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FACULTY OF ENGINEERING TECHNOLOGY

DEPARTMENT OF CHEMICAL ENGINEERING TECHNOLOGY

FERMENTATION TECH. LABORATORY


LABORATORY INSTRUCTION SHEETS

COURSE CODE BNN 30304

EXPERIMENT CODE

EXPERIMENT TITLE KLa Measurement

DATE

STUDENT NAME & MATRIK NO.

GROUP

GROUP MEMBERS

LECTURER/ INSTRUCTOR/
TUTOR

DATE OF REPORT SUBMISSION

MARKS: ATTENDANCE/DICIPLINE: /5%

RESULTS: /25%

DATA ANALYSIS: /25%

DISCUSSION & CONCLUSION: /35%

REFERENCE: /10%

TOTAL: /100%

EXAMINER COMMENTS: RECEIVED DATE AND STAMP


KOD ETIKA PELAJAR
(KEP)
JABATAN TEKNOLOGI KEJURUTERAAN KIMIA

FAKULTI TEKNOLOGI KEJURUTERAAN


Saya dengan ini mengaku bahawa saya telah menyediakan laporan ini dengan daya usaha
saya sendiri. Saya juga mengaku tidak menerima atau memberi sebarang bantuan dalam
menyediakan laporan ini dan membuat ikrar ini dengan kepercayaan bahawa apa-apa yang
tersebut di dalamnya adalah benar.

Ketua Nama:
Kumpulan
No. Matriks: (Tandatangan)

Ahli 1 Nama:

No. Matriks: (Tandatangan)

Ahli 2 Nama:

No. Matriks: (Tandatangan)

Ahli 3 Nama:

No. Matriks: (Tandatangan)

Ahli 4 Nama:

No. Matriks: (Tandatangan)

___________________________

Tandatangan Pelajar

Nama : _______________________________

No. Matrik :____________________________

Tarikh :________________________________
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

1.0 OBJECTIVE
The objectives of this experiment are :
1. To determine the KLa of a fermentation system by dynamic gassing out techniques
which depends upon the monitoring of the increase in dissolve oxygen in agitation
and aeration range.
2. To study the effect of medium viscosity on KLa value.

2.0 LEARNING OUTCOMES


1. The students will be able to understand the dynamic gassing out techniques during
fermentation process
2. The students will be able to determine KLa value of a fermentation system
3. The students will be able to investigate the effect of air flowrate, agitation speed and
salt on KLa value

3.0 INTRODUCTION / THEORY

KLa is considered as a single parameter which known as volumetric or overall 02 transfer


rate. Kca represents the oxygen transfer rate per unit volume. Whilst 02 transfer rate is
dependent on KL, a & concentration driving force.

Oxygen is an important substrate in aerobic fermentations. Since oxygen is sparingly


soluble in water, it may be the growth-limiting substrate in these fermentations. For
bacteria and yeast cultures, the critical oxygen concentration is about 10% to 50% of the
saturated DO (dissolved oxygen concentration).

Determination of KLa in a fermenter is important to establish its aeration efficiency and


quantify effects of operating variables on oxygen supply. It is also used to compare
fermenters before scale up or scale down.

Method of KLa determination

KLa can be determined using either chemical method or dynamic gassing out method.
In chemical method using sulphite oxidation technique, 0 2 transfer rate is controlled by
physical adsorption by sampling & titration of unreacted sulphite. This is inaccurate in
the presence of even low level of surface active contaminants.

In the dynamic gassing out method, the increase DO concentration of solution is


monitored during aeråtion & agitation. It is necessary to decrease the 0 2 first to the low
value. Two methods have been developed which are non-fermentative (Static Method)
and Fermentative (Dynamic Method).

This experiment applies the dynamic gassing out with static method. The oxygen
concentration of the solution is firstly lowered by gassing the liquid out with nitrogen
gas, so that the solution is "scrubbed" free of oxygen. Aeration is then initiated at a
constant flow rate and the increase in dissolved oxygen tension (DOT) is monitored
using dissolved oxygen electrode. The profile of DOT during deaeration and aeration is
shown in Figure 1. Increase in DOT during aeration can be expressed by Eq. 1;
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

3.1 APPARATUS & MATERIALS

Bioreactor including pO2 probe

Stopwatch

Calibration of dissolved oxygen electrode

Before calibration, the p02 must be polarized. The polarization must be repeated any
time the electrode is disconnected from the amplifier for more than 10 min, but may
require less time then. The calibration of p02-electrode includes zero and slope
calibration. The "zero" is the electrode's current when no oxygen is present in the
culture medium meanwhile the "slope" is usually the p02 after saturation of the medium
with air at the maximum air supply intended for the process.

The calibration of the p02 electrode involved several steps;

1 . Temperature in the culture vessel is adjusted at the operating temperature.

2. For "zero" calibration, the p02 of the culture medium can be measured before starting
the air supply. During sterilization, the medium will be degassed almost completely due
to the heat impact and thus should not contain dissolved oxygen. Alternatively, we can
supply an oxygen-free gas (such as nitrogen of 99.98 purity) to the culture medium to
displace the diSsolved oxygen until a constant p0 2 near "01' can be read at the
measurement and control system.

3. For slope adjustment, the air supply is activated and the stirring speed is adjusted to
the operating value. The medium should be optimally gassed (max. flow rate intended
for the process) and mixed. At a stable display of the measured value we can calibrate
this as "100 p02".

4. After calibration, the gas supply rate required for the start up of the intended
fermentation process can be adjusted on the rotameter of the control unit. Note that the
rotameter is calibrated according to standard conditions (temperature 20°C, with air.at
2 barabs). If it is important to maintain precise operating air flow-rates for further
calculations, it is necessary to recalculate the indicated flowrate with a correction
factor.

The calibration of the p02 -electrode is made in the culture vessel after autoclaving and
under the conditions of fermentation.
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

3.2 PROCEDURE

1. Set the agitation speed of 500 rpm and 1.0 L/min. Purge the nitrogen gas until DO.
Determine KLa of stirred tank reactor at different air flow rates (0.5, 1.0, 1.5, 2.0 and 2.5
L/min). For this experiment, set the agitation speed at 500 rpm.

2. Determine the effect of increasing agitation speed (200, 400, 600, 800 and 1000 rpm) on KL
of a 2 L stirred tank fermenter. For this experiment, set the air flow rate at 1 L/min.

3. In experiment 1 and 2, the fermenter is filled with 1.5 L of distilled water.

4. Investigate the effect of salt (NaCl) and antifoam addition to distilled water on K La. In this
experiment, add 1.5 g of NaCl to 1.5 L distilled water in a fermenter. Determine the KL at a 500
rpm and air flow rate of 1 L/min. Then, add 5 mL of antifoam in a salt solution and determine
KLa at the same agitation speed and air flow rate.
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

4.0 RESULTS & CALCULATIONS / ANALYSIS

time CL (% CL Ln(CE- time Cl (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

time CL (% CL Ln(CE- time CL (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

time CL (% CL Ln(CE- time CL (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

time CL (% CL Ln(CE- time CL (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

time CL (% CL Ln(CE- time CL (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

time CL (% CL Ln(CE- time CL (% CL Ln(CE-


(s) saturation) dCt/dt (average) CL) (s) saturation) dCt/dt (average) CL)
0 0
20 20
40 40
60 60
80 80
100 100
120 120
130 130
140 140
150 150
160 160
170 170
180 180
190 190
200 200
210 210
220 220
230 230
240 240
250 250
260 260
270 270
280 280
290 290
300 300
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

Effect of airflow rate on KLa (500 rpm)

Airflow rate (L/min) KLa(h-1)

Effect of agitation speed on KLa (air flow rate = 1 L/min)

Agitation Speed (rpm) KLa(h-1)

Effect of salt and antifoam addition on KLa


(Agitation = 500 rpm; Airflow rate = 1 L/min)

Substance KLa(h-1)

Salt

Antifoam
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

4.0 RESULTS & CALCULATIONS / ANALYSIS

1. Discuss the effect of agitation speed and airflow rate on KLa

2. Discuss on the addition of salt and antifoam on K La

5.0 DISCUSSION AND CONCLUSION

Please provide discussion and conclusion for your findings.


FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:
FACULTY: ENNGINEERING EDITION:
TECHNOLOGY 1

LABORATORY: FERMENTATION
REVISION NO:
TECH LABORATORY

EFFECTIVE DATE: 1/4/2018


EXPERIMENT: KLa Measurement
AMENDMENT DATE:

Prepared by / Disediakan oleh Approved by / Disahkan oleh :

Signature/Tandatangan:
Signature / Tandatangan :
Name/Nama: Dr Noor Akhmazillah bt Mohd
Name / Nama : Prof. Madya Dr. Ishak Baba
Fauzi
Date / Tarikh :
Date/Tarikh :