Introduction

Dissolved oxygen (DO) is an important substrate in aerobic fermentations and

may be a limiting substrate, since oxygen gas is sparingly soluble in water. At high
cell concentrations, the rate of oxygen consumption may exceed the rate of oxygen
supply, leading to oxygen limitations (Liu, 2017). The determination of KLa of a
fermenter is essential in order to establish its aeration efficiency and to quantify the
effects of operating variables on the provision of oxygen. To monitor the increase in
dissolved oxygen over an adequate range, it is necessary to first decrease the
oxygen level to a low value. Two methods can be employed to achieve this lowering
of the dissolved oxygen concentration; non- fermentative and fermentative. This
experiment uses non-fermentative method or known as dynamic gassing-out. In this
technique, the O2 concentration of the solution is lowered by gassing the liquid out
with the nitrogen gas, so that the solution is “scrubbed” free of O 2. Aeration is then
initiated at a constant air flow rate and the increase in dissolved O 2 tension (DOT) is
monitored using dissolved O2 electrode (Stanbury, et al., 2003). During batch
fermentation, KLa can be determined using non-fermentative method. To determine
the KLa, shut off air supply valve and set the agitation speed at 50. Follow the
decrease in DO with time and when the DO drop about 0% saturation, open the
valve for air supply and increase agitation speed to the set level. This also can be
done by using fixed air flow and different speed of agitation to determine the
relationship between O2 diffusion and agitation speed. From the data of C L against
time, KLa can be calculated according to the method as described below. Plot of ln
(C*- CL) versus t will produce a straight line where the slope is equal to K La. The rate
of oxygen transfer from the bubble air to the liquid phase may be described by the
equation:

dCL /dt = KLa(C* - CL )

where CL is the concentration of dissolved O 2 in the fermentation broth, in mmoles

dm-3; t is time in hours; dCL /dt is change in O2 concentration over a time period, i.e.
O2 transfer rate, in mmoles O2 dm-3h-1; KL is the mass transfer coefficient (cm h-1); a is
the gas/liquid interface area per liquid volume (cm 2cm-3); KLa is the volumetric
transfer coefficient, in reciprocal time, h -1; C* is the saturated dissolved O2
concentration, in mmoles/dm-3.

Objectives

1. To determine KLa of a fermentation system by dynamic gassing out

techniques depends upon the monitoring of the increase in dissolve oxygen in
agitation and aeration range.
2. To monitor the increase in dissolved oxygen over an adequate range, it is
necessary to fast decrease the O2 level to a low value. Two methods can be
employed to achieve this lowering of the dissolved oxygen concentration; non-
fermentative and fermentative.
3. To study the effect of medium viscosity on KLa value.
Methodology
The agitation speed was set to 500 rpm and
1.0L/min

Nitrogen gas was urged until reach 0% DO

KL a of stirred tank reactor at different air

flow rate was determined at 1.0, 2.0, 3.0,
4.0 and 5.0 L/min

Agitation speed was set at 500 rpm.

The effect of increasing agitation speed was

determined at 200, 400, 600, 800 and 1000
rpm on KL of a 2 L stirred tank fermenter

For this experiment, air flow rate was set at

1 L/min.

In experiment 1 and 2, the fermenter was

filled with 1.5 L of distilled water.
References

Liu, S. (2017). “Bioprocess engineering.” ENGINEERING BIOPROCESS Kinetics,

Sustainability, and Reactor Design (Vol. 2). https://doi.org/10.1016/s0009-
2509(02)00006-4

Stanbury, Peter F.;Whitaker, Allan;Hall, Stephen, J. (2003). Priniciple of Fermentation

Technology. Vasa, 367. https://doi.org/10.1017/CBO9781107415324.004