Aeration and Agitation in fermentation

Aeration and Agitation

The majority of fermentation processes are aerobic and therefore require
the provision of oxygen. The oxidation of glucose may be represented as;
C6H12O6 + 6O2 = 6H2O + 6CO2

It is not possible to provide a microbial culture with all the oxygen it will
need for the complete oxidation of glucose in one addition. Therefore, a
microbial culture must be supplied with oxygen during growth at a rate
sufficient to satisfy the organisms demand.

The oxygen demand of an industrial fermentation process is normally

satisfied by aerating and agitating the fermentation broth.
However, the productivity of many fermentations is limited by oxygen
availability and therefore, it is important to consider the factors which
affect the fermenter’s efficiency in supplying microbial cells with
oxygen.
 Aeration and Agitation

Important factors in a fermenters

Provision for adequate mixing of its contents,

Mixing in fermentation
Ø to disperse the air bubbles
 to suspend the cells
 to enhance heat and mass transfer in the medium
Ø All relate to Gas-liquid mass transfer
 Aeration and Agitation

Aeration refers to the process of introducing air to increase oxygen

concentration in liquids

Aeration may be performed by

• bubbling air through the liquid,
• spraying the liquid into the air or
• agitation of the liquid to increase surface absorption

Agitation – uniform suspension of microbial cells in homogeneous

nutrient medium
Structural components involved in aeration and agitation
• Agitator (impeller)
• Baffles
• Aeration system (sparger)
 View looking down into a stainless steel fermenter
  Agitator (impeller)
Achieve mixing objectives
• bulk fluid and gas-phase mixing,
• air dispersion,
• oxygen transfer,
• heat transfer,
• suspension of solid particles and
• maintaining uniform environment throughout vessel contents
 Baffles
Four baffles incorporated into agitated vessels of all sizes
to prevent vortex and to improve aeration efficiency
Metal strips roughly one- tenth of
vessel diameter and attached
radially to the wall
Minimizes microbial growth on
baffles and fermenter walls.
 Aeration system (sparger)
Introduces air into liquid of fermenter
Three basic types – porous sparger
1. Orifice sparger – a perforated pipe
2. Nozzle sparger – an open or partially closed pipe
3. Combined sparger-agitator
OXYGEN REQUIREMENTS OF INDUSTRIAL
FERMENTATIONS
A culture's demand for oxygen is very much dependent on the source of
carbon in the medium. Thus, the more reduced the carbon source the
greater will be the oxygen demand. From Darling~ ton's and Johnson's
equations(table 9.1), it may be seen that the production of 100 grams of
biomass from hydrocarbon requires approximately three times the
amount of oxygen to produce the same amount of biomass from
carbohydrate. However, it must be remembered that the high carbon
content of hydrocarbon substrates means that high yield factors (g
biomass g -1 substrate consumed) are obtained and the decision to use
such substrates is based on the balance between the advantage of high
 However, it is inadequate to base the provision of oxygen for a
fermentation simply on an estimation of overall demand, because the
metabolism of the culture is affected by the concentration of dissolved
oxygen in the broth. The effect of dissolved oxygen concentration on
the specific oxygen uptake rate (Qo2' mmoles of oxygen consumed
per gram dry weight of cells per hour) has been shown to be of the
Thus, maximum biomass production may be achieved by satisfying the
organism's maximum specific oxygen demand by maintaining the
dissolved oxygen concentration greater than the critical level. If the
dissolved oxygen concentration were to fall below the critical level then
the cells may be metabolically disturbed. However, it must be
remembered that it is frequently the objective of the fermentation
technologist to produce a product of the micro-organism rather than the
organism itself and that metabolic disturbance of the cell by oxygen
starvation may be advantages to the formation of certain products.
Equally, provision of a dissolved oxygen concentration far greater than the critical level
may have no influence on biomass production, but may stimulate product formation.
Thus, the aeration conditions necessary for the optimum production of a product may
be different from those favouring biomass production.
Hirose and Shibai's (1980) investigations of amino acid biosynthesis by
Brevibacterium flavum provide an excellent example of the effects of
the dissolved oxygen concentration on the production of a range of
closely related metabolites.
These workers demonstrated the critical dissolved oxygen concentration
for B. flavum to 0.01 mg dm3 and considered the extent of oxygen
supply to the culture in terms of the degree of 'oxygen satisfaction’, that
is the respiratory rate of the culture expressed as a fraction of the
maximum respiratory rate. Thus, a value of oxygen satisfaction below
unity implied that the dissolved oxygen concentration was below critical
level. The effect of the degree of oxygen satisfaction on the production
of a range of amino acids is shown in Fig. 9.2.
From Fig. 9.2 it may be seen that the production of members of the
glutamate and aspartate families of amino acids was affected
detrimentally by levels of oxygen satisfaction below 1.0, whereas
optimum production of phenylalanine, valine and leucine occurred
at oxygen satisfaction levels of 0.55, 0.60 and 0.85, respectively.
OXYGEN SUPPLY
Oxygen is normally supplied to microbial cultures in the form of air,
this being the cheapest available source of the gas. The method for
provision of a culture with a supply of air varies with the scale of the
process:
i. Laboratory-scale cultures may be aerated by means of the shake-
flask technique where the culture (50 to 100 cm3) is grown in a
conical flask (250 to 500 cm3) shaken on a platform contained in a
controlled environment chamber.
ii. ii. Pilot and industrial-scale fermentations are normally carried
out in stirred, aerated vessels, termed fermenters.
Bartholomew et at. (1950) represented the transfer of oxygen from air to
the cell, during a fermentation, as occurring in a number of steps: i. The
transfer of oxygen from an air bubble into solution. ii. The transfer of
the dissolved oxygen through the fermentation medium to the microbial
cell. iii. The uptake of the dissolved oxygen by the cell.
The rate of oxygen transfer from air bubble to the liquid phase may
be described by the equation:

Where CL : is the concentration of dissolved oxygen in the fermentation

broth(mmoles dm3 ),
t : is time (hours)
dCL/dt : the change in oxygen concentration over a time period, i.e. the
oxygen-transfer rate (mmoles O2 dm-3 h-1),
KL : the mass transfer coefficient (cm h-1)
a : the gas/liquid interface area per liquid volume (cm2 cm-3),
C* is the saturated dissolved oxygen concentration (mmoles dm-3 )
KL may be considered as the sum of the reciprocals of the resistances to
the transfer of oxygen from gas to liquid and (C* - CL) may be
considered as the 'driving force' across the resistances. It is extremely
difficult to measure both KL and 'a' in a fermentation and, therefore, the
two terms are generally combined in the term KLa, the volumetric mass-
transfer coefficient, the units of which are reciprocal time (h-1).
The larger the KLa, the higher the aeration capacity of the system. The
KLa value will depend upon the design and operating conditions of the
fermenter and will be affected by such variables as aeration rate,
agitation rate and impeller design. These variables affect 'KL' by
reducing the resistances to transfer and affect 'a' by changing the number,
size and residence time of air bubbles.
It is convenient to use KLa as a yardstick of fermenter performance
because, unlike the oxygen- transfer rate, it is unaffected by dissolved
oxygen concentration.

The dissolved oxygen concentration reflects the balance between the

supply of dissolved oxygen by the fermenter and the oxygen demand of
the organism. If the KLa of the fermenter is such that the oxygen
demand of the organism cannot be met, the dissolved oxygen
concentration will decrease below the critical level (Ccrit). If the KLa is
such that the oxygen demand of the organism can be easily met the
dissolved oxygen concentration will be greater than Ccrit and may be as
high as 70 to 80% of the saturation level.
Thus, the KLa of the fermenter must be such that the optimum oxygen
concentration for product formation can be maintained in solution
throughout the fermentation.
Thanks