BIOREACTORS

Enzymes
Bioconversion
products
Microorganisms
Biotransformation
Plant cells products

Recombinant
Animal cells
products

http://cotterbrothers.com/products/bioreactors-fermentation/

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.
Consumer products made by Industrial Biotechnology

https://www.bio.org/sites/default/files/files/ConsumerProducts060409.pdf
What are the advantages of Biotechnology over
Chemical synthesis?
• Mild reaction conditions-More energy efficient and cost-effective

• High regio- and stereo-selectivity

• Sustainable process-uses renewable resources as raw material

e.g. cellulose, starch, sucrose, peanut meal etc.

• Better product yields: rDNA technology for yield improvement

• No toxic and undesirable waste byproducts

• More environment friendly- Lesser CO2 emissions

 A typical bioprocess

A substrate of low value is utilized by living cells or enzymes to

generate a product of higher value.
https://slideplayer.com/slide/6437239/
 Questions to be answered for running a
Bioprocess efficiently

1. How fast will the process take place? (Reaction kinetics)

2. What changes can be expected to occur? (metabolic state, cell

physiology)

3. How can the system be operated and controlled for the maximum
yield and productivity? (Reactor design, instrumentation and
control)

4. How can the products be separated with maximum purity and

minimum cost? (Separation processes)
 Efficient bioprocess
It depends on:

• Production of organism
• Optimum conditions for desired product
formation
• Product value
• Scale of production

Includes what size of reactor, type of reactor or

method of operation are best for a given duty or
conversion?
 Points to be considered in bioreactor
designing

• control and positively influence the biological reaction

• Prevent contamination

• Capital investment and operating cost

• During fermentation: maintenance of monoseptic

conditions, optimal mixing with low & uniform shear
rates
• Aeration requirement: done by one or combination of following methods
like
– surface aeration
– direct sparging
– indirect and/or membrane aeration
– medium perfusion
– increasing the partial pressure of oxygen
– increasing the atmospheric pressure
• Maintenance of
– adequate mass transfer(oxygen)
– heat transfer
– clearly defined flow conditions
– appropriate feeding of substrates avoiding under and overdosing
– suspension of solids
– sufficient substrates, salts for nutrition, vitamins, etc.
– water and oxygen (aerobic processes)
• Gas evolution
• Product and by-product removal
 Learning Outcomes

Students will be able to explain:

• a typical bioreactor
• modes of bioreactor operation
• types of bioreactors
 What is a Bioreactor?
• Provides a controllable environment enabling the
biochemical and biomechanical requirements to
create a desirable biological product

• Enables close monitoring and control of the

reaction parameters like internal and external
mass transfer, heat transfer, fluid velocity, shear
stress etc.
 Basic features of a bioreactor
• Head space volume
• Agitator system
• Oxygen delivery system
• Foam control system
• Temperature & pH control system
• Sampling ports
• Cleaning & sterilization system
• Charging and empyting lines
 A typical bioreactor

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http://bioreactoritumenarik.blogspot.in/2012/12/in-first-day-meet-mr.html
http://www.yourarticlelibrary.com/micro-biology/bioreactors-fermenters-function-designs-and-types/33628/
 Typical Bioreactor Parts
Agitator -It achieves the following objectives:
❑ Bulk-fluid and gas-phase mixing
❑ Air dispersion
❑ Oxygen transfer
❑ Heat transfer
❑ Suspension of solid particles
❑ Maintenance of uniform environment throughout the vessel

The agitator can be top or bottom driven which could be basically magnetic /
mechanically driven.

There are two types of impellers, depending on the flow regime created
Axial flow impeller and Radial flow impeller

"http://www.liquid-mixing.com/impeller.html" , https://cercell.com/support/bactovessel-details/turbine-principles/,
https://www.postmixing.com/mixing%20forum/impellers/pr.htm.
 Baffle – Metal strips roughly 1/10th of the vessel diameter, attached
radially to the fermenter wall (4-8 in no.).

❑ Prevents vortex formation

❑ Improves aeration efficiency
❑ Extra cooling coils may be attached to baffles to improve
cooling.
❑ Scouring action around and behind the baffles minimises
microbial growth on the fermenter wall.

Sparger – Introduces air into the fermenter broth.

❑ Provides oxygen to the growing cells

❑ Promotes mixing of the reactor contents
❑ Reduces power consumed to achieve desired homogeneity

https://www.postmixing.com/mixing%20forum/Macro/Fluid%20Motion/process_intensifier/standards.htm?hcb=1
 Cooling/Heating Jacket –

❑ Annular area for circulation of constant temperature water.

❑ Circulating water is maintained in a separate Chilled Water Circulator.
❑ Contact area of jacket provides adequate heat transfer area.

https://www.glaskeller.ch/en/products/reaction/reaction-unit-pilot-plant-fully-functional-stand/,
Basic control systems for the operation of the bioreactor :

Temperature measurement and control –

❑The measurement of the temperature of the bioreactor is done by a
thermocouple
❑The measured signal sent to the controller is used to calculate the error
❑Heating or cooling finger of the bioreactor is activated to decrease the error
❑The measured temperature is brought close to the set point

pH measurement and control –

❑The measurement of pH in the bioreactor is done by the autoclavable pH probe.

❑The measured signal is used to calculate the error.
❑ Activation of the acid or alkali pump to reduce the error.
❑Calibration with buffers in the pH range of the experiment.

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Dissolved oxygen controller –
❑ The dissolved oxygen in the bioreactor broth is measured by a dissolved oxygen
probe
❑Calibrated for its zero and hundred percent values.
❑ DO control: Increasing air-flow rate or impeller speed or purging pure oxygen.

http://www.usbmeter.com/Theory_DO.htm
Foam –formed due to entrapment of air in micelles made of proteinaceous material from
the medium or from cell lysis which have detergent like properties.

Disadvantages:
❑It can lead to blockage of exit filters, contamination,
❑pressure rise in the reactor, medium loss.

Control- Foam sensor

Change in air space-capacitance due to the foam changes the
magnitude of the current.
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❑Act by dewetting/streching.
❑Act on Gas-liquid interface.
❑High spreading efficiency on foam surface-destabilize the foam
❑Reduces kLa inside the reactor
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.

In terms of process requirements, they are of the following

types:
(i) aerobic, (ii) anaerobic, (iii) solid state (iv) immobilized
cell bioreactors.
Classification of bioreactors
Based on the presence or absence of oxygen and requirement of
stirring
Anaerobic fermentation
❑ These reactors do not require aeration
❑ Initial preparation of inoculum may requires aeration.
❑ The gas released during fermentation is sufficient to provide mixing.
❑ The product recovery may also require anaerobic conditions

Non-stirred non-aerated bioreactors

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Aerobic fermentations
These reactors should have adequate provisions for supply of sterile air and also need a
mechanism of stirring up and mixing the medium and cells.

• Non-stirred aerated reactors (Airlift, bubble column reactors)

• Stirred and aerated reactors (stirred tank reactors)

Martínez, A. M. M., & Silva, E. M. E. (2013). Airlift bioreactors: hydrodynamics and rheology application to secondary metabolites
production. Mass Transfer: Advances in Sustainable Energy and Environment Oriented Numerical Modeling, 387,
https://www.slideshare.net/angelsalaman/bioreactor-basis-34000053
Air lift bioreactors

❑The stirred tank bioreactors lack well defined flow of air.

❑Air is pumped from below.
❑Bubbles rise up through the draught tube and drags the surrounding fluid.
❑ Air stir up the contents.
Advantages
❑ Less friction
❑ Less cost and energy requirements
❑ Easy scale up.
Disadvantages
❑ Difficulty of sterilization
❑ Efficiency of mixing is low
 Modes of bioreactor operation
• Batch reactors
• Fed batch
• Continuous

❑ Initial Capital Expenditure

❑ Easy operation
❑ Less downtime
❑ Batch to batch variations in product quality
❑ Productivity and yield
❑ Maintenance of reactor components (probes)-sterilization frequency
❑ Requirement of skilled personnel
❑ Instrument for Feedback control-cost addition
❑ Quality of the culture (Mutations and clonal selection)
Díaz-Montaño, D. M. (2013). Continuous agave juice fermentation for producing bioethanol
 Based on the method of cultivating the culture

• Solid-state (cultivation in the absence of free water; pretreatment of substrate;

difficult to control process parameters)

Applications:
Antibiotics, Food additives,
Biocontrol agents,
Food processing
Biopulping/bioremediation,

Disadvantages
❑Channelling and flooding
❑High pressure drop
❑Oxygen transfer limitation
❑Temperature and pH control is difficult
❑non-uniform environment (nutrient concentration gradients)
❑Shear damage (movement of large substrate particles)
http://slideplayer.com/slide/9920824/
Immobilized cell bioreactors
These are based on immobilized cells.

Advantages

❑ Re-utilization of cells
❑ Can operate at high cell densities
❑ Reduction in contact time leading to increased
volumetric productivity.
❑ Simplified downstream processing.
❑ A continuous process increases volumetric
productivity
❑ Allows removal of metabolic inhibitors.
❑ Reduces problems like aggregation, shear
damage and foaming.

http://english.qibebt.cas.cn/rh/rp/200909/t20090908_36482.html,
https://scialert.net/fulltext/?doi=pjbs.2011.526.532
Fluidized bed reactors

Advantages
❑ Heat and mass transfer is efficient
❑ The mixing between the liquid, solid and gaseous phases is effective.
❑ Low shear rates: suitable for plant cells and mammalian cells.

https://slideplayer.com/slide/7123360/
 Hollow fiber membrane bioreactors
Fibers: cellulose acetate, acrylic polymers, etc.

Advantages
❑ Extracellular products is separated from cells.
❑ Productivity is high.
❑ Easy Scale up: several parallel fiber units.

Disadvantages
❑ Sometimes, the pores get plugged.
❑ Cell growth around the lumen can sometimes
distort and rupture the fibers.
❑ If the toxic products happen to accumulate
in the fiber it may inhibit the growth of microbes.
https://cellculturedish.com/hollow-fiber-provides-a-sweet-spot-for-several-
biomanufacturing-applications/
https://www.fibercellsystems.com/advantage/, https://americanlaboratory.com/914-
Application-Notes/114793-The-Hollow-Fiber-Bioreactor-and-Cell-Co-Cultivation
https://www.slideshare.net/sarahairasantos/lecture-9b-scaling-up
 Disposable bioreactors
The Wave bioreactor
Principle of a wave and undertow mechanism provides mixing and
aeration to the culture.

Ducos, J. P., Terrier, B., Courtois, D., & Pétiard, V. (2008). Improvement
of plastic-based disposable bioreactors for plant science needs.
Phytochemistry Reviews, 7(3), 607-613.
 Bioreactor design and analysis
• What size of reactor, type of reactor or method of
operation are best for a given duty or conversion?

• Performance measures:
– Reaction time is the measure of the processing rate in a batch
reactor

– Space time (: time required to process one reactor volume of

feed) and space velocity (no. of reactor volume treated in unit
time) are measures for performance of flow reactors