Recap

 Describe the nature of and explain the role of chemical engineers

in biochemical engineering.
 Define Biotechnology. Bioreactors, and Bioprocesses
 Explain some basic concepts of biology.
 Describe and explain the principles of enzyme kinetics (Michaelis-
Menten equation for Normal & Rapid equilibrium approach;
Integral MM equation etc)
 Identify the parameters and variables that affect microbial
growths (pH, temperature, concentration, time, radiation).
 Express kinetics of enzymatic reaction using mathematical
concepts. (Lineweaver-Burk method; Eadie-Hofstee method;
Hanes-Woolf method)
 Immobilized enzyme technology (Chemical & Physical methods) :
Non-covalent adsorption, Covalent attachment, entrapment in
crosslinked polymer, confinement in semipermeable membrane)
 Scope & outcomes
Scope: Analyze, design, and operate bioreactors

Outcomes:

1.Batch, Feb batch, & Continuous Bioreactors knowhow

2. Bioreactor Aeration and Agitation

3. Bioreactor Mixing

4. Bioreactor Scale-up operations

 Bioreactors
•Bioreactors: There are 2 main classes of bioreactors---
Anaerobic and Aerobic.
•In anaerobic bioreactor, microorganisms grow and create
product in the absence or near-absence of oxygen.
Examples of anaerobic fermenters include most fuel
processes (ethanol or isobutanol) and manufacture of some
organic acids.
•Aerobic fermentation takes place in the presence of oxygen,
which the organisms consume in order to grow and/or make
the desired product.
E. g. production of yeast, antibiotics, enzymes, and amino
acids
 Bioreactors
• The sizes of the bioreactor can vary over several orders of
magnitudes.
• Examples of bioreactors
• The microbial cell (few mm3),
• shake flask ( 100-1000 ml),
• laboratory fermenter ( 1 – 50 L),
• pilot scale (0.3 – 10 m3)
• plant scale ( 2 – 500 m3)
Bioreactor
 Requirements of a Bioreactor
(a) The design and construction of biochemical reactors must preclude foreign
contamination (sterility). Furthermore, aseptic conditions should be maintained
during the fermentation and ensure containment.
(b) Optimal mixing with low, uniform shear,
(c) Adequate mass transfer (oxygen)– Aerobic reactors
(d) Clearly defined flow conditions
(e) Feeding of substrate : prevention of under or overdosing
(f) Suspension of solids
(g) Gentle heat transfer
(h) Compliance with design requirements such as: ability to be sterilized; simple
construction; simple measuring, control, regulating techniques; scale up;
flexibility; long term stability; compatibility with up- downstream processes;
antifoaming measures.
 Configurations requirement- Bioreactor
 Heat and oxygen transfer configuration
· Sterilization procedures
· Foam control
· Fast and thorough cleaning system
· Proper monitoring and control system
 Traditional design is open cylindrical or rectangular vessels made from
wood or stone.
 Most fermentations are now performed in close system to avoid
contamination.

 It should be constructed from non-toxic, corrosion-resistant materials.

Small fermentation vessels of a few liters capacity are constructed from
glass and/or stainless steel.
Classification of Bioreactors on the
mode of operation

Batch bioreactor
Fed batch bioreactor
Continuous bioreactor
 Batch Bioreactor
 Batch bioreactors are simplest type of mode of reactor
operation. In this mode, the reactor is filled with medium and
the fermentation is allowed to proceed. When the
fermentation has finished the contents are emptied for
downstream  processing. The reactor is then cleaned, re-
filled, re-inoculated and the fermentation process starts
again.

 Molar or mass reactions are restricted in a variable

temperatures, stirring and PH. However the inflow and out
flow of the reaction products will remain constant
Batch Bioreactor
Continuous Bioreactor
 Continuous Bioreactor
 Here fresh media is continuously added and bioreactor fluid
is continuously removed
 As a result, cells continuously receive fresh medium and
products and waste products and cells are continuously
removed for processing
 The reactor can be operated for long periods of time without
having to be shut down.
 Both addition and removal of substrate and products are
done at the same rate so that the working volume remains
Constant.
 Growth rate of the bacteria in the reactor can be more
easily controlled and optimized
 In addition, cells can also be immobilized in continuous
reactors to prevent their removal which results in increased
productivity.
Feb batch Bioreactor
 Fed batch bioreactor
 It is the most common type of reactor used in industry.
 Here, fresh media is continuously or sometimes periodically
added to the bioreactor but unlike a continuous reactor,
there is no continuous removal.
 The Reactor is emptied or partially emptied when reactor is
full or fermentation is finished.
 Here, it is possible to achieve high productivities due to the
fact that the growth rate of the cells can be optimized by
controlling the flow rate of the feed entering the reactor.
Important fermentation products
 Key issues in Bioreactor design &
operation
A Bioreactor should comply with
Good manufacturing practice (GMP),
regulations
3 Qs: Design qualifications (DQ), Installation
qualifications, Operation qualifications. When 3
Qs are passed, it leads to Factory Acceptance
Test (FAT)
Validation: Established Documented Evidence
cGMP: This is employed strictly in drug
industry
 Key issues in Bioreactor design &
operation
• To accomplish the key issues, a Chemical Engineer must consider
the following areas:
 Suitable reactor parameters for biological, chemical, physical
systems
 Suitable Bioreaction parameters:
 Controlled temperature
 Optimum pH
 Sufficient substrate
 H2O availability
 Salts for nutrition
 Vitamins
 Oxygen
 Gas evolution
 Product & by-product removal
 Other parameters: Microbiology, sterilization, rheology, mixing,
agitator design, fluidization, heat/mass transfer, transport
phenomena, kinetics, instrumentation, process control
 Standards used in Bioreactor Design
consideration
• The standards used for Bioreactor design
considerations are:
 ASME-BPE-2000: American Standards for Bioprocess
Pressure Vessels
 GAMP for automation (Good Automated Manufacturing
Practice)
 ISO for qualification (International Organization for
Standardization)
 EN-46001 for medical devices
 Electrical guidelines
 Bioreactor Aeration & agitation
• Aeration refers to the process of introducing air to
increase oxygen concentration in liquids
• The aeration and agitation of the fermentation medium,
provides necessary oxygen to the industrial
fermentation process.
• Aeration may be performed by bubbling air through the
liquid, spraying the liquid into the air, or agitation of the
liquid to increase to increase surface absorption

• Agitation refers to uniform suspension of microbial

cells in homogeneous nutrient medium

• Structural components involved for aeration and

agitation: Impellers (agitator), baffles, Sparger
 Bioreactor Aeration & agitation

Major resistances for transfer of O2 from gas bubble to

cell
 Mass transfer steps during transfer of
O2 to cells
• Transfer from the interior of the gas bubble to the gas-liquid
interface
• Movement across the gas-liquid interface
• Diffusion through the relatively stagnant liquid film surrounding
the bubble
• Transport through the bulk liquid
• Diffusion through the relatively stagnant liquid film surrounding
the cells
• Movement across the liquid-cell interface
• If the cells are a floc, clump or solid particle – Diffusion through
the solid to the individual cell
• Transport through the cell wall, membrane and cytoplasm to the
site of reaction
 Bioreactor aeration and agitation
• Parts of the Bioreactor involved in Aeration and Agitation are:
 Impeller (Agitator)
 Sparger (for Aeration)

• Agitator: The main aim of the Agitator is to provide homogenous

environment all over the fermenter. It is also used for mixing of
different phases, and for oxygen and heat transfer.
Agitators
 Bioreactor aeration
• Spargers: A sparger is a tool used for introducing air into the
fermentation medium. Three basic types of sparger are used:
 The Porous Sparger,
 The Orifice Sparger (a perforated pipe) and
 The Nozzle Sparger (an open or partially closed pipe).
 Bioreactor aeration
 The Porous Sparger: The porous sparger is mainly used for
laboratory scale non-agitated fermentor. It is made up of
sintered glass, ceramics or metal.
 Bioreactor aeration
 The Orifice Sparger: In small stirred bioreactors the perforated
pipes are arranged below the impeller in the form of crosses or
rings, approximately three-quarters of the impeller diameter.
 Orifice or ring sparger is made of a perforated metal tube, a
porous ceramic material in the form of a ring.
 Bioreactor aeration
 The Nozzle Sparger: Most modern mechanically stirred
fermenter designs from laboratory to industrial scale have a
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. The
single-nozzle sparger causes a lower pressure loss than any other
sparger and normally does not get blocked.
 Bioreactor scale up operations
• What is Scale up operation: Scale up means reproducing in plant
scale equipment the results from a successful fermentation
made in a laboratory or pilot-scale equipment
• Scale up operations directly influences the production capacity
and efficiency of a bioprocess.

• Bioreactors are usually developed in 3 stages or scales:

 Bench or laboratory scale, where basic screening procedures are
carried out
 Pilot plant, where optimal operating conditions are ascertained.
 Plant scale, where the process is brought to economic fruition.
 Criteria of scale up
• During scale up operations, physical characteristics such as Mixing
time, shear, and mass transfer aspects are coupled, whereas, pH and
heat transfer are considered separately.
• Mass transfer takes places throughout the bioreactor
• Heat transfer occurs only at the boundary surfaces of heat exchange.
• Oxygen transfer is often the rate-limiting step in the aerobic
bioprocess due to the low solubility of oxygen in the medium. The
correct measurement and/or prediction of the volumetric mass
transfer coefficient is a crucial step in the design, operation and scale-
up of bioreactors. 
 It is possible to provide required heat transfer capacity of large scale
fermenter by methods independent of process scale up (e.g. using
refrigerator rather than cooling water, or using external heat
exchanger)
 Similarly, proper pH control can be achieved independently by
automatic addition of concentrate acids and bases in fermentation
systems with an adequate dispersing mechanism.
 Criteria for scale up operations
• Process characteristics that are suggested to remain constant
during scale up operations are:
1. Reactor geometry
2. Volumetric oxygen transfer coefficient (kL α: kL – Liquid phase
mass transfer coefficient; α – Specific gas-liquid interfacial area)
3. Maximum shear
4. Power input per unit volume of liquid (Pi/V)
5. Volumetric gas flow rate per unit volume of liquid (Q/V)
6. Superficial gas velocity (vs)
7. Mixing time
8. Impeller Reynold’s number

 Criteria 1-6 are commonly considered for scale up of bioreactors

 Criteria for scale up operations
1. Criteria1 : Reactor geometry: It is based on the fact that almost all
existent empirical or semi-empirical correlations for scale up are
developed experimentally with geometrically similar reactors of
similar scales. For non-geometric scale up the ratio of impeller
diameter to bioreactor diameter must be considered.
2. Criteria 2: Volumetric oxygen transfer coefficient: Ultimate
performance of most aerobic fermentation is often oxygen limited.
To ensure equal transfer rates at various scales of operation, scale up
is conducted with constant kL α.
3. Criteria 3: Maximum shear: This is criteria is very important with
respect to shear-sensitive microorganisms.
4. Criteria 4: Power input per unit volume of liquid (Pi/V): Equal power
per unit liquid volume has been used in many antibiotic fermentations
as the primary scale up parameter. Typical Pi/V used are 1-2 kW/m3.
Pi/V is the most successful rule for shear-sensitive microbes
5. Criteria 5 & 6: They address the importance of aeration rate for scale
up of aerobic fermentation.
 Criteria for scale up operations
Superficial gas velocity affects the mixing energy required to disperse the
gas stream. Volumetric gas flow rate per unit liquid volume has been
accepted as traditional scale up criterion even for mechanically agitated
bioreactors.