ABSTRACT

Biochemical processes is one of the process occurred in the living things’ cells.
Generally there are many processes in the biochemical process and of it is the aerobic
diffusion as in the bioreactor stirred tank experiment. The main objective of this experiment
is to identify the mass transfer coefficient for each parameter being investigated, such as the
effect of the agitation speed, the aeration rate as well as the temperature of the solution into
the bioreactor stirred tank. The agitation speed effect can be identified by fixing the aeration
rate being supplied into the reactor to be 2 L/min and the fixed temperature is 30˚C. The
agitation speed effect can be started at the 200 rpm of speed and the nitrogen can be supplied
into the bioreactor so that the oxygen concentration can be reduced until it is saturated. Only
after that, the flow is stopped and the oxygen can be supplied again at the fixed constant as
mention before. The value of the concentration contained in the reaction is measured by
recording the pO2 reading at the panel and this value is the C L value that will be used to plot
the graph by using (Equation 1) mentioned in the Theory section. Finally, the steps are
repeated for different agitation speed of 400 rpm, 600 rpm, 800 rpm and 1000rpm. The
overall step also can be used in determining the effect of the aeration rate but with the
different fixed variable, which is by using the agitation speed of 400 rpm and the temperature
of 30˚C. The aeration rate can be varies from 0.5 L/min, 1.0 L/min, 1.5 L/min and 2.5 L/min.
Not only that, the overall step also can be used in determining the effect of the temperature
but with the different fixed variable, which is by using the agitation speed of 400 rpm and the
aeration rate of 2 L/min. The temperature can be varies from 30˚C, 35˚C, 40˚C, 45˚C and
50˚C. From the graph of ln (C*-CL) against time plotted, the equation for each line can be
measured since the slope of the line is the kLa value for that parameter. From the results, it
shown that the higher agitation speed of the impeller gives the highest the value of the mass
transfer coefficient, kLa. Besides, the highest aeration rate being supplied into the bioreactor
tank also gives the highest value of the mass transfer coefficient, kLa. Finally, the highest
temperature of a reaction also gives the highest value mass transfer coefficient compared to
the lower temperature.
INTRODUCTION

There are many biochemical reactions existed in this life since this reaction is much
more efficient rather than another reaction. Biochemical reactions are one of the chemical
reactions that take place in living things’ cells. The reactions that happened in an organism
are called as metabolism and this is including both exothermic as well as the endothermic
reactions (Foundation, 2017).

Generally, there are few types of biochemical reactions such as oxidation and
reduction, movement of functional groups within or between molecules, addition and
removal of water and also the bond-breaking reactions (CliffNotes, 2016). Mostly, these
reactions require some amount of oxygen in order for it to operate efficiently and to produce
the product of the reaction. In order to achieve the aim, the dissolved oxygen concentration
becoming the most needed variable to be supplied into the biochemical reaction continuously.
This is because it is to ensure that the oxygen supplied is maintained throughout the
experiment while the organism is consuming the oxygen.

Physically, the oxygen is purged into the bioreactor and transferring the air bubbles
into the solution. After that, it will breaking up and mixed well in the solution by the stirrer.
For this experiment, there parameters is used to determine the difference in the mass transfer
coefficient, kLa value which is the effect of agitation speed, aeration rate and also the
temperature. The importance of studying the mass transfer coefficient shows that the
effective transfer process is by ensuring the mass transfer rate of oxygen to the liquid must
equal or exceeding the rate at which the cells take up the oxygen. This proved that the process
conditions must adequate sufficient amount of oxygen for the cell to consume freely as it will
grow (Kane, 2012).

Basically, aerobic reaction is using the following formula to utilize graphical method:

dCL/dt = Oxygen Transfer Rate (OTR) – Oxygen Uptake Rate (OUR)

dCL/dt = kLa (C* - CL) – qO2 . X

OBJECTIVES

1. To investigate the effect of agitation speed on volumetric oxygen transfer rate 𝐾𝐿 𝐴 , in

a stirred tank bioreactor.
2. To investigate the effect of aeration rate on volumetric oxygen transfer rate 𝐾𝐿 𝐴 , in a
stirred tank bioreactor.
3. To investigate the effect of the temperature on volumetric oxygen transfer rate 𝐾𝐿 𝐴 , in
a stirred tank bioreactor.

THEORY

The main objective of running this experiment is absolutely to find the mass transfer
coefficient, kLa for each parameter, which is agitation speed, aeration rate and also the
temperature. Firstly, a graph is plotted between ln (C* - CL) which shows how much is the
amount of oxygen gas contained in that certain reaction at certain period of time. The y-axis
value for the first graph is:

y-axis = ln (C* - CL) (Equation 1)

this values will be plotted against time recorded for each parameter until it reaches 100% of
pO2, or at least it reached its constant value of pO2 which showed that it has already reaching
its ‘100% value’. After the graph is drawn, the equation for each line is measured to identify
the slope for each line.

y = mx + c, where the m is = kLa (Equation 2)

From the equation, the mass transfer coefficient, kLa can be directly calculated from
the line’s slope and the value is converted into the unit of h-1. Then, these values for each line
is plotted to show the relation between the different in parameter with their respected value of
mass transfer coefficient.
APPARATUS

Figure 1: Bioreactor stirred tank.

No. Descriptions
1. Bioreactor vessel
2. Aeration meter
3. Reader for pO2
4. Calibration meter
PROCEDURE

LAB 1: The effect of agitation speed study.

1. The agitation of bioreactor is set to 200rpm.

2. Oxygen concentration of the solution is lowered by gassing the liquid out with 𝑁2 at 9
L/min until saturate and stop the flow.
3. 𝑂2 to the bioreactor is supplied at flow rate of 2.0 L/min until saturated then the flow
is stopped.
4. 𝐶𝐿 value is obtained at constant time interval during aeration
5. Steps 2 to 4 is repeated for different agitation speed (400rpm, 600rpm, 800 rpm and
1000rpm).
6. The 𝐾𝐿 𝐴 value of stirred tank reactor is calculated at different agitation speed.

LAB 2: The effect of aeration rate study.

1. The aeration of the bioreactor is set to 0.5L/min.

2. Oxygen concentration of the solution is lowered by gassing the liquid out with 𝑁2 at 9
L/min until saturate and the flow is stopped.
3. 𝑂2 to the bioreactor is supplied to the bioreactor at fixed agitation rate of 400 rpm and
at temperature of 30˚C until saturated and the flow is stopped.
4. 𝐶𝐿 value is obtained at constant time interval of 5 seconds during aeration.
5. Steps 2 to 4 is repeated for different aeration rate (1 L/min, 1.5 L/min and 2.5 L/min).
6. The 𝐾𝐿 𝐴 value of stirred tank reactor is calculated at different aeration rate.

LAB 3: The effect of temperature study.

1. The temperature of the bioreactor is set to 30˚C.

2. Oxygen concentration of the solution is lowered by gassing the liquid out with 𝑁2 at 9
L/min until saturate and the flow is stopped.
3. 𝑂2 to the bioreactor is supplied to the bioreactor at fixed agitation rate of 400 rpm and
at aeration rate at 2 L/min until saturated and the flow is stopped.
4. 𝐶𝐿 value is obtained at constant time interval of 5 seconds during aeration.
5. Steps 2 to 4 is repeated for different temperature (35˚C, 40˚C, 45˚C and 50˚C).
6. The 𝐾𝐿 𝐴 value of stirred tank reactor is calculated at different temperature.
RESULTS

A. The effect of agitation speed with constant of the temperature (30˚C) and the
aeration rate (2L/min).
Table 1: The reading measured from the experiment for agitation speed effect.

Time Agitation speed (rpm)

(s) 200 400 600 800 1000
0.00 0.00 0.00 0.00 1.70
0
1.48 3.06 4.87 5.09 2.72
5
4.32 8.03 14.70 16.00 10.70
10
9.91 13.30 23.80 29.80 25.40
15
11.90 19.70 32.00 43.70 41.40
20
15.40 26.40 41.10 53.80 56.90
25
19.30 32.60 50.40 69.80 66.20
30
23.90 38.60 57.90 72.80 77.30
35
27.90 45.80 64.80 79.30 83.50
40
31.80 50.90 70.50 84.10 88.70
45
35.80 55.80 75.70 88.40 92.00
50
39.50 60.60 80.50 91.40 94.80
55
42.70 65.20 84.00 94.00 97.00
60
46.20 69.20 86.80 95.70 98.00
65
49.90 73.00 89.40 97.00 99.00
70
53.00 76.20 91.60 98.30 99.80
75
55.80 79.20 93.50 99.10 100.00
80
58.70 81.60 95.10 99.80
85
61.40 83.90 96.40 100.00
90
63.90 85.90 97.40
95
66.50 87.60 98.30
100
 68.70 89.20 98.90
105
70.60 90.50 99.50
110
72.70 91.80 99.90
115
74.50 93.00 100.00
120
76.30 94.10
125
77.90 95.10
130
79.40 95.90
135
80.70 96.60
140
81.90 97.20
145
83.10 97.80
150
84.40 98.30
155
85.50 98.60
160
86.60 99.10
165
87.50 99.40
170
88.40 99.70
175
89.20 100.00
180
90.20
185
90.90
190
91.60
195
92.20
200
92.90
205
93.50
210
94.00
215
94.60
220
95.00
225
95.40
230
 95.80
235
96.30
240
96.70
245
97.00
250
97.30
255
97.60
260
97.90
265
98.10
270
98.40
275
98.60
280
98.70
285
98.90
290
99.10
295
99.30
300
99.40
305
99.60
310
99.80
315
99.90
320
100.00
325
Table 2: The value calculated to plot the graph of ln (C*-CL) for agitation speed (rpm)
against time.

Time ln (C* - CL) ln (C* - CL) ln (C* - CL) ln (C* - CL) ln (C* - CL)
(s) at 200 rpm at 400 rpm at 600 rpm at 800 rpm at 1000 rpm
4.61 4.61 4.61 4.61 4.59
0
5 4.59 4.57 4.56 4.55 4.58

10 4.56 4.52 4.45 4.43 4.49

15 4.50 4.46 4.33 4.25 4.31

20 4.48 4.39 4.22 4.03 4.07

25 4.44 4.30 4.08 3.83 3.76

30 4.39 4.21 3.90 3.41 3.52

35 4.33 4.12 3.74 3.30 3.12

40 4.28 3.99 3.56 3.03 2.80

45 4.22 3.89 3.38 2.77 2.42

50 4.16 3.79 3.19 2.45 2.08

55 4.10 3.67 2.97 2.15 1.65

60 4.05 3.55 2.77 1.79 1.10

65 3.99 3.43 2.58 1.46 0.69

70 3.91 3.30 2.36 1.10 0.00

75 3.85 3.17 2.13 0.53 -1.61

80 3.79 3.03 1.87 -0.11 -

85 3.72 2.91 1.59 -1.61

90 3.65 2.78 1.28 -

95 3.59 2.65 0.96

100 3.51 2.52 0.53

105 3.44 2.38 0.10

110 3.38 2.25 -0.69

115 3.31 2.10 -2.30

120 3.24 1.95 -

125 3.17 1.77

130 3.10 1.59

135 3.03 1.41

140 2.96 1.22

145 2.90 1.03

150 2.83 0.79

155 2.75 0.53

160 2.67 0.34

165 2.60 -0.11

170 2.53 -0.51

175 2.45 -1.20

180 2.38 -

185 2.28

190 2.21

195 2.13

200 2.05

205 1.96

210 1.87

215 1.79

220 1.69

225 1.61

230 1.53
235 1.44

240 1.31

245 1.19

250 1.10

255 0.99

260 0.88

265 0.74

270 0.64

275 0.47

280 0.34

285 0.26

290 0.10

295 -0.11

300 -0.36

305 -0.51

310 -0.92

315 -1.61

320 -2.30

325 -
 ln (C*-CL) for agitation speed, (rpm) against
6.00
time, (s)
ln (C* - CL) @ 200 rpm
ln (C* - CL) for agitation speed (rpm)

5.00
ln (C* - CL) @ 400 rpm
4.00
ln (C* - CL) @ 600 rpm
3.00
ln (C* - CL) @ 800 rpm
2.00 ln (C* - CL) @ 1000 rpm
1.00
0.00 y = -0.0171x + 5.1407 (200rpm)
0 50 100 150 200 250 300 350 -0.0292x + 5.1526 (400 rpm)
y =
-1.00 y = -0.0478x + 5.2738 (600 rpm)
-2.00 y = -0.062x + 5.2101 (800 rpm)
y = -0.072x + 5.3261 (1000 rpm)
-3.00
time (s)

Figure 2: The graph of ln (C* - CL) of agitation speed, rpm effect against time, s.

Table 3: The calculated value of kLa (h-1) for agitation speed effect given from the previous
graph.

Agitation speed (rpm) kLa value (h-1)

200 61.56
400 105.12
600 172.08
800 223.20
1000 259.20
 kLa value for agitation speed effect
300.00

250.00
kLa value (h-1)

200.00

150.00
kLa value for
100.00 agitation speed
effect
50.00

0.00
0 200 400 600 800 1000 1200
Agitation speed (rpm)

Figure 3: The kLa value (h-1) for each of the agitation speed value.
 B. The effect of aeration rate with constant of the agitation speed (400 rpm) and the
temperature (30˚C).
Table 4: The reading measured from the experiment for aeration rate effect.

Time (s) Aeration rate (L/min)

0.5 L/min 1.0 L/min 1.5 L/min 2.5 L/min

5 -1.70 -1.40 -1.70 -1.90

10 -1.80 -1.30 -1.40 -1.20

15 -1.70 0.58 0.57 1.48

20 -1.20 3.40 4.29 6.22

25 0.65 7.89 9.16 12.00

30 3.38 11.80 13.90 18.40

35 6.53 15.40 19.60 25.80

40 10.20 20.30 25.10 31.70

45 14.30 24.00 29.40 36.60

50 17.80 28.10 34.40 41.30

55 21.60 32.20 38.80 46.40

60 25.20 35.50 42.80 49.40

65 29.00 39.30 46.60 53.80

70 32.00 42.20 50.00 56.80

75 35.20 45.80 52.90 59.70

80 38.70 48.20 55.70 62.40

85 40.90 51.10 58.40 64.60

90 43.80 53.20 60.80 66.70

95 46.40 55.70 62.70 68.50

100 48.80 58.00 64.70 69.90

105 51.10 59.90 66.60 71.30

110 53.10 61.70 68.30 72.80

115 55.20 63.70 69.70 73.90

120 57.30 65.30 71.30 74.80

125 58.90 66.90 72.40 75.60

130 61.10 68.40 73.40 76.30

135 62.70 69.60 74.80 77.10

140 64.20 71.10 75.50 77.80

145 65.90 72.30 76.30 78.30

150 67.20 73.30 77.20 78.80

155 68.70 74.30 78.00 79.20

160 69.80 75.30 78.50 79.70

165 71.30 76.20 79.00 80.10

170 72.40 77.20 79.80 80.40

175 73.60 77.90 80.80 80.80

180 74.60 78.60 80.90 81.00

185 75.60 79.20 81.10 81.10

190 76.80 80.00 81.40 81.20

195 77.50 80.80 81.80 81.40

200 78.30 81.10 82.10 81.60

205 79.40 81.50 82.70 81.80

210 80.10 82.10 82.70 81.90

215 80.90 82.70 83.00 82.00

220 81.40 83.00 83.20 82.20

225 82.10 83.50 83.50 82.40

230 82.70 83.90 83.80 82.60

235 83.40 84.10 83.90 82.70

240 84.00 84.50 84.00 82.80

245 84.50 84.80 84.30 82.90

250 85.00 85.10 84.50 83.00

255 85.60 85.60 84.50 83.00

260 86.00 85.70 84.70 83.10

265 86.60 86.00 84.80 83.20

270 86.80 86.20 84.90 83.20

275 87.20 86.50 85.00 83.30

280 87.70 86.70 85.10 83.30

285 88.20 86.80 85.30 83.40

290 88.50 87.00 85.40 83.50

295 88.80 87.00 85.50 83.60

300 89.10 87.30 85.60 83.70

305 89.50 87.50 85.60 83.70

310 89.80 87.60 85.80 83.80

315 90.00 87.70 86.00 84.10

320 90.30 87.80 86.10

325 90.50 88.10 86.20

330 90.70 88.20 86.30

335 91.10 88.40 86.40

340 91.30 88.40 86.60

345 91.40 88.50 86.70

350 91.70 88.60

355 92.00 88.70

360 92.20 88.80

365 92.30 88.80

370 92.40 88.90

375 92.50 88.90

380 92.60 89.00

385 92.70 89.10

390 92.80 89.20

395 93.00 89.20

400 93.20 89.20

405 93.30 89.30

410 93.40 89.30

415 93.40 89.40

420 93.60 89.40

425 93.70 89.50

430 93.70 89.50

435 94.00 89.60

440 94.00 89.60

445 94.10 89.60

450 94.10 89.70

455 94.20 89.70

460 94.40 89.70

465 94.40 89.70

470 94.40 89.70

475 94.50

480 94.60
485 94.60

490 94.70

495 94.70

500 94.80

505 94.80

510 94.80

515 94.90

520 94.90

525 94.90

530 95.00

535 95.10

540 95.10

545 95.10

550 95.20

555 95.20

560 95.20

565 95.20

570 95.20
Table 5: The value calculated to plot the graph of ln (C*-CL) for aeration rate (L/min) against
time.

ln (C* - CL) ln (C* - CL) ln (C* - CL) ln (C* - CL)

Time (s)
at 0.5 L/min at 1.0 L/min at 1.5 L/min at 2.5 L/min
5 4.57 4.51 4.48 4.45

10 4.57 4.51 4.48 4.45

15 4.57 4.49 4.46 4.41

20 4.57 4.46 4.41 4.36

25 4.55 4.40 4.35 4.28

30 4.52 4.36 4.29 4.19

35 4.48 4.31 4.21 4.07

40 4.44 4.24 4.12 3.96

45 4.39 4.19 4.05 3.86

50 4.35 4.12 3.96 3.76

55 4.30 4.05 3.87 3.63

60 4.25 3.99 3.78 3.55

65 4.19 3.92 3.69 3.41

70 4.15 3.86 3.60 3.31

75 4.09 3.78 3.52 3.19

80 4.03 3.73 3.43 3.08

85 3.99 3.65 3.34 2.97

90 3.94 3.60 3.25 2.86

95 3.89 3.53 3.18 2.75

100 3.84 3.46 3.09 2.65

105 3.79 3.39 3.00 2.55

110 3.74 3.33 2.91 2.42

115 3.69 3.26 2.83 2.32

120 3.63 3.19 2.73 2.23

125 3.59 3.13 2.66 2.14

130 3.53 3.06 2.59 2.05

135 3.48 3.00 2.48 1.95

140 3.43 2.92 2.42 1.84

145 3.38 2.86 2.34 1.76

150 3.33 2.80 2.25 1.67

155 3.28 2.73 2.16 1.59

160 3.23 2.67 2.10 1.48

165 3.17 2.60 2.04 1.39

170 3.13 2.53 1.93 1.31

175 3.07 2.47 1.77 1.19

180 3.03 2.41 1.76 1.13

185 2.98 2.35 1.72 1.10

190 2.91 2.27 1.67 1.06

195 2.87 2.19 1.59 0.99

200 2.83 2.15 1.53 0.92

205 2.76 2.10 1.39 0.83

210 2.71 2.03 1.39 0.79

215 2.66 1.95 1.31 0.74

220 2.62 1.90 1.25 0.64

225 2.57 1.82 1.16 0.53

230 2.53 1.76 1.06 0.41

235 2.47 1.72 1.03 0.34

240 2.42 1.65 0.99 0.26

245 2.37 1.59 0.88 0.18

250 2.32 1.53 0.79 0.10

255 2.26 1.41 0.79 0.10

260 2.22 1.39 0.69 0.00

265 2.15 1.31 0.64 -0.11

270 2.13 1.25 0.59 -0.11

275 2.08 1.16 0.53 -0.22

280 2.01 1.10 0.47 -0.22

285 1.95 1.06 0.34 -0.36

290 1.90 0.99 0.26 -0.51

295 1.86 0.99 0.18 -0.69

300 1.81 0.88 0.10 -0.92

305 1.74 0.79 0.10 -0.92

310 1.69 0.74 -0.11 -1.20

315 1.65 0.69 -0.36 -

320 1.59 0.64 -0.51

325 1.55 0.47 -0.69

330 1.50 0.41 -0.92

335 1.41 0.26 -1.20

340 1.36 0.26 -2.30

345 1.34 0.18 -

350 1.25 0.10

355 1.16 0.00

360 1.10 -0.11

365 1.06 -0.11

370 1.03 -0.22

375 0.99 -0.22

380 0.96 -0.36

385 0.92 -0.51

390 0.88 -0.69

395 0.79 -0.69

400 0.69 -0.69

405 0.64 -0.92

410 0.59 -0.92

415 0.59 -1.20

420 0.47 -1.20

425 0.41 -1.61

430 0.41 -1.61

435 0.18 -2.30

440 0.18 -2.30

445 0.10 -2.30

450 0.10 -

455 0.00 -

460 -0.22 -

465 -0.22 -

470 -0.22 -

475 -0.36

480 -0.51

485 -0.51
490 -0.69

495 -0.69

500 -0.92

505 -0.92

510 -0.92

515 -1.20

520 -1.20

525 -1.20

530 -1.61

535 -2.30

540 -2.30

545 -2.30

550 -

555 -

560 -

565 -

570 -
 ln (C*-CL) for aeration rate, (L/min) against
time, (s)
6.00
5.00
ln (C*-CL) for aeration rate (L/min)

4.00
3.00
ln (C*-CL) @ 0.5 L/min
2.00
ln (C*-CL) @ 1.0 L/min
1.00
ln (C*-CL) @ 1.5 L/min
0.00
ln (C*-CL) @ 2.5 L/min
0 100 200 300 400 500 600
-1.00
y = -0.0114x + 5.0528 ( 0.5 L/min )
-2.00 y = -0.0142x + 4.9189 ( 1.0 L/min )
-3.00 y = -0.0162x + 4.7422 ( 1.5 L/min )
time (s) y = -0.0179x + 4.5323 ( 2.5 L/min )

Figure 4: The graph of ln (C* - CL) of aeration rate, L/min effect against time, s.

Table 6: The calculated value of kLa (h-1) for aeration rate effect given from the previous
graph.

Aeration rate (L/min) kLa value (h-1)

0.50 41.04
1.00 51.12
1.50 58.32
2.50 64.44
 kLa value of the aeration rate effect
70.00

60.00

50.00
kLa value (h-1)

40.00

30.00 kLa value of the

aeration rate
20.00 effect

10.00

0.00
0.00 0.50 1.00 1.50 2.00 2.50 3.00
Aeration rate (L/min)

Figure 5: The kLa value (h-1) for each of the aeration rate value.
 C. The effect of temperature with constant value of the agitation speed (400 rpm)
and the aeration rate (2 L/min).
Table 7: The reading measured from the experiment for the temperature effect.

Temperature (˚C)
Time (s) 30˚C 35˚C 40˚C 45˚C 50˚C
45.00 11.70 - - -
0
50.50 11.90 7.80 0.00 -0.10
5
55.90 13.40 15.90 2.77 2.46
10
61.00 17.70 24.20 9.97 8.60
15
65.50 24.00 31.50 17.70 19.80
20
69.60 31.10 40.70 28.10 31.80
25
73.30 39.10 47.80 37.50 43.00
30
77.20 46.30 55.50 46.50 52.80
35
79.50 52.70 61.80 55.30 62.50
40
82.40 58.90 68.30 63.30 70.20
45
84.60 64.50 72.90 68.90 77.70
50
86.30 69.30 77.70 75.20 84.50
55
88.10 73.50 81.40 79.90 89.30
60
89.80 77.50 85.40 84.30 93.90
65
91.20 81.00 87.90 88.00 97.50
70
92.40 84.00 90.90 91.50 100.00
75
93.50 86.70 93.00 94.20
80
94.60 88.80 95.20 96.90
85
95.30 90.80 96.70 98.60
90
96.10 92.40 98.40 100.00
95
96.80 93.90 99.50
100
97.40 95.20 100.00
105
 97.80 96.30
110
98.30 97.50
115
98.70 98.30
120
99.00 99.00
125
99.30 99.70
130
99.50 100.00
135
99.80
140
100.00
145

Table 8: The value calculated to plot the graph of ln (C*-CL) for temperature (˚C) against
time.

ln (C* - CL) ln (C* - CL) ln (C* - CL) ln (C* - CL) ln (C* - CL)
Time (s)
at 30˚C at 35˚C at 40˚C at 45˚C at 50˚C
4.01 4.48
0
5 3.90 4.48 4.52 4.61 4.61

10 3.79 4.46 4.43 4.58 4.58

15 3.66 4.41 4.33 4.50 4.52

20 3.54 4.33 4.23 4.41 4.38

25 3.41 4.23 4.08 4.28 4.22

30 3.28 4.11 3.96 4.14 4.04

35 3.13 3.98 3.80 3.98 3.85

40 3.02 3.86 3.64 3.80 3.62

45 2.87 3.72 3.46 3.60 3.39

50 2.73 3.57 3.30 3.44 3.10

55 2.62 3.42 3.10 3.21 2.74

60 2.48 3.28 2.92 3.00 2.37

65 2.32 3.11 2.68 2.75 1.81

70 2.17 2.94 2.49 2.48 0.92

75 2.03 2.77 2.21 2.14 -

80 1.87 2.59 1.95 1.76

85 1.69 2.42 1.57 1.13

90 1.55 2.22 1.19 0.34

95 1.36 2.03 0.47 -

100 1.16 1.81 -0.69

105 0.96 1.57 -

110 0.79 1.31

115 0.53 0.92

120 0.26 0.53

125 0.00 0.00

130 -0.36 -1.20

135 -0.69 -

140 -1.61

145 -
 ln (C*-CL) for temperature, (˚C) against
6.00 time, (s)
ln (C* - CL) @ T = 30
5.00
ln (C* - CL) @ T = 35
ln (C* - CL) for temperature (˚C)

4.00 ln (C* - CL) @ T = 40

ln (C* - CL) @ T = 45
3.00
ln (C* - CL) @ T = 50
2.00

1.00
y = -0.0366x + 5.1725 ( T = 35˚C)
0.00 y = -0.0345x + 4.3614 ( T = 30˚C )
0.00 20.00 40.00 60.00 80.00 100.00 120.00 140.00 160.00
-1.00 y = -0.0456x + 5.274 ( T = 40˚C)
y = -0.0449x + 5.3622 ( T = 45˚C)
-2.00 time (s) y = -0.0514x + 5.1123 ( T = 50˚C)

Figure 6: The graph of ln (C* - CL) of temperature, ˚C effect against time, s.

Table 9: The calculated value of kLa (h-1) for temperature effect given from the previous
graph.

Temperature (˚C) kLa value (h-1)

30 124.2
35 131.76
40 164.16
45 161.64
50 185.04
 kLa value of the temperature effect
200.00
180.00
kLa value for temperature (h-1)

160.00
140.00
120.00
100.00 kLa value of the
80.00 temperature
60.00 effect
40.00
20.00
0.00
0.00 10.00 20.00 30.00 40.00 50.00 60.00
Temperature (˚C)

Figure 7: The kLa value (h-1) for each of the temperature value.

SAMPLE CALLCULATION

In experiment 2, for aeration rate effect on the bioreactor experiment:

 For 0.5 L/min of aeration rate;

The C* = 95.2 at 550 s, CL = -1.7 at 5 s
The y-axis value is calculated by (Equation 1) as in the theory section:
ln (C* - CL) = ln [95.2 – (-1.7)]
ln (C* - CL) = 4.574
The value of 4.574 is then drawn into a graph against time for each time until it
reaches its maximum value as shown in the Figure 3 above.

From the graph plotted of the ln (C* - CL) against time, the equation of the best fit for
each rate was calculated as below:
Since y = mx + c
So, y = -0.0114x + 5.0528
Thus, the slope, m for the reaction is = -0.0114
From the theory section for (Equation 2), it is understood that the slope of the graph is
equal to the kLa value of that parameter at that condition.
Hence, kLa value for the 0.5 L/min of the aeration rate effect is = 0.0114 s-1.

The second graph is plotted based on the kLa value measured before against each
parameter as shown in the Figure 4.
Before plotting the graph, the value is converted into h-1 unit first, as shown below:
kLa (at 0.5 L/min) = 0.0114 s-1 x 60 s x 60 min
kLa (at 0.5 L/min) = 41.04 h-1.

The value of 41.04 h-1 is then plotted against the each of the aeration rate
experimented as shown in the Figure 4. The explanation of the graph will be
discussed in the Discussion section.
DISCUSSIONS

Based on the results tabulated and plotted above, it is generally concluded that the
higher the values of the parameters, the higher the values of the mass transfer coefficient, kLa.
For the first experiment which is the effect of the difference reading of the agitation speed in
rpm, it showed that the agitation speed of 1000 rpm reactions gives the faster reaction to
reached 100% value of pO2 reading compared to 200 rpm, 400 rpm, 600 rpm and 800 rpm. It
took only 80 s to achieve that value, while 800 rpm reaction took about 90 s, 600 rpm took
about 120 s, 400 rpm took about 180 s and finally the 200 rpm reaction took about 325 s to
reach 100%. From the value recorded for every 5 seconds, the difference between the
maximum concentration, C* and the value of each concentration for each time is calculated
thus measuring the value of the ln (C*-CL) to plot the first graph.

From Figure 2, it clearly shown that the 1000 rpm reaction gave the steeper line
compared to another speed due to the time taken for it achieving 100% much faster than
another. For that reason, the slope of that line much higher than another which is – 0.072. The
negative sign indicate that the graph has negative slope and does not effect on the real value
of the mass transfer coefficient. Then, the slope is taken as the value of the coefficient and
converted it in the unit of per hour (h-1). The 1000 rpm reaction gave about 259.2 h-1 for the
mass transfer coefficient value, while 800 rpm about 223.2 h-1, 600 rpm about 172.08 h-1, 400
rpm about 105.12 h-1, and 200 rpm about 61.56 h-1.

The graph plotted in the Figure 3 proved that the higher the agitation speed in rpm,
the higher the mass transfer coefficient. This is due to the agitation speed affect the problems
on the bubble size, bubble retention time and the mixing of the solution. The 1000 rpm
reaction efficiently breaking down the air bubbles since it has higher sectional area. Then, it
will affecting the superficial area of the bubbles than enhanced the transfer rate of the
oxygen. To that aim, the mass transfer coefficient for this reaction is much higher than
another speed while the lower speed does not have enough traps to hold up the air bubbles
and make the superficial area of the bubbles decreasing, thus lowering the value of the
transfer coefficient (Karimi, 2013).

Next is the effect of the aeration rate on the mass transfer coefficient of a reaction.
Based on the results tabulated and plotted above, it is generally concluded that the higher the
values of the parameters, the higher the values of the mass transfer coefficient, k La. It showed
that the aeration rate of 2.5 L/min reactions gives the faster reaction to reached 100% value of
pO2 reading compared to 0.5 L/min, 1.0 L/min and 1.5 L/min. It took only 315 s to achieve
that value, while 1.5 L/min reaction took about 345 s, 1.0 L/min took about 470 s and 0.5
L/min took about 550 s to reach 100%. From the value recorded for every 5 seconds, the
difference between the maximum concentration, C* and the value of each concentration for
each time is calculated thus measuring the value of the ln (C*-CL) to plot the first graph.

From Figure 4, it clearly shown that the 2.5 L/min reaction gave the steeper line
compared to another speed due to the time taken for it achieving 100% much faster than
another. For that reason, the slope of that line much higher than another which is – 0.0179.
The negative sign indicate that the graph has negative slope and does not effect on the real
value of the mass transfer coefficient. Then, the slope is taken as the value of the coefficient
and converted it in the unit of per hour (h-1). The 2.5 L/min reaction gave about 64.44 h-1 of
the mass transfer coefficient value meanwhile 1.5 L/min about 58.32 h-1, 1.0 L/min about
51.12 h-1 and 0.5 L/min about 41.04 h-1.

The graph plotted in the Figure 5 proved that the higher the aeration rates in L/min,
the higher the mass transfer coefficient. This is due to the aeration reaction will produced
small bubbles with uniform size of distribution and will lead to the increasing of the mass
transfer area between the gas and the liquid solution. This increasing mass transfer area is the
major reason for the transfer rate being increased as the aeration supplied increased in L/min
(Painmanakul, 2009).

Lastly is the effect of the temperature on the mass transfer coefficient of a reaction.
Based on the results tabulated and plotted above, it is generally concluded that the higher the
values of the parameters, the higher the values of the mass transfer coefficient, k La. It showed
that the temperature of 50˚C reactions gives the faster reaction to reached 100% value of pO2
reading compared to 30˚C, 35˚C, 40˚C and 45˚C. It took only 75 s to achieve that value,
while 45˚C reaction took about 95 s, 40˚C took about 105 s, 35˚C took about 135 s and 30˚C
took about 145 s to reach 100%. From the value recorded for every 5 seconds, the difference
between the maximum concentration, C* and the value of each concentration for each time is
calculated thus measuring the value of the ln (C*-CL) to plot the first graph.

From Figure 6, it clearly shown that the 50˚C reaction gave the steeper line compared
to another speed due to the time taken for it achieving 100% much faster than another. For
that reason, the slope of that line much higher than another which is – 0.0514. The negative
sign indicate that the graph has negative slope and does not effect on the real value of the
mass transfer coefficient. Then, the slope is taken as the value of the coefficient and
converted it in the unit of per hour (h-1). The 50˚C reaction gave about 185.04 h-1 of the mass
transfer coefficient value meanwhile 45˚C about 161.64 h-1, 40˚C about 164.16 h-1, 35˚C
about 131.76 h-1 and 30˚C about 124.20 h-1.

The graph plotted in the Figure 7 proved that the higher the temperature in ˚C, the
higher the mass transfer coefficient. This is due to the higher temperature caused the
concentration of the dissolved oxygen to be reduced. Thus, the mass transfer driving force,
which is (C*-CL) also being decreasing. This can be best explained by understanding that
higher temperature lead to the increasing of the air bubbles and increasing in the diffusivity
of the oxygen in the liquid film. After all, the mass transfer coefficient also being increased
(Ahmad, September 2017 - January 2018).
CONCLUSION

To be concluded the increasing in the agitation speed lead to the increasing in the
mass transfer coefficient. Same goes with the aeration rate effect on the transfer coefficient.
The higher the aeration rate supplied into the bioreactor, the higher the value of the mass
transfer coefficient. Lastly, the higher the temperature of the bioprocess system gives the
highest mass transfer coefficient of that reaction. This is because the transfer rate at the high
temperature much faster than the lower temperature. However, there is slightly little error in
this temperature experiment, where at the temperature 45˚C there is a little decreasing in
value due to the error in conducting the experiment. Theoretically, the highest value of mass
transfer coefficient is affected by the highest agitation speed of the bioreactor’s impeller,
highest aeration rate being supplied into the reactor as well as the highest temperature being
supplied through the temperature probe.

RECOMMENDATIONS

REFERENCES

Ahmad, I. N. (September 2017 - January 2018). Transport Processes in Stirred Tank

Bioreactors. Retrieved November, 2017

CliffNotes. (2016). The Scope of Biochemistry. Retrieved December, 2017, from Types of
Biochemical Reactions: https://www.cliffsnotes.com/study-
guides/biology/biochemistry-i/the-scope-of-biochemistry/types-of-biochemical-
reactions

Foundation, C.-1. (2017). cK-12. Retrieved December, 2017, from Types of Biochemical
Reactions: https://www.ck12.org/biology/types-of-biochemical-
reactions/lesson/Types-of-Biochemical-Reactions-BIO/

Kane, J. (1 March, 2012). BioProcess International. Retrieved December, 2017, from

Measuring kLa for Better Bioreactor Performance:
 http://www.bioprocessintl.com/upstream-processing/bioreactors/measuring-kla-for-
better-bioreactor-performance-328029/

Karimi, A. (7 January, 2013). US National Library of Medicine National Institutes of Health.

Retrieved December, 2017, from Oxygen mass transfer in a stirred tank bioreactor
using different impeller configurations for environmental purposes:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561095/

Painmanakul, P. (November, 2009). THEORETICAL PREDICTION OF VOLUMETRIC

MASS TRANSFER COEFFICIENT (kLa) FOR DESIGNING AN AERATION
TANK. Engineering Journal, 13(3), 14. doi:10.4186/ej.2009.13.3.13
APPENDICES

Figure 8: The nitrogen gas tank.

Figure 9: The oxygen gas supplier motor.