Shroff S.R.

Rotary Institute of Chemical Technology

(SRICT)
Principal Supporter & Sponsor-UPL Ltd / Shroff family
Managed By Ankleshwar Rotary Education Society
Approved by AICTE, New Delhi, Govt. of
Gujarat & GTU Affiliated

Fermentation
Syllabus
• Principles of surface and solid state fermentation,
• Design of different fermentors and the biochemical engineering
aspects.
• Process control of fermentations.
• Fermentation technology of industrial chemicals, organic acids, amino
acids, vitamins, polysaccharides, antibiotics, etc.
• Enzyme fermentation and technology including immobilization and
enzyme reactors.
• Fermentative animals, and other developments
• Fermentation is derived from the latin word fervere, meaning to boil.
• The term fermentation is used in biochemical sense to mean energy
generation process through organic compounds which act as electron
donors as well as terminal electron acceptors.
• Industrial microbiologists describe fermentation as any process for the
production of product by mass culture of a micro-organism.
• The first industrial process for production of a microbial metabolite,
alcohol, was by the action of yeast on malt or fruit extracts.
• Fermentation is a metabolic process that produces chemical changes in
organic substrates through the action of enzymes
• In microorganisms, fermentation is the primary means of producing ATP by
the degradation of organic nutrients anaerobically
• The science of fermentation is known as zymology.
 Components of a Fermentation Process:
• Regardless of the type of fermentation the process is divided into six basic
parts (except transformation processes where microbial cells are used to
convert compounds to structurally related more valuable compounds ):

1. The formulation of media to be used in culturing the process organism

during the development of the inoculum and in the production
fermenter
2. The sterlization of the medium, fermenters and ancillary equipment
3. The production of an active, pure culture in sufficient quantity to
inoculate the production vessel
4. The growth of the organism in the production fermenter under
optimum conditions for product formation
5. The extraction of the product and its purification
6. The disposal of effluents produced by the process
 Biomass
Production Fermenter

Culture
fluid Cell
Stock c ulture Separation
S hak er Flask
Se ed Fermenter
Cell-free
supernatant
Medium sterlisation

Medium formulation Product

extraction

Medium raw materials Product

Effluent
Purification treatment

TYPICAL FERMENTATION PROCESS

Product
Packaging
• Major types of fermentations:
i. Those that produce microbial cells (biomass) as the product
ii. Those that produce microbial enzymes
iii.Those that produce microbial metabolites
iv. Those that produce recombinant product(artificially produced & often purified
product)
v. Those that modify a compound which is added to the fermentation – the
transformation process
• Microbial biomass
• Two major process
• Production of Yeast for the baking industry
• Production of microbial cells as human / animal food
• Microbial Enzymes
• Produced from-
• Animals
• Plants
• Microbes
 • Microbial system for enzymes offers:
• Ease of production
• Advantage of mass production by well established fermentation techniques
• Enzymes of animal origin can be synthesized with microorganisms (recombinant technology)
• Easy to control and modify
• Microbial metabolites:
• There are a number of stages in the growth of microbial culture
• Lag phase: On inoculation of a culture into a nutrient medium growth does not appear to occur.
This phase is called the time of adaptation.
• Log or Exponential Phase: The cells grow at a constant maximum rate after the lag phase. During
this phase the products produced are essential for the growth of the cells such as proteins, lipids ,
carbohydrates, amino acids, nucleotides, etc. These primary products of metabolism. The phase is
also referred as Trophophase. Products of commercial importance are produced by fermentation
such as citric acid, ethanol, vitamins, polysaccharides, lysines, glutamic acid, etc
• Death Phase: The period in which the viable cell numbers declines. In this phase, secondary
metabolites which do not have any function in cell metabolism. The phase under which these
secondary metabolites are produced is referred as idiophase
• The primary and the secondary metabolites are interrelated in the sense that the secondary
metabolites are produced from the products of primary metabolites. Many secondary
metabolites are antimicrobial, enzyme inhibitors, growth promotors and pharmacological
active.
• Recombinant Products:
• A large number of potential fermentation products are possible by using the
recombinant technology. The genes of higher organisms are introduced in
microbes such as Escherichia Coli, Saccharomyces cerevisiae and the
filamentus fungi to produce products such as insulin, interferon, human
serum albumin, epidermal growth factor, calf chymosin and bovine
somatostatin.
• Transformation Processes:
• Microbial cells are used to convert compounds to structurally related more
valuable compounds. This is done by taking advantage of the fact that they
behave as chiral catalysts where high positional specificity and
stereospecificity are shown in their transformations. Microbial process offer
low temperature and pressure transformations and are essentially non-
polluting. Production of vinegar (conversion of ethanol to acetic acid) is a fine
example of this process. Other high value products include prostaglandins,
steroids and antibiotics.
• Basic functions of a Fermenter for microbial and animal cell culture
• The main function of a fermenter is to provide a controlled environment for
the growth of microorganisms or animal cells, to obtain a desired product.
• Points to be considered in designing and constructing a fermenter:
1. The vessel should be capable of being operated aseptically for a number of days and
should be reliable in long term operations and meet the requirements of container
regulations
2. Adequate aeration and agitation should be provided to meet the metabolic
requirements of the micro-organism. However, mixing should not cause any damage
to the organism.
3. Power consumption should be as low as possible
4. A system of temperature should be provided
5. A system of pH control should be provided
6. Sampling facilities should be provided
7. Evaporation losses from the fermenter should not be excessive
8. The vessel should be designed to require the minimal use of labour in operation,
harvesting, cleaning and maintenance.
 9. Ideally the vessel should be suitable for a wide 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 instead of flange joints whenever possible.
11. The vessel should be of similar geometry to both smaller and larger vessels
in pilot plant or plant to facilitate scale-up.
12. The cheapest material which enable satisfactory results to be achieved
should be used.
13. There should be adequate service provisions for individual plants
• The most commonly used fermenters are based on stirred upright
cylinders having single or multiple impellers with sparger aeration.
• A very small percentage of the different types of fermenters have
been satisfactory for industrial aerobic fermentations
• Utilities or service provisions for a fermentation plant:
1. Compressed air
2. Sterile compressed air(1.5 - 3 atm)
3. Chilled water (12-15oC)
4. Cold water (4oC)
5. Hot water
6. High Pressure Steam
7. Steam condensate
8. Electricity
9. Stand-by-generator
10. Drainage of effluents
11. Motors
12. Storage facilities for media components
13. Control and monitoring equipment for fermenters
14. Maintenance facilities
15. Extraction and recovery equipment
16. Accessibility for delivery of material
17. Appropriate containment facilities
• Aseptic operation and containment:
• Aseptic operation involves protection against contamination.
• Containment involves prevention of escape of viable cells from the fermenter or
downstream equipment.
• To establish the appropriate level of containment risk assessment is to be carried
out. This is done through categorization of process microorganisms and designation
of its appropriate level of containment at research or industry sites say as per the
European Federation of Biotechnology (see fig)
• In the non-genetically engineered organisms placed in Hazard Grp-I require only Good
Industrial Large Scale Practice(GILSP). The processes in this category need to be operated
aseptically but no containment is necessary including prevention of organisms, whereas those
placed in Hazard Grp-4 stringent level-3 requirements are to be met.
• Genetically engineered organisms are classified as either harmless (Grp-I) or potentially
harmful(Grp-II). The process is further classified as small scale (A) or large scale (B). Large
scale fall in two categories IB or IIB. IB processes require containment level B1 and are subject
to GILSP. Whereas IIB processes are further assessed to determine the most suitable
containment level, ranging from B2 to B4 corresponding to levels 1 to 3 for non-genetically
engineered organisms.
 Process Organism

Genetically engineered
Non-genetically engineered

Hazard Group Allocation Harmless Potentially Harmful

Group-I Group-II

1,2,3,4 Small Large Small Large

Scale Scale scale Scale

A B A B
Containment Level
Containment
level
GILSP 1 2 3 B1 B2 B3 B4
GILSP
 • Most micro-organisms used in industrial processes are in the lowest hazard group and require only
GILSP barring a few especially those used in viral vaccine production
• Body Construction:
• Construction material:
• In fermentations with strict aseptic requirements it is important to select materials
that can withstand repeated steam sterilization cycles. On a small scale (1-30dm3)
glass and stainless steel can be used.
• Advantages of using Glass:
• Smooth surfaces
• Non-toxic
• Corrosion proof
• Easy to examine the interiors
• Two basic types of fermenter are used:
• A glass vessel with a round or flat bottom and a top flanged carrying plate. This type have to
be sterilized by autoclaving. The largest practical diameter for glass fermenters is 60cm
• A glass cylinder with stainless steel top and bottom plates. These fermenters may be sterilized
insitu but 30cm diameter is the upper size limit to safely withstand working pressures. Vessels
with two SS plates cost twice as that one with a single plate.
• Pilot and industrial scale are normally constructed of SS or at least have a SS
cladding to limit corrosion. As per American Iron and Steel Institute(AISI) steels
containing >4% chromium are classified as SS and those <4% as steel alloys.
• The corrosion resistance of SS is due to a thin film of hydrous oxide film on the
surface of the metal. The composition of this film varies with different steel alloys
and is stabilised with chromium which is considered to be continuous, non-
porous, insoluble and self healing.
• Increasing the chromium content enhances resistance to corrosion, but only steel
containing at least 10-13% chromium develop an effective film.
• The inclusion of nickel in high percent chromium steels enhances their resistance
and improves their engineering properties.
• The presence of Molybdenum improves the resistance of stainless steel to
solutions of halogen salts and pitting by chloride ions in brine or sea water.
• Corrosion resistance can be improved by tungsten, silicon and other elements
• Mild steel coated with glass or phenolic epoxy materials have also been used
• AISI grade 316 steel is commonly used in fermenter construction. This
contains:
• 18% chromium
• 10% nickel
• 2-2.5% molybdenum
• AISI grade 304 is extensively used in brewing equipment and it
contains
• 18.5% chromium
• 10% nickel
• The thickness of the construction material will increase with scale. At
300000 to 400000 dm3 capacity, 7mm plate may be used for the side
of the vessel and 10mm plate for the top ad bottom, which should be
hemispherical to withstand pressures
• Temperature Control:
• Heat is generated in the reactor by:
• Microbial action
• Mechanical agitation
• Heat may have to be given or removed depending upon the process. This is
done using cooling through jacket and/or internal coils.
• It is impossible to specify accurately the necessary cooling surface of a
fermenter since the temperature of the cooling water, the sterilization
process, the cultivation temperature, the type of micro-organism and the
energy supplied by stirring can vary considerably in different processes.
• A cooling area of 50-70m2 may be taken as average for 55000 dm3 fermenter
and with a coolant temperature of 14oC the fermenter may be cooled from
120 to 30oC in 2.5 – 4hrs without stirring. The consumption of cooling water
in this size of vessel during a bacterial fermentation ranges from 200 to
2000dm3 h-1 while fungi might require 2000 to 10000dm3 h-1 due to lower
optimum temp for growth.
• To accurately estimate heating and cooling requirements for a specific
process it is important to consider all the contributing factors. The over all
energy balance of a fermenter during normal operation can be estimated
through the following equation:
Q met + Q ag + Qgas = Q acc + Q exch + Q evap + Qsen

• Where Qmet = rate of heat generation due to microbial metabolism

• Qag = rate of heat generation due to mechanical agitation
• Qgas = rate of heat generation due to aeration power input
• Qacc = Rate of heat accumulation by the system
• Qexch = rate of heat transfer to the surroundings and/or heat exchanger
• Qevap = rate of heat loss by evaporation
• Qsen = Rate of enthalpy gain by the flow streams
• The equation can be rewritten as:
Q exch = Qmet + Qag + Q gas - Qacc - Q evap - Qsen

• The cooling requirements (jackets and / or pipes ) to remove the

excess heat from a fermenter may be determined by the formula:

Watts per squared meter kelvin

• Aeration and Agitation
• Aeration is done to provide microorganism with sufficient oxygen to carry
out metabolic requirements.
• The type of aeration-agitation system used depends on the characteristics
of the fermentation process under consideration
• The bubble aerators without mechanical agitators have the advantage of
lower equipment and power costs. In this process broths of low viscosity
and low total solids are used.
• Mechanical agitation is usually required in fungal and actinomycete (a
heterogenous gram positive bacteria) fermentation.
• The structural components of the fermenter involved in aeration and
agitation are:
• The agitator(impeller)
• Stirrer glands and bearing
• Baffles
• The aeration system(sparger)
• Agitator
• Agitator is required to achieve:
• Bulk fluid and gas-phase mixing
• Air dispersion
• Oxygen transfer
• Heat transfer
• Suspension of solid particles
• Maintaining a uniform environment in the vessel

• Types of Agitators:
• Agitators may be classified as
• (a)Disc turbine
• (b)Vaned discs,
• (c)Open turbines of variable pitch
• (d)Propellors
Different Impeller Types. (a)
Marine-type propellers; (b) Flat-
blade turbine, Wi = Di/5. © Disk
flat-blade turbine, Wi = Di/5, Di =
2Dt/3, Li = Di/4; (d) Curved-blade
turbine, Wi = Di/3; (e) Pitched-
blade turbine, Wi = Di/8; and (f)
Shrouded turbine, Wi = Di/8.

Dt = tank diameter,
Di = impeller diameter
Wi = impeller blade height
• Disc turbine: It consists of a disc with a series of rectangular vanes set in a vertical
plane around the circumference. Disc turbine is the most suitable in a fermenter
since it can break up a fast air stream without itself becoming flooded in air
bubbles.
• Vaned Disc: It 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.
• Variable Pitch open turbine & Marine propeller: The vanes are attached to a
boss on the agitator shaft. The air bubbles do not hit any surface before
dispersion by the vanes or blades
NEW AGITATORS

Scaba Agitator

Lightnin A315 Agitator

Prochem Maxflo T agitator

• Scaba 6SRTG agitator
• Developed to handle problems associated with efficient blending in high viscosity
fermentations.
• At a given power input can handle a high air flow rate before flooding.
• Radial-flow agitation is better for bulk blending than a Rushton turbine, but does not
give good top to bottom blending in a large fermenter which leads to lower
concentrations of oxygen in broth away from the agitators and higher concentrations
of nutrients, acid or alkali or anti-foam near to the feed points.
• Prochem Maxflo Agitator
• Consists of 4-6 hydrofoil blades set at a critical angle on the central hollow hub.
• A hydrodynamic thrust is created during rotation increasing the downward pumping
capacity of the blades.
• This design minimizes the drag forces(is a forceacting opposite to the relative motion of any object
moving with respect to a surrounding fluid.) associated with the rotation of the agitator such
that the energy losses due to drag are low resulting in low power number
• BAFFLES
• Baffles are metal strips that prevent vortex formation around the walls of the
vessel.
• These metal strips attached radially to the wall for every 1/10 th of vessel
diameter.
• Usually 4 baffles are present but when the vessel diameter is over 3dm3
around 6-8 baffles are used.
• There should be enough gap between wall and baffle so that scouring
action(removing loose particle from interior surfaces and flushing out) around vessel is facilitated. This
movement minimizes microbial growth on baffles and fermentation walls. If
needed cooling coils may be attached to baffles.
• STIRRER GLANDS AND BEARINGS
• The entry point of stirrer into fermenter may be from top to bottom or sides.
Mostly used from bottom so that that leaves more space for entry ports on top.
There are four types of stirrer glands and bearings.
• 1) Stuffing box
• a. Sealed by several layers of packing rings of asbestos or cotton yarn-pressed against the
shaft by a gland follower
• b. At high speeds- packing wears – pressure should be applied to ensure tightness
• c. Difficult to sterilize- satisfactory heat penetration
• d. Sufficient for GILSP(good industrial large scale practice) containment
• 2) Mechanical seal
• a. 2 parts; i) stationary in the bearing housing, ii) other rotates on the shaft.
• b. Two parts pressed together by springs or expanding bellows
• c. Steam condensate use to lubricate and cool seals
• d. safe for containment
• e. double mechanical seal for level 2
• f. at level 2 and 3, the condensate is piped to a kill tank
• g. Disinfectants flushed through the seal
• h. steam condensate outlet monitoring indicates any seal failure
• 3) Magnetic drives (some animal cell cultures)
• a. shaft does not pierce the vessel
• b. two magnets- one driving, held in bearing in housing on outside of head
plate and one driven, placed on one end of impeller shaft held in bearing in
suitable housing
• c. ceramic magnets –magnetic power cross 16mm gap
• d. 300 – 2000 rpm rotation possible
• 4) Simple bush seals
• Disadvantage of double seals are more difficult to assemble, difficult to detect
failure of seal from normal and dead spaces and seals leading to
contamination. Hence simple bush seal is preferred in some cases.
• AERATION SYSTEM (SPARGER)
• Sparger is a device for introducing air into fermenter. Aeration provides
sufficient oxygen for organism in the fermenter. Fine bubble aerators must
be used. Large bubbles will have less surface area than smaller bubbles
which will facilitate oxygen transfer to a greater extent. Agitation is not
required when aeration provides enough agitation which is the case Air lift
fermenter. But this is possible with only for medium with low viscosity and
low total solids. For aeration to provide agitation the vessel
height/diameter ratio (aspect ratio) should be 5:1. Air supply to sparger
should be supplied through filter. There are three types of sparger viz.
porous sparger, orifice sparger and nozzle sparger.
• 1. Porous sparger: made of sintered glass, ceramics or metal. It is used only
in lab scale-non agitated vessel. The size of the bubble formed is 10-100
times larger than pore size. There is a pressure drop across the sparger and
the holes tend to be blocked by growth which is the limitation of porous
sparger.
• 2. Orifice sparger: used in small stirred fermenter. It is a perforated
pipe kept below the impeller in the form of crosses or rings. The size
should be ~ ¾ of impeller diameter. Air holes drilled on the under
surfaces of the tubes and the holes should be atleast 6mm diameter.
This type of sparger is used mostly with agitation. It is also used with
out agitation in some cases like yeast manufacture, effluent
treatment and production of SCP.
• 3. Nozzle sparger: Mostly used in large scale. It is single open/partially
closed pipe positioned centrally below the impeller. When air is
passed through this pipe there is lower pressure loss and does not get
blocked.
• 4. Combined sparger agitator: This is air supply via hallow agitator
shaft. The air is emitted through holes in the disc between the blades
of agitator. The design gives good aeration in baffled vessels.
The achievement and maintenance of
aseptic conditions
• The following operations have to be performed according to
specifications to maintain aseptic conditions and containment during
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 and other supplements
5. Sampling
6. Foam control
7. Monitoring and control of various parameters
• On a small scale <10dm3 the biohazard risk can be controlled by a combination of
containment cabinets and work practices.
• When the volume of culture is > 10dm3, GILSP is required for non-pathogenic and
non-toxigenic agents
• For level 1, B2 or higher containment levels the following points need to be
considered when designing a fermenter
• All vessels containing live organism should be suitable for steam sterilization and have sterile
vent filters
• Exhaust gas should pass through sterile vent filters
• Seal on flange joint shall be fitted with a single ‘O’ ring at the lower levels of containment
• For containment level B3 or B3/4 double ‘O’ ring or double ‘O’ ring with a steam barrier
• Suitable seals should be provided at the entry ports for sensor probes, inoculum, sampling,
medium addition, acid , alkali and antifoam
• Rotating shafts into a closed system should be sealed with a double acting mechanical seal
with steam or condensates between the seals
• During operation a steam barrier should be maintained in all fixed piping leading to the
‘contained’ vessels
• Appropriate pressure release facilities to be provided
• Sterilization of fermenter
• Fermenter to be designed so that it can be steam sterilized under pressure
• The medium may be sterilized in the vessel or separately, and added to the
vessel aseptically
• If the medium is sterilized in situ then its temperature must be raised before
injection of the steam to prevent the formation of large condensates.
• Every point of entry and exit to the fermenter is a potential source of
contamination and hence steam should be introduced at such points
• All pipes should be constructed simply and should slope towards drainage
points.
• Each drainage point should be fitted with a steam trap
• Fermenter should be free of crevices (narrow opening) of 0.05mm depth
especially when using animal cells
• Welded joints should be used wherever possible
• Sterilization of air supply
• Air is sterilized through two processes: Heat and Filtration
• Sterilization through heat for full system is too costly
• Glass wool, Glass fibre, mineral slag wool have been used for
filtration. These have been replaced with cartridge type filters in
fermenters.
• The filter also needs to be sterilised in association with the
fermenter
• Sterilization of the exhaust gas from the fermenter
• Sterilisation of the exhaust is achieved by passing through a 0.2micron filter
put on the outlet pipe.
• For satisfactory operation a cyclone separator and a coalescer (A coalescer is a
technological device performing coalescence. They are primarily used to separate emulsions into their components via
various processes; operating in reverse to an emulsifier. )is included upstream of two filters in
series
• Addition of inoculum, nutrients and other supplements
• When operating under GILSP both the addition vessel and the fermenter is
placed under positive pressure and the addition port is equipped with a
steam supply.
• For containment levels 1 and B2, the addition is carried out in such a way that
the release of microorganism is restricted. This is done by aseptic piercing of
membranes or connections with steam locks
• At containment level 2 & B3/4 no microorganism must be released during
inoculation or other additions. This is done by tight screwing and clamping
and making all pipelines steam sterilizable
• Sampling:
• Sampling points are fitted to large fermenters so as to maintain sterility.
• 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.

Sampling port
• Feed Ports:
• Addition of nutrients, acid /alkali to fermenters are normally made via silicon
tubes which are autoclaved and pumped by a peristaltic pump(a series of
wave like movement-a diaphragm pump) after aseptic connections
• In large fermenters, the nutrient reservoir and associated piping are usually
an integrated part which can be sterilized with the vessel
• Sensor Probes:
• Double ‘O’ ring seals have been used to provide aseptic seals for glass
electrodes in stainless steel housing in fermenters using GILSP. System is
suitable for level 1 & B2 type of containment
• For containment levels 2 & B3/4 probes are fitted with triple ‘O’ rings 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 danger of leakage of broth
• Foam control:
• It is important to minimize foam in any fermentation process
• With excessive foam there is a danger of filters getting wet resulting in
contamination
• Siphoning may also result due to foam resulting in loss of part or entire contents of
the fermenter.
• Antifoam agents, mechanical foam breakers are used to overcome problems
associated with foam.
• Commonly used agents are insoluble oils, polydimethylsiloxanes and other silicones,
certain alcohols, stearates and glycols
• Valves and Steam Traps
• Valves attached to fermenters are used to control the flow of gas and liquid.
• The valves may be
• Simple ON/Off valve which are either fully open or closed
• Valves which provide coarse control of flow rates
• Adjustable valves which may be used for accurate control of flow rates
• Safety valve which allow unidirectional flow
• Gate Valve:
• This is a general purpose valve used in steam or water line for fully closing
and opening. This is not used as a controlling valve
• A sliding disc is moved in or out of the flow path by turning the stem of the
valve
• The flow path is such that the pressure drop is minimal.
• Unsuitable for aseptic conditions
• Globe Valves:
• A horizontal disc or plug is raised or lowered in its seating to control the rate
of flow.
• This type of valve is very commonly used for regulating the flow of water or
steam since it may be adjusted rapidly
• There is high pressure drop in the flow path.
• Unsuitable for aseptic conditions
Piston valve
• Piston valve:
• It is similar to globe valve except that the flow is controlled by a piston passing
between two packing rings
• The design is efficient for aseptic operations
• The pressure drop is similar to globe valves
• Steam is passed to sterilize the part open valve
• Needle Valve:
• This is also similar to globe valve except that the disc is replaced with a tapered plug
or a needle fitting into tapered valve seat
• It has limited aseptic applications
• The valve can give a fine controlled steam or liquid flow.
• Plug valve:
• It provides a good flow control
• In this valve there is a parallel or tapered plug sitting in a housing through which an
orifice has been machined
• This type of valve has a tendency to leak or sieze up, but can be controlled with the
use of lubricants
• Ball Valve:
• The valve element is a stainless steel valve through which an orifice is
machined.
• The ball is sealed between two wiping surfaces which wipe the surface and
prevent the deposition of the mater at this point.
• The orifice tin the ball is of the same diameter of the pipeline giving excellent
flow path.
• The valve is suitable for aseptic operations and can be operated at high
temperatures & pressures.
• Butterfly valve:
• It consists of a disc which rotates about a shaft in a housing. The disc closes
against the seal to stop flow of the liquid
• It is used in large diameter pipes operating under low pressure where
absolute closure is not essential.
• It is unsuitable for aseptic operations
Plug valve
• Pinch Valves:
• In this valve, a flexible sleeve is closed by a pair of pinch bars.
• The flow rate can be controlled 10 – 95% of rated flow capacity.
• It is suitable for aseptic operations with fermentation broths as there are no dead
spaces in the valve structure, and the closing mechanism is isolated from the
contents of the piping.
• Diaphragm Valve:
• The valve makes use of the flexible closure with or without a weir.
• Suitable for aseptic operations provided the diaphragm material can withstand the
sterilization operations
• The valve can be sued for on/off flow regulation, and for steam services with in
pressure limits
• Diaphragm failure is the primary fault of the valve.
• EPDM – Ethylene Propylene Diene Modified is the preferred material.
• A diaphragm valve with a steam seal on the clean side is considered a potentially
safer valve
Questions
• What is fermentation?
• With the help of a neat diagram show the basic features of a fermenter?
• What are the basic components of a fermentation process?
• List the major types of fermentations and briefly describe each one of them?
• What is the main function of a fermenter? List the main points in designing and constructing a fermenter?
• List utilities or service provisions for a typical fermentation plant?
• What is containment? Discuss how the containment levels are designated based on the process microorganism?
• With respect to fermentation, discuss:
• material of construction
• temperature control
• Write a note on agitation in a typical fermentation process?
• Write a note on aeration system(Sparger) in a typical fermentation process?
• Describe the major types of stirrer glands and bearings in a fermenter?
• Describe with the help of a diagram how air supplied to a fermenter is sterilized?
• Describe with the help of a diagram the working principle of a sampling port in a fermenter?
• Write a note on any two types of valves used in a fermenter?
• TYPES OF FERMENTERS
• The main function of a fermenter is to provide a controlled environment for the growth
of microorganisms or animal cells, to obtain a desired product. Few of the bioreactor
types are discussed below:
• STIRRED TANK FERMENTER
• Stirred tank reactor is the choice for many (more than 70%) though it is not the best.
Stirred tank reactor’s 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 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 reactor’s 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.