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EXPERIMENT 6

EFFECT OF AGITATION ON KLa IN A BIOREACTOR


1.0 OBJECTIVES
1.1 To introduce the concept and importance of oxygen transfer for biological and
environmental systems.
1.2 To determine the effect of agitation on dissolve oxygen in kLa measurement.
1.3 To develop a dynamic approach in kLa determination.

2.0 COURSE OUTCOME


CO3: Ability to design a stirred tank bioreactor according to the specific application.

3.0 INTRODUCTION
The oxygen supply into the fermentation broth constitutes one of the decisive factors of
cultivated microorganism’s growth and can play an important role in the scale-up and
economy of aerobic biosynthesis systems. The aeration efficiency depends on oxygen
solubilization and diffusion rate into the broth, respectively and on the bioreactor capacity to
satisfy the oxygen demand of microbial population. The oxygen mass transfer can be
described and analyzed by means of mass transfer coefficient, kLa. It represents the most
important parameter implied on the design and operation of mixing-sparging equipment of
the bioreactors. The kLa values are affected by a lot of factors, such as geometrical and
operational characteristics of the vessels, media consumption, fluid type, concentration and
microorganisms’ morphology.

4.0 THEORY
4.1 Unsteady state method (Dynamic method)

Fig 1: Variation of oxygen tension for dynamic measurement of kLa


One of the several methods to determine oxygen transfer coefficient of a bioreactor is based
on a dynamic approach of oxygen measurement (Taguchi and Humphrey, 1966). This is
based on an oxygen balance performed across an aerated bioreactor in which a living
culture is actively growing. A material balance gives:

[Oxygen transferred from gas phase] – [Oxygen consumed by cells] = [Accumulation]

 k L aC AL  C AL   qO X
dC AL *
2 Eq 1
dt
where CL is dissolved oxygen concentration in the liquid phase, CL* is dissolved oxygen
concentration in equilibrium with partial pressure of oxygen in the air, kLa is the volumetric
oxygen transfer coefficient, qO is the rate of oxygen used per unit mass of organisms, and X
2

is the mass concentration of living cells.

qO2 X can be determined by considering the final steady state dissolve-oxygen


 
concentration, C AL . When CAL= C AL , dCAL/dt = 0 because there is no change in CAL with

time.

Therefore from Eq 1 :

k L aC AL
*
 C AL

  qo2 X Eq 2

Substituting this result into Eq 1 and canceling the kLaC*AL terms gives:

 k L aC AL  C AL 
dC AL 
Eq 3
dt

Assuming kLa is constant with time, integration Eq 1 between t1 and t2 will give:
 C AL

 C AL1 
ln     k L a(t 2 t1 ) Eq 4
 AL
C  C AL 2 

kLa can be estimated when is plotted against as


shown in the Figure 2, and kLa is given by the slope of the plot.

Fig 2 : Evaluating kLa using dynamic method

Other types in kLa measurement are steady state and sulfite test.

5.0 MATERIALS AND EQUIPMENTS


5.1 2 L Mammalian bioreactor
5.2 Nitrogen gas
5.3 Air

6.0 PROCEDURE
6.1 Switch on the mains power on the bioreactor unit.
6.2 Fill the bioreactor with 1 L of distilled water.
6.3 The parameters that need to be adjusted are:

Stirrer speed Temperature Aeration rate


(rpm) (OC) (l/min)
200 30 0.5
600 30 2.0
800 30 3.0

6.4 Record the dissolve oxygen pattern.


7.0 RESULT AND CALCULATION
7.1 Record all the results.

7.2 Plot the graph of against and evaluate the


slope.
7.3 For different stirrer speed, calculate the kLa value.
7.4 Plot kLa against agitation speed.

8.0 QUESTIONS
8.1 How do temperature and agitation affect the dissolved oxygen measurement and the
culture?
8.2 How to perform material balance on an actively growing yeast culture?
8.3 Does the DO affect yeast growth? If yes, explain how it is affecting the growth of yeast?
8.4 What are the correlations between kLa and viscosity of the culture?
8.5 What does the value of kLa reflects?

9.0 CONCLUSION
State your conclusion based on your experimental result.