Measurement of kLa

 Static method (Oxygen-Balance Method)

 Sulfite Oxidation Method
 Dynamic Method
Static method (Oxygen-Balance Method):

 This technique is based on the equation for gas-liquid

mass transfer .
N A  k L  a  (C * C L )
 In the experiment, the oxygen content of gas streams
flowing to and from the fermenter are measured.
 From a mass balance at steady state, the difference in
oxygen flow between inlet and outlet must be equal to
the rate of oxygen transfer from gas to liquid:

 NA is the total quantity of gas absorbed by the system

per unit time and per unit volume
 where VL is the volume of liquid in the fermenter,
 Fg is the volumetric gas flow rate,
 CAG is the gas-phase concentration of oxygen, and
subscripts i and o refer to inlet and outlet gas
streams, respectively.
 Since gas concentrations are generally measured as
partial pressures,

 Advantage of this method is that the method can

be applied to fermenters during normal operation.
 It depends on accurate measurement of gas
composition, flow rate, pressure and temperature
 Sulfite Oxidation Method
 It is referred as Chemical method(Cooper et al., 1944)
 It is based on the oxidation of sodium sulfite to sodium sulfate
in the presence of catalyst (Cu++ or Co++).

 This reaction has following characteristics to be qualified for

the measurement of the oxygen-transfer rate:
 1. The rate of this reaction is independent of the concentration
of sodium sulfite within the range of 0.04 to 1 N.
 2. The rate of reaction is much faster than the oxygen transfer
rate; therefore, the rate of oxidation is controlled by the rate of
mass transfer alone.
 Procedure: Fill the fermenter with a 1 N sodium sulfite
solution containing at least 0.003 M Cu++ ion.
 Turn on the air and start a timer when the first bubbles
of air emerge from the sparger.
 Allow the oxidation to continue for 4 to 20 minutes,
after which, stop the air stream, agitator, and timer at the
same instant, and take a sample.
 Mix each sample with an excess of freshly pipetted
standard iodine reagent.
 Titrate with standard sodium thiosulfate solution
(Na2S2O3) to a starch indicator end point.
 Once the oxygen uptake is measured, the kLa may be
calculated by using
rA= kL a (C*AL-CAL)
where CAL is zero and C*AL is the oxygen equilibrium
concentration.
 Dynamic Method
 It is based on an unsteady-state mass balance for
oxygen.
 The main advantage of the dynamic method over
the steady-state technique is the low cost of the
equipment needed.
 During the re-oxygenation step, the system is not
at steady state.
 The rate of change in dissolved-oxygen
concentration during this period is equal to the
rate of oxygen transfer from gas to liquid, minus
the rate of oxygen uptake by the cells:

 where qox is the rate of oxygen consumption.

 We can determine an expression for qox by
considering the final steady dissolved-oxygen
concentration, CAL.
 When CAt = CAL, - dCAL/dt= 0 because there is no
change in CAL with time.
Substituting this result gives
Problem#1
 A 20L stirred fermenter containing a Bacillus thuringiensis
culture at 30°C is used for production of microbial
insecticide, kLa is determined using the dynamic method.
Air flow is shut off for a few minutes and the dissolved-
oxygen level drops; the air supply is then re-connected.
When steady state is established, the dissolved-oxygen
concentration is 78% air saturation. The following results
are obtained.
 Problem# 2
 Factors Affecting Cellular
Oxygen Demand
 Bubble Size
 Gas hold-up
 Gas Velocity
 Type of gas Sparger
 Type of agitation
 Power input to agitator
 Temperature
 Pressure
 Antifoaming agents
 Presence of cells
 Surface-active solutes.