology so that the
biomass was in an intermediate state between pellets and pulp. The type of culture gave
double the yields of itaconic acid per unit volume per unit culture time.
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(2) The multiple air-lift fermenters has three air-lift fermenters placed in a single vessel.
The medium is fed into the central fermenter from where it flows in the middle one and
then finally into the outer compartment from where it is eventually discharged.

(3) Another modification of air-lift fermenters is described in Section 14.9.3.4.

Animal cell cultures are also grown in such vessels that

are both aerated and agitated by air bubbles
introduced at the bottom of vessels (Fig. 14.7). The
vessel has an inner draft lube through which the air
bubbles and the aerated medium rise since aerated
medium is lighter than non-aerated one; this results in
mixing of the culture as well as aeration. The air
bubbles lift to the top of the medium and the air
passes out through an outlet.

The cells and the medium that lift out of the draft tube
move down outside the tube and are re-circulated.
O2 supply is quite efficient but scaling up presents
certain problems. Fermenters of 2-90 1 are
commercially available but 2,000 1 fermenters are being used for the production of
monclonal antibodies.

Tower Fermenter:

A tower fermenter has been defined by Green-shields and co-workers as an elongated

non-mechanically stirred fermenter that has an aspect ratio (height to diameter ratio) of
at least 6 : 1 for the tubular section and 10 ; 1 overall, and there is a unidirectional How of
gases through the fermenter. There are several different types of tower fermenters, which
are grouped as follows on the basis of their design: (1) bubble columns, (2) vertical-tower
beer fermenter and (3) multistage fermenter systems.
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1. Bubble Column Tower Fermenters:

These are the simplest type of tower fermenters; they consist of glass or metal tubes into
which air is introduced at the base. Fermenter volumes from 3 / to up to 950 / have been
used, and the aspect ratio may be up to 16 : 1. These tower fermenters have been used
for citric acid and tetracycline production, and for a range of other fermentations based
on mycelial fungi.

2. Vertical-Tower Beer Fermenters:

These fermenters were designed for beer production and to maximise yeast biomass
yields. A series of perforated plates are placed at intervals to maximise yeast yields. It
has a settling zone free of gas; in this zone, yeast cells settle down to the bottom and
return to the main body of the tower fermenter, and clear beer could be removed from
the fermenter. Tower of up to 20,000 / capacity and capable of producing up to 90,000 I
beer per day have been installed.

3. Multistage Tower Fermenters:

In these fermenters, a column forms the body of vessel, which is divided into
compartments by placing perforated plates across the fermenter. About 10% of the
horizontal area of plates is perforated. In a variant of this type of fermenter (down-flow
tower fermenter), the substrate is fed in at the top and overflowed through down spouts
to the next section, and the air is supplied from the base. These fermenters have been
used for continuous culture of E. coli, S. cerevisiae (bakers yeast), and activated sludge.

Bubble-up Fermenter:

It is a bubble column fermenter that is fitted with an internal cooling coil (Fig. 14.8). Air is
introduced from the bottom of the column. In this vessel, the cooling coil effectively
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separates the column into an inner riser/draught tube and

the outer down-flow tube. The cooling coil assembly
functions as a leaky draught tube.

The culture broth rises in the compartment enclosed by the

cooling coils and it moves down in the compartment outside
the coil, although back- mixing also occurs through the
coils. The region above the cooling coil shows good mixing,
and there were no poorly oxygenated zones in the vessel. It
can generate liquid velocities of 1 m sec-1, giving circulation
times of 9-12 seconds and mixing times of 14-18 seconds.

FLUIDISED BED BIOREACTOR

Fluidized bed bioreactors (FBB) have received increased attention in the recent years due
to their advantages over other types of reactors. Most of the FBBs developed for
biological systems involving cells as biocatalysts are three phase systems (solid, liquid &
gas). The fundamentals of three phase fluidization phenomena have been
comprehensively covered in chemical engineering literature. The FBBs are generally
operated in co-current upflow with liquid as continuous phase and other more unusual
configurations like the inverse three phase fluidized bed or gas solid fluidized bed are not
of much importance. Usually fluidization is obtained either by external liquid re-circulation
or by gas fed to the reactor. In the case of immobilized enzymes the usual situation is of
two-phase systems involving solid and liquid but the use of aerobic biocatalyst
necessitate introduction of gas (air) as the third phase. A differentiation between the
three phase fluidized bed and the airlift bioreactor would be made on the basis that the
latter have a physical internal arrangement (draft tube), which provides aerating and non-
aerating zones. The circulatory motion of the liquid is induced due to the draft tube.
Basically the particles used in FBBs can be of three different types: (i) inert core on which
the biomass is created by cell attachment. (ii) Porous particles in which the biocatalyst is
entrapped.(iii) Cell aggregates/ flocs (self-immobilization). In comparison to conventional
mechanically stirred reactors, FBBs provide a much lower attrition of solid particles. The
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biocatalyst concentration can significantly be higher

and washout limitations of free cell systems
can be overcome. In comparison to packed bed
reactors FBBs can be operated with smaller size
particles without the drawbacks of clogging, high
liquid pressure drop, channeling and bed compaction.
The smaller particle size facilitates higher mass
transfer rates and better mixing. The volumetric productivity attained in FBBs is usually
higher than in stirred tank and packed bed bioreactors. There are several successful
examples of using FBBs in bioprocess development.

PACKED BED BIOREACTOR

Packed bed or fixed bed bioreactors are commonly used with attached biofilms
especiallyin wastewater engineering. The use of packed bed reactors gained importance
after the potential of whole cell immobilization technique has been demonstrated. The
immobilized biocatalyst is packed in the column and fed with nutrients either from top or
from bottom. One of the disadvantages of packed beds is the changed flow characteristic
due to alterations in the bed porosity during operation. While working with soft gels like
alginates, carragenan etc the bed compaction
which generally occurs during fermentation
results in high pressure drop across the bed. In
many cases the bed compaction was so severe
that the gel integrity was severely hampered.
In addition channeling may occur due to
turbulence in the bed. Though packed beds
belong to the class of plug flow reactors in
which backmixing is absent in many of the packed beds slight amount of backmixing
occurs which changes the characteristics of fermentation. Packed beds arc generally used
where substrate inhibition governs the rate of reaction. The packed bed reactors are
widely used with immobilized cells. Several
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modifications such as tapered beds to reduce the pressure drop across the length of the
reactor, inclined bed, horizontal bed, rotary horizontal reactors have been tried with
limited success.

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