Bioreactor System

Major Function:
to provide controlled environment for
growth of microorganism (or mixture) to
obtain desired product.
Points to be considered in designing and
constructing a bioreactor:
• Microbiological and biochemical characteristics of
cell systems.
• Hydrodynamic characteristics of the bioreactor.
• Mass and heat characteristics of the bioreactor.
• Kinetic of cell growth and product formation.
• Genetic stability characteristics of the cell system.
• Aseptic equipment design.
• Control of bioreactor environment (macro and
microenvironments).
• Implication of bioreactor design on downstream
product separation.
• Capital and operating costs of the bioreactor.
• Potential for bioreactor scale up.
Types of Bioreactor

Vary in size and complexity,

test tube (10 ml) to
computer controlled
fermenters (>100 m3).
1. Standing Cultures

-Little or no power used for aeration.

-Aeration- dependent on transfer of oxygen
through the still surface of the culture.
-Poor rate of oxygen transfer – due to small
surface area.
-Used in small-scale (oxygen supply is not
critical).
-Ex: Biochemical tests for identification of
bacteria (5-10mL of test tube)
a. T- Flasks

-Small scale culture of animal cells

horizontally to increase the surface area for
oxygen transfer.
-Surface aeration rate can be increased
using larger volume flasks.
b. Fernback Flasks
c. Surface Culture

-Not restricted to the laboratory.

-In some countries where electricity is unreliable,
citric acid is produced using surface culture
techniques.
-Aspergillus niger mycelia are grown on surface of
liquid media in large shallow trays. The medium is
neither gassed nor agitated.
 Ex: Aerobic solid substrate fermentations
(biomass is grown on solid biodegradable
substrates such as water softened bran, rice or
barley, solids continuously or periodically turned
over to improve aeration and to regulate culture
temperature).

Ex:
1. Production of koji by Aspergillus oryzae
on soya beans, (part of soya sauce
process).
2. Mushroom cultivation.
2. Shake Flask

-Small scale cell cultivation (continuous

shaking of culture fluid), higher oxygen
transfer rates.
-Shaking breaks liquid surface and provides
greater surface area for oxygen transfer.
-Increased rates of oxygen transfer are also
achieved by entrainment of oxygen bubbles
at the surface of the liquid.
 a. Factors affecting KLa

-Shaking speed
-Liquid volume
-Shake flask design
KLa decreases with
liquid volume KLa is higher when

-Baffles baffles are present

KLa increases with

liquid surface area

Unbaffled flask Baffled flask

Baffle
3. Mechanically Stirred Tank
Bioreactor
-Most important bioreactor for
industrial application
(pharmaceutical industry).
-Low capital cost, low operating
costs, best understood and
flexible.
-Non-sparged mechanically
agitated bioreactors can supply
sufficient aeration for microbial
fermentations with liquid
volumes up to 3 L.
-However, stirring speeds of up to 600 rpm may
be required before the culture is not oxygen
limited.
-Oxygen transferred from head-space.
-Agitation continually breaks the liquid surface
and increases surface area for oxygen transfer
Sparged bioreactor:
->3L, air sparging is required for effective oxygen
transfer.
-Agitation breaks up bubbles and increase KLa
-Required lower agitation speeds for aeration
efficiencies comparable to non-sparged fermenters.
-Air-sparged bioreactors can have liquid volumes of
greater than 500,000 L
4. Bubble Driven Bioreactors

-Sparging without mechanical agitation can

also be used for aeration and agitation.

-Two classes: bubble column fermenters

and airlift fermenters.

- Bubble driven bioreactors are commonly

used in the culture of shear sensitive
organisms such as moulds and plant cells.
- An airlift
fermenter
differs from
bubble column
bioreactors by
the presence of
a draft tube
provides better
mass and heat
transfer
efficiencies and
more uniform
shear
conditions.
 - The main functions of draft tube:

1.  Increase mixing through the reactor

2.  The draft tube enhances axial mixing throughout the whole
reactor
3.   Reduce bubble coalescence.

Due to circulatory effect, draft tube induces in

the reactor. Circulation occurs in one direction
and hence the bubbles also travel in one
direction.
5. Airlift Bioreactor

Three regions: air-riser, down-comer and

disengagement zone.
 6. Fixed Bed Bioreactors

Cells are immobilized by absorption on or entrapment

in solid, non-moving solid surfaces(e.g: plastic
blocks, concrete blocks, wood shavings or fibrous
material such as plastic or glass wool).
 7. Packed Bed Bioreactor
Rate of mass transfer between cells and medium
depends on flow rate and on thickness of biomass film
on or near the surface of the solid particles.
 
Problems: poor mass transfer rates and clogging.
 
Used commercially with enzymatically catalysts and
with slowly or non-growing cells.
8. Trickle Flow Bioreactors
Used in aerobic treatment of sewage. Oxygen
transfer is enhanced by ensuring cells are covered
by only a very thin layer of liquid, thus reducing
the distance over which the dissolved oxygen must
diffuse to reach the cells.
The liquid
medium
trickles
over the
surface of
the solids
on which
the cells
are
immobilized Oxygen diffuses
through a thin layer of
liquid surrounding the
cells.
9. Fluidized Bed Bioreactors
Maintaining high biomass concentrations, good
mass transfer rates in continuous cultures.

Mixing is assisted by the action of a pump

cells or enzymes are immobilized in and/or on the
surface of light particles.
 2.3 Application of
Bioreactor in Industry
Downstream engineering is another component parts
of the fermentation process that is needed for product
recovery and purification. It must be remembered that
the fermentation and product recovery are integrated
parts of an overall process. Because of interactions
between the two, neither stage should be developed
independently, which might result in problems and
unnecessary expenses. The extraction and purification
of fermentation product may be difficult and costly.
Recovery costs of microbial products may vary from
as low as 20% to as 60% of the total manufacturing
costs.
In general, the recovery process could be divided into several
stages.

•particles
The first stage of the recovery process is to remove large
and microbial cells from the culture broth. This step can
be done either by centrifugation, sedimentation or filtration.
•extracted
In the next stage, the broth is fractionated or
into major fractions using adsorption or ion-
exchange chromatography, liquid-liquid solvent
extraction or precipitation.
•containing
Afterwards, the product-
fraction is
purified by fractional
precipitation, furthermore
precise chromatographic
techniques and
crystallization.

•recovery
The chosen process for
will depend on the
specific products and the
equipment which is
available.
Stock culture of INOCULUM Powderized biomass can be used
PRODUCTION as biosorbent for removal of heavy
A. flavus DEVELOPMENT
metal from industrial effluent
Seed
Production
fermenter
fermenter
Cell or
Biomass
DOWNSTREAM

CELL
Culture SEPARATION
fluid

Cell-free
Shake flask supernatant

MEDIUM STERILIZATION PRODUCT

EXTRACTION EFFLUENT
TREATMENT
MEDIUM FORMULATION
PRODUCT
PURIFICATION
Medium raw
materials
Glucose/Sago Starch, Yeast extract,
Minerals and Methanol PRODUCT PACKAGING Kojic Acid

A generalized schematic representation of kojic acid fermentation process.

 Process Time: 142 h (6 days)

Fermentation process

Maintain in freeze -
dried form and stored
at –80oC

2. Inoculum

Medium preparation, 2 x 1000 ml flasks, sterilization,

incubation Transfering
Medium preparation,
10 h Fermentation Process
sterilization of 2 x 10 L inoculum
12 h, 300 rpm, 35oC, into
fermenter, calibration of pH pH 7 fermenters
probe 3. 10 L Seed
3h Ferme nter

Inoculation of seed
culture into pilot
scale fermenter
4. 100 L Production
fermenter

Medium preparation, sterilization of Fermentation Process Transfering the

100 L fermenter, calibration of pH 12 h, 150 rpm, 35oC, pH 7
probe
culture into
4h production fermenter

6. Centrifugation

5. 1000 L Production
Separation of cell biomass
fermenter
from culture broth, Medium preparation, sterilization of Fermentation Process
2h 100 L fermenter, calibration of pH 12 h, 150 rpm, 35oC, pH 7
probe
4h

Cell Biomass
9. WASTE
WATER
TREATMENT
To Downstream Processing Stage
7. Freeze - Powderized
drying Active Cell
8. Formulation and
Lactic Acid Purification Packaging
FINAL PRODUCT OF ACTIVE LIVE
-80oC, 102 h, cell pellet mix with stabiliser and
protective agent, Drying in freeze-dryer CELL OF MICROORGANISM READY
FOR APPLICATION
LACTIC ACID
Mix with stabiliser, e.g starch, rice
bran. Vacuum Pack, 6 h

Dowstream processing