Professional Documents
Culture Documents
Chapter 6
Introduction
Most industrial fermentation are carried out as
pure cultures-selected strains are allowed to
grow.
Foreign microorganisms exist in the medium or
any parts of the equipment, the production
organisms have to compete with the
contaminants (limited nutrients).
Foreign microorganism can produce harmful
products-limit the growth of the production
organism.
B4 starting fermentation, medium + all
fermentation equipment – free from any living
organism (completely sterilized).
The aseptic condition - maintained
Sterilization Methods
Most effectively-removal of
microorganisms from air or other gases.
In the case of liquid solutions-used with
medium or products which easily
destroyed by heat: human & animal
serums, enzymes.
Batch Sterilization
Method:*
steam sparging
electrical heating
steam heating (constant pressure condensing
steam).
• Sterilization cycles are composed of heating,
holding & cooling
• Total Del factor:
Heating & cooling-the values by the methods used
for heating & cooling
Holding- the values by the length of the controlled
holding period.
Continuous Sterilization
Advantages:
simplifies production planning, thus allowing max
plant utilization & minimum delays
provides reproducible conditions
Can be operated at a high temperature (140°C
instead of 121°C in batch sterilization)-the
sterilization time can be shortened (holding time 1-2
min)
requires less steam by recovering heat from the
sterilized medium.
require less cooling water
easier to automate the process – less labor
intensive
• Consist 3 main sections: heating, holding & cooling
Heating section:
- 2 methods:
direct steam injection
indirect heating in the shell-and –tube or plate-
and-frame heat exchanger
- Direct heating more effective-no barrier
between the medium & heat source.
- the steam injection heats the medium to the
peak sterilization temperature quickly
(sterilization during heating period is negligible)
- Indirect heating- the plate-and-frame HE
more effective than shell-and-tube type for
heat transfer due to its larger heat transfer
area.
- the plate-and-frame favorable for the
sterilization of a high viscous system
Plate-and-frame
Shell-and-tube
Holding Section:
- the heated medium passes through a holding
section, usually composed of a long tube.
- maintained in adiabatic conditions
- if the heat loss in the section is negligible, the
temperature can be assumed to be constant.
Cooling Section:
- a quench cooler with adequate heat removal
capacity is effective.
- flash cooling- inject the hot medium through an
expansion valve into the vacuum chamber
- Both of these take a very short time
- therefore, the sterilization during the cooling
period can be assumed to be negligible.
- shell-and-tube & plate-and-frame HE can also
be employed for cooling.
Air Sterilization
For aerobic fermentations, air needs to be supplied
continuously
Typical aeration rates for aerobic fermentation are 0.5-1.0
vvm (air volume per liquid vol per min).
This requires an enormous amount of air
Therefore, not only the medium but also the air must be
free from microbial contaminants.
All of the sterilization technique for medium can be
employed for air.
Sterilization of air by means of heat is economically
impractical & ineffective due to the low heat transfer
efficiency of air compared with liquids.
Most effective technique for air sterilization- fibrous or
membrane filters
The cotton plug, routinely used as a closure for tubes or
flask of sterile solution-good example of removal
microorganisms from air by a fibrous filter.
A simple air filter can be made by packing cotton
into a column.
With cotton filters the pressure drop is high and
wetting can be a breeding ground for the
contamination
Glass fibers are favorable as filter medium- give a
lower pressure drop & less liable to wetting or
combustion
Modern fibrous filter systems are cylinders made
from bonded borosilicate microfibers
With fibrous filters, airborne particles are collected
by the mechanism of:
Impaction
Interception
Diffusion
Sterilization of Fermenter
The fermenter has to be sterilized separately b4
the sterile medium is added to it.
By heating the jackets or coils of fermenter with
steam and sparging steam into the vessel
through all entries, apart from the air outlet from
which steam is allowed to exit slowly.
Steam pressure-15 psi for approximately 20 min
Sterile air is sparged into the fermenter after
cycle is complete & +ve pressure is maintained.
Sterilization of Feeds
A variety of additive may be administered to a
fermentation during the process & it is essential
that these materials are sterile
The sterilization method depends on the nature
of the additive, the volume and feed rate at
which it is administered.
Ex: if fed in large quantities then continuous
sterilization may be desirable.
Batch sterilization of feed liquids normally
involves steam injection into the material held in
storage vessels.
Sterilization of Liquid Wastes
Process organisms which have been engineered
to produce ‘foreign’ products & therefore contain
heterologous genes are subject to strict
containment regulations.
Thus, waste biomass of such organisms must be
sterilized b4 disposal.
By either batch or continuous.
Batch sterilization- sparging the steam into
holding tanks
Cont sterilization- employ the type of heat
exchangers