FACULTY OF CHEMICAL ENGINEERING

NAME : JAY MAC MILLAN BIN AZMAN

STUDENT ID : 2019582099
GROUP : EH2205B GROUP 1
EXPERIMENT : GROWTH STUDY OF MICROORGANISM IN BIOREACTOR
USING DIFFERENT STRATEGY
DATE PERFORMED : 13HB MAY 2020 WEEK 9
SEMESTER :5
PROGRAMME / CODE : REACTION ENGINEERING LABORATORY (CHE506)
LECTURER : DR. SHARMEELA MATALI

No. Title Allocated Marks (%) Marks

1 Abstract
2 Introduction
3 Objective
4 Theory
5 Materials & Apparatus
6 Methodology
7 Result
8 Calculation
9 Discussion
10 Conclusion
11 Recommendation
12 References
13 Appendices
TOTAL MARKS 100

Remarks:

Checked by:
TABLE OF CONTENTS

1.0 Abstract 1
2.0 Introduction 2-3
3.0 Objectives 4
4.0 Theory 5-7
5.0 Materials & Apparatus 8
6.0 Methodology 9-10
7.0 Results 11-21
8.0 Calculation 22
9.0 Discussion 23-24
10.0 Conclusion 25
11.0 Recommendation 26
12.0 References 27
13.0 Appendices 28-30
1.0 ABSTRACT
.

For this experiment, MINIFORS bioreactor is used to demonstrate its fundamental and working
principle and to achieve the experiment objectives. The main objective of the experiment to
investigate the effect of bioreactor parameters [agitation rate (RPM), the aeration rate (L/min)
and the temperature(°C)] on the dissolved oxygen concentration, CL (%). The method used is
static gassing out method. The volumetric mass transfer coefficient at agitation rate of 100
RPM, 200 RPM, 300 RPM, 400RPM and 500 RPM are 0.0078 s-1, 0.0117 s-1, 0.0241 s-1, 0.0341
s-1 and 0.0294 s-1, respectively. The greater the agitation rate magnitude, the greater the value
of the mass transfer coefficient. The volumetric mass transfer coefficient at aeration rate of 1
L/min, 2 L/min and 2.5 L/min are 0.0086 s-1, 0.0241 s-1 and 0.0472 s-1, respectively. The higher
the aeration rate magnitude, the greater the value of the volumetric mass transfer coefficient.
The volumetric mass transfer coefficient at temperature of 30°C, 35°C and 40°C are 0.0103 s-
1
, 0.0241 s-1, and 0.0472 s-1, respectively. The higher the temperature, the higher the magnitude
of the volumetric mass transfer coefficient. Comparing all three operating parameters, the most
substantial one is the agitation, followed by aeration and temperature, respectively. The
objectives are successfully obtained therefore the experiment is successfully done.

Page | 1
2.0 INTRODUCTION

Oxygen transfer in aerobic bioprocesses is crucial and any lack of oxygen massively affects
the process performance. Hence, oxygen mass transfer is one of the most significant
phenomena in the design and operation of mixing-sparging equipment for bioreactors
(Moutafchieva, 2013). Most of the biochemical processes require oxygen to yield the output.
Thus, maintaining appropriate concentration of dissolved oxygen, DO in the bioreactor is
crucial for the reactor efficiency. Hence, the reaction can be initiate, and the desired product
can be yield. Thus, the rate of transfer of oxygen between the two phases, gaseous and the
reaction liquid is investigated by determining the volumetric mass transfer coefficient, kLa in a
stirred tank with bubble aeration. The method used for the experiment is the gassing out method
in the absence of bacteria or cell. Agitation, aeration, and temperature are the three parameters
that affect the capability of a reactor’s oxygen mass transfer chosen for the experiment. To
understand it, MINIFORS bioreactor has been designed to identify the effect of parameters to
the bioreactor.

Page | 2
 Figure 1: The Phases of Bacteria Cell Growth

Table 1: Phases of Bacteria Cell Growth

Phase I The cell concentration shows a little increasing pattern. The cells are adjusting to
Lag Phase their new environment, synthesizing enzymes, and getting ready to begin
reproducing.
Phase II The rate of growth of the cell is proportional to the cell concentration. At this
Log Growth phase, all the enzyme’s pathways for metabolizing the substrate are in place.
Thus, the cells are dividing at maximum rate.
Phase III The net growth rate is zero since the cells reached the minimum biological space.
Stationary Phase
Phase IV Decrease in live cell concentration.
Death Phase

Page | 3
3.0 OBJECTIVES

The main objective of the experiment is to investigate the effect of bioreactor parameters
[agitation rate (RPM), the aeration rate (L/min) and the temperature(°C)] on the dissolved
oxygen concentration, CL (%).

There were several sub-objectives in conducting the experiment:

1) To identify the driving force, (C* - CL) of the stirred tank reactor.
2) To plot the graph of ln(C* - CL) against bioreactor parameters
3) To calculate the volumetric mass transfer coefficient (kLa) of the stirred tank reactor.
4) To use the gassing out method for the experiment.

Page | 4
4.0 THEORY

MINIFORS (Stirred tank bioreactor) reactors most generally utilized for culturing of biological
agents such as cells, enzymes, or antibodies. They are contactors where the well mixed among
phases is obtained mainly by internal mechanical agitation (Garcia-Ochoa, 2011). The gassing
– out method is used in the experiment to determine the volumetric mass transfer coefficient,
kLa in the bioreactor using water as the only simulating media. Nitrogen gas is used to gassing
out the liquid so that the oxygen concentration of the solution is lowered. Thus, making the
solution scrubbed free of oxygen.

The increase in dissolved oxygen is monitored after the deoxygenated liquid is aerated and
agitated. This method utilizes the graphical technique to determine the experimental values of
volumetric mass transfer coefficient, kLa. The oxygen transfer rate decreases as the driving
force decreases. The oxygen transfer rate will be equal to the slope of tangent to the curve of
values of dissolved oxygen concentration against time of aeration. (Organisms, 2018).

Dynamic Gassing Out Method

Oxygen Transfer Rate (OTR) is the rate at which oxygen is transferred into solution.

𝐎𝐓𝐑 = 𝐤𝐋𝐀(𝐂∗ − 𝐂𝐋)

Where,

kL = oxygen transfer coefficient (cm/h)

A = gas – liquid interfacial area (cm2/cm3)

kLA = Volumetric oxygen transfer coefficient (h-1)

C* = Saturated dissolved oxygen concentration (mg/L)

CL = Actual dissolved ocygen concentration in the broth (mg/L)

OTR = oxygen transfer rate (mg O2 / L.h)

Page | 5
Oxygen Uptake Rate (OUR) is the rate at which bacteria or other microorganisms consume

oxygen.

𝐎𝐔𝐑 = 𝐪𝐎𝟐𝐗

Where,

qO2 = specific rate of oxygen consumption (mmol O2/gdw.h)

X = bacteria concentration (gdw/L)

gdw = gram dry weight of cells

Substituting the OUR and OTR equation yield the following equation,

𝒅𝑪𝑳
= 𝒌𝑳 𝒂(𝑪∗ − 𝑪𝑳 ) − 𝒒𝑶𝟐 𝑿
𝒅𝒕

The plot of CL against dCL/dt + qO2X , the slope equal to -1/kLa.

Figure 2: CL against dCL/dt + qO2X

Page | 6
Static Gassing Out Method
The increased in dissolved oxygen concentration is given by the following equation,
𝒅𝑪𝑳
= 𝒌𝑳 𝒂(𝑪∗ − 𝑪𝑳 )
𝒅𝒕
Integrating the equation yield to the following equation,
∫ (𝐂∗𝐝− 𝐂𝐋𝐂𝐋) = ∫ 𝐤𝐋𝐀 𝐝𝐭
𝐥𝐧(𝐂∗ − 𝐂𝐋) = 𝐤𝐋𝐀. 𝐭
The plot of the ln (C* - CL) against time, the slope equals to -kLa

Figure 3: ln(C*-CL) versus time

Page | 7
5.0 MATERIAL & APPARATUS

5.1 Materials
1. Distilled water
2. Nitrogen gas (purged gas)

5.2 Apparatus
1. MINIFORS bioreactor
2. Stop watch
3. (HI-BLOW HP 80) Linear air pump aerator

Figure 4: MINIFORS bioreactor

Page | 8
6.0 METHODOLOGY

6.1 Start-up Procedures.

1. The reactor is turned on and leaved for around 30 minutes to stabilize the oxygen
probe.
2. Air line of air compressor is connected to the reactor.
3. All parameters are ensured on “ON” mode.

6.2 Oxygen Probe (PO2) Calibration

1. At reactor display, once PO2 mode is ON, the calibration button is (the one
underneath the “F” button) pressed until “cAL’ is displayed on the display.
2. 2 points (i.e 0 and 100%) are obtained by pressing “+” button.
3. Gas valve is opened. Rotameter us used to set the flowrate to the desired value.
4. Air is purged out from the vessel with nitrogen.
5. Air line is replaced with nitrogen at the back of the reactor. Air compressor is
switched off once the air line has been pulled out.
6. Once the reading shows ‘0’ “ON/OFF” pressed once to accept the value.
7. For 2nd point (i.e 100% air), the nitrogen line is pulled out and replaced with air line.
8. Wait and once the value reach ‘100’, “ON/OFF” button is pressed once to accept the
value.
9. “ON/OFF” button pressed once again to END the calibration.

6.3 Effect of agitation rate (RPM)

1. The aeration rate parameter is set up at 2.0 L/min and the temperature parameter is set
up at 30°C.
2. The pump is then switched off.
3. The first agitation parameter is set up at 100 rpm and prepared for 2 points calibration.
The setting is done before the purging using nitrogen gas.
4. Then, nitrogen gas is purged on the system until the value of the partial pressure of
oxygen inside the system becomes 0%.
5. The nitrogen gas valve is then closed, and the tube attached to the equipment is
detached from the reactor. The pump is then switched back on allowing the aeration
of air inside the reactor.
6. The actual dissolved oxygen concentration, PO2 (%) is recorded every 5 seconds until
the reading reached to PO2 = 100%. The reading is stopped after three consecutive
reading yields to the same value
7. Step 3 until Step 6 is repeated for different agitation rate parameter value which are
200 RPM, 300 RPM and 400 RPM.

Page | 9
6.4 Effect of aeration (L/min)
1. The agitation rate parameter is set up at 300 RPM and the temperature parameter is
set up at 30°C.
2. The pump is then switched off.
3. The first aeration rate parameter is set up at 1 L/min and prepared for 2 points
calibration. The setting is done before the purging using nitrogen gas.
4. Then, nitrogen gas is purged on the system until the value of the partial pressure of
oxygen inside the system becomes 0%.
5. The nitrogen gas valve is then closed, and the tube attached to the equipment is
detached from the reactor. The pump is then switched back on allowing the aeration
of air inside the reactor.
6. The actual dissolved oxygen concentration, PO2 (%) is recorded every 5 seconds until
the reading reached to PO2 = 100%. The reading is stopped after three consecutive
reading yields to the same value
7. Step 3 until Step 6 is repeated for different aeration rate parameter value which are 2.0
L/min and 2.5 L/min.

6.5 Effect of temperature (°C)

1. The aeration rate parameter is set up at 2.0 L/min and the agitation parameter is set up
at 300 RPM.
2. The pump is then switched off.
3. The first temperature parameter is set up at 30°C and prepared for 2 points calibration.
The setting is done before the purging using nitrogen gas.
4. Then, nitrogen gas is purged on the system until the value of the partial pressure of
oxygen inside the system becomes 0%.
5. The nitrogen gas valve is then closed, and the tube attached to the equipment is
detached from the reactor. The pump is then switched back on allowing the aeration
of air inside the reactor.
6. The actual dissolved oxygen concentration, PO2 (%) is recorded every 5 seconds until
the reading reached to PO2 = 100%. The reading is stopped after three consecutive
reading yields to the same value
7. Step 3 until Step 6 is repeated for different agitation parameter value which are 35°C
and 40°C.

6.6 Shut-down Procedures.

1. All the parameters are switched off by pressing the ‘ON/OFF/’ button
2. MINIFORS bioreactor and pump are switched off.

Page | 10
 7.0 RESULTS

Table 2: Percentage of dissolved oxygen concentration, CL (%) at various agitation rate, 2 L/min aeration rate
and temperature of 30°C.

Agitation
(RPM) 100 200 300 400 100 200 300 400 500 100 200 300 400 500

Time, Actual dissolved oxygen C* - CL Ln (C* - CL)

t (s) concentration, CL (%)
0 0.00 0.00 0.00 0.00 100 100 100 100 0 4.61 4.61 4.61 4.61 4.61
5 0.00 0.00 0.00 1.22 100 100 100 98.78 2.44 4.61 4.61 4.61 4.59 4.58
10 0.00 0.00 2.12 5.01 100 100 97.88 94.99 8.26 4.61 4.61 4.58 4.55 4.52
15 0.00 1.19 5.55 12.1 100 98.81 94.45 87.9 13.9 4.61 4.59 4.55 4.48 4.46
20 0.43 2.23 10.4 14.4 99.57 97.77 89.6 85.6 21.5 4.6 4.58 4.50 4.45 4.36
25 1.30 3.15 16.7 26.1 98.70 96.85 83.3 73.9 28.8 4.59 4.57 4.42 4.3 4.27
30 2.23 5.47 22.5 34.0 97.77 94.53 77.5 66 35.3 4.58 4.55 4.35 4.19 4.17
35 3.42 7.75 27.8 40.9 96.58 92.25 72.2 59.1 43.1 4.57 4.52 4.28 4.08 4.04
40 4.76 10.6 34.2 47.7 95.24 89.4 65.8 52.3 48.9 4.56 4.49 4.19 3.96 3.93
45 6.21 13.5 38.8 54.0 93.79 86.5 61.2 46 54.7 4.54 4.46 4.11 3.83 3.81
50 7.93 15.8 44.1 60.1 92.07 84.2 55.9 39.9 60 4.52 4.43 4.02 3.69 3.69
55 9.51 18.8 48.7 65.0 90.49 81.2 51.3 35 65.1 4.51 4.4 3.93 3.56 3.55
60 11.3 21.6 53.7 69.5 88.7 78.4 46.3 30.5 69.1 4.49 4.36 3.84 3.42 3.43
65 13.3 24.4 57.5 73.6 86.7 75.6 42.5 26.4 72.5 4.46 4.33 3.75 3.27 3.31
70 15.5 27.4 61.4 77.1 84.5 72.6 38.6 22.9 75.6 4.44 4.28 3.65 3.13 3.19
75 17.6 29.9 65.0 80.2 82.4 70.1 35 19.8 78.5 4.41 4.25 3.56 2.99 3.07
80 19.3 32.8 68.3 82.3 80.7 67.2 31.7 17.7 81.3 4.39 4.21 3.46 2.87 2.93
85 21.1 35.2 71.6 85.5 78.9 64.9 28.4 14.5 83.3 4.37 4.17 3.35 2.67 2.82
90 23.2 37.5 74.1 87.8 76.8 62.5 25.9 12.2 85.5 4.34 4.14 3.25 2.5 2.67
95 25.4 40.0 76.6 89.4 74.6 60 23.4 10.6 87.5 4.31 4.09 3.15 2.36 2.53
100 26.9 42.4 79.1 91.1 73.1 57.6 20.9 8.9 88.7 4.29 4.05 3.04 2.19 2.42
105 29.0 44.8 81.2 92.4 71 55.2 18.8 7.6 90.1 4.26 4.01 2.93 2.03 2.29
110 30.7 46.7 83.1 93.8 69.3 53.3 15.9 6.2 91.4 4.24 3.98 2.77 1.82 2.15
115 32.0 48.7 84.8 94.8 68 51.3 15.2 5.2 92.4 4.22 3.94 2.72 1.65 2.03
120 34.5 50.8 86.3 95.7 65.5 49.2 13.7 4.3 93.4 4.18 3.9 2.62 1.46 1.89
125 36.1 52.6 87.5 96.4 63.9 47.4 12.5 3.6 94.3 4.16 3.86 2.53 1.28 1.74
130 38.1 54.4 89.0 97.1 61.9 45.6 11 2.9 94.9 4.13 3.82 2.4 1.06 1.63
135 40.0 56.4 90.0 97.7 60 43.6 10 2.3 95.7 4.09 3.78 2.3 0.83 1.46
140 41.7 58.2 91.2 98.3 58.3 41.8 8.8 1.7 96.2 4.07 3.73 2.17 0.53 1.34
145 43.5 60.0 92.2 98.8 56.5 40 7.2 1.2 96.8 4.03 3.69 1.97 0.18 1.16
150 45.1 61.7 93.2 99.2 54.9 38.3 6.8 0.8 97.3 4.01 3.65 1.92 -0.22 0.99
155 46.8 63.3 93.9 99.6 53.2 36.7 6.1 0.4 97.7 3.97 3.6 1.81 -0.92 0.83
160 48.5 64.9 94.6 99.8 51.5 35.1 5.4 0.2 98.1 3.94 3.56 1.69 -1.61 0.64
165 50.1 66.3 95.2 100 49.9 33.7 4.8 0 98.5 3.91 3.52 1.57 - 0.41
170 51.6 67.4 95.9 - 48.4 32.6 4.1 - 98.8 3.88 3.48 1.41 - 0.18
175 53.0 68.7 96.5 - 47 31.3 3.5 - 99 3.85 3.44 1.25 - 0.00
180 54.3 70.1 96.9 - 45.7 29.9 3.1 - 99.3 3.82 3.4 1.13 - -0.36
185 55.8 71.2 97.4 - 44.2 28.8 2.6 - 99.6 3.79 3.36 0.96 - -0.92
190 57.4 72.5 97.8 - 42.6 27.5 2.1 - 99.8 3.75 3.31 0.74 - -1.61
195 58.6 73.4 98.2 - 41.4 26.6 1.8 - 99.8 3.72 3.28 0.59 - -1.61
200 59.8 74.5 98.6 - 40.2 25.5 1.4 - 100 3.69 3.24 0.34 - -
205 61.3 75.8 98.9 - 38.7 24.2 1.1 - - 3.66 3.19 0.1 - -

Page | 11
210 62.5 76.7 99.2 - 37.5 23.3 0.8 - - 3.62 3.15 -0.22 - -
215 63.8 77.8 99.4 - 36.2 22.2 0.6 - - 3.59 3.1 -0.51 - -
220 65.2 78.9 99.7 - 34.8 21.1 0.3 - - 3.55 3.05 -1.2 - -
225 66.3 80.1 99.8 - 34.7 19.9 0.2 - - 3.55 2.99 -1.61 - -
230 67.2 80.8 100 - 32.8 19.2 0 - - 3.49 2.95 - - -
235 68.1 81.7 - - 31.9 18.3 - - - 3.46 2.91 - - -
240 69.0 82.4 - - 31 17.6 - - - 3.43 2.87 - - -
245 69.7 83.1 - - 30.3 16.9 - - - 3.41 2.83 - - -
250 70.7 83.9 - - 29.3 16.1 - - - 3.38 2.78 - - -
255 71.7 84.8 - - 28.3 15.2 - - - 3.34 2.72 - - -
260 72.5 85.5 - - 27.5 14.5 - - - 3.31 2.67 - - -
265 73.4 86.1 - - 26.6 13.9 - - - 3.28 2.63 - - -
270 74.3 86.7 - - 25.7 13.3 - - - 3.25 2.59 - - -
275 75.1 87.4 - - 24.9 12.6 - - - 3.21 2.53 - - -
280 75.8 88.1 - - 24.2 11.9 - - - 3.19 2.48 - - -
285 76.6 88.9 - - 23.4 11.1 - - - 3.15 2.41 - - -
290 77.4 89.5 - - 22.6 10.5 - - - 3.12 2.35 - - -
295 78.4 90.0 - - 21.6 10 - - - 3.07 2.3 - - -
300 79.1 90.9 - - 20.9 9.1 - - - 3.04 2.21 - - -
305 80.0 91.3 - - 20 8.7 - - - 3 2.16 - - -
310 80.6 91.8 - - 19.4 8.2 - - - 2.97 2.1 - - -
315 81.2 92.3 - - 18.8 7.7 - - - 2.93 2.04 - - -
320 81.8 92.9 - - 18.2 7.1 - - - 2.9 1.96 - - -
325 82.3 93.5 - - 17.7 6.5 - - - 2.87 1.87 - - -
330 82.9 94.0 - - 17.1 6 - - - 2.84 1.79 - - -
335 83.4 94.3 - - 16.6 5.7 - - - 2.81 1.74 - - -
340 83.9 94.8 - - 16.1 5.2 - - - 2.78 1.65 - - -
345 84.4 95.1 - - 15.6 4.9 - - - 2.75 1.59 - - -
350 85.0 95.4 - - 15 4.6 - - - 2.71 1.53 - - -
355 85.7 95.7 - - 14.3 4.3 - - - 2.66 1.46 - - -
360 86.1 96.0 - - 13.9 4 - - - 2.63 1.39 - - -
365 86.4 96.3 - - 13.6 3.7 - - - 2.61 1.31 - - -
370 87.0 96.6 - - 13 3.4 - - - 2.56 1.22 - - -
375 87.3 96.8 - - 12.7 3.2 - - - 2.54 1.16 - - -
380 87.8 97.1 - - 12.2 2.9 - - - 2.5 1.06 - - -
385 88.2 97.3 - - 11.8 2.6 - - - 2.47 0.96 - - -
390 88.5 97.5 - - 11.5 2.5 - - - 2.44 0.92 - - -
395 88.8 98.1 - - 11.2 1.9 - - - 2.42 0.64 - - -
400 89.2 98.3 - - 10.8 1.7 - - - 2.38 0.53 - - -
405 89.6 98.5 - - 10.4 1.5 - - - 2.34 0.41 - - -
410 90.0 98.6 - - 10 1.4 - - - 2.3 0.34 - - -
415 90.4 98.8 - - 9.6 1.2 - - - 2.26 0.18 - - -
420 90.9 98.9 - - 9.1 1.1 - - - 2.21 0.1 - - -
425 91.2 99.1 - - 8.8 0.9 - - - 2.17 -0.11 - - -
430 91.6 99.2 - - 8.4 0.8 - - - 2.13 -0.22 - - -
435 91.9 99.3 - - 8.1 0.7 - - - 2.09 -0.36 - - -
440 92.2 99.4 - - 7.8 0.6 - - - 2.05 --0.51 - - -
445 92.5 99.6 - - 7.5 0.4 - - - 2.01 -0.92 - - -
450 92.7 99.7 - - 7.3 0.3 - - - 1.99 -1.2 - - -
455 92.9 99.8 - - 7.1 0.2 - - - 1.96 -1.61 - - -
460 93.2 99.9 - - 6.8 0.1 - - - 1.92 -2.3 - - -
465 93.4 100 - - 6.6 0 - - - 1.89 - - - -
470 93.7 - - - 6.3 - - - - 1.84 - - - -

Page | 12
475 93.8 - - - 6.2 - - - - 1.82 - - - -
480 94.0 - - - 6 - - - - 1.79 - - - -
485 94.3 - - - 5.7 - - - - 1.74 - - - -
490 94.5 - - - 5.5 - - - - 1.7 - - - -
495 94.8 - - - 5.2 - - - - 1.65 - - - -
500 94.9 - - - 5.1 - - - - 1.63 - - - -
505 95.0 - - - 5 - - - - 1.61 - - - -
510 95.2 - - - 4.8 - - - - 1.57 - - - -
515 95.3 - - - 4.7 - - - - 1.55 - - - -
520 95.5 - - - 4.5 - - - - 1.5 - - - -
525 95.7 - - - 4.3 - - - - 1.46 - - - -
530 95.9 - - - 4.1 - - - - 1.41 - - - -
535 96.1 - - - 3.9 - - - - 1.36 - - - -
540 96.3 - - - 3.7 - - - - 1,31 - - - -
545 96.4 - - - 3.6 - - - - 1.28 - - - -
550 96.5 - - - 3.5 - - - - 1.25 - - - -
555 96.6 - - - 3.4 - - - - 1.22 - - - -
560 96.8 - - - 3.2 - - - - 1.16 - - - -
565 96.9 - - - 3.1 - - - - 1.13 - - - -
570 97.0 - - - 3 - - - - 1.1 - - - -
575 97.2 - - - 2.8 - - - - 1.03 - - - -
580 97.3 - - - 2.7 - - - - 0,99 - - - -
585 97.4 - - - 2.6 - - - - 0.96 - - - -
590 97.5 - - - 2.5 - - - - 0.92 - - - -
600 97.6 - - - 2.4 - - - - 0.88 - - - -
605 97.7 - - - 2.3 - - - - 0.83 - - - -
610 97.8 - - - 2.2 - - - - 0.79 - - - -
615 98.0 - - - 2 - - - - 0.69 - - - -
620 98.1 - - - 1.9 - - - - 0.64 - - - -
625 98.2 - - - 1.8 - - - - 0.59 - - - -
630 98.4 - - - 1.6 - - - - 0.47 - - - -
635 98.5 - - - 1.5 - - - - 0.41 - - - -
640 98.6 - - - 1.4 - - - - 0.34 - - - -
645 98.7 - - - 1.3 - - - - 0.26 - - - -
650 98.8 - - - 1.2 - - - - 0.18 - - - -
655 98.9 - - - 1.1 - - - - 0.1 - - - -
660 99.0 - - - 1 - - - - 0 - - - -
665 99.1 - - - 0.9 - - - - -0.11 - - - -
670 99.2 - - - 0.8 - - - - -0.22 - - - -
675 99.3 - - - 0.7 - - - - -0.36 - - - -
680 99.4 - - - 0.6 - - - - -0.51 - - - -
685 99.5 - - - 0.5 - - - - -0.69 - - - -
690 99.6 - - - 0.4 - - - - -0.92 - - - -
695 99.7 - - - 0.3 - - - - -1.2 - - - -
700 99.0 - - 0.1 - - - - -1.3 - - - -
705 99.1 - - 0.9 - - - - -0.11 - - - -
710 99.4 - - 0.6 - - - - -0.51 - - - -
715 99.6 - - 0.4 - - - - -0.92 - - - -
720 99.8 - - 0.2 - - - - -1.61 - - - -
725 99.8 - - 0.2 - - - - -1.91 - - - -
730 99.9 - - 0.1 - - - - -1.3 - - - -
735 99.9 - - 0.1 - - - - -1.3 - - - -
740 99.9 - - 0.1 - - - - -1.3 - - - -

Page | 13
Table 3: Percentage of dissolved oxygen concentration, CL (%) at various aeration rate, 300 RPM agitation
rate and temperature of 30°C.

Aeration rate
(L/min) 1 2 2.5 1 2 2.5 1 2 2.5
Time,
t (s) Actual dissolved oxygen C* - CL Ln (C* - CL)
concentration, CL (%)
0 0 0 1.27 100.00 100.00 98.73 4.61 4.61 4.59
5 0 0 8.3 100.00 100.00 91.70 4.61 4.61 4.52
10 0.04 2.12 17.7 99.96 97.88 82.30 4.60 4.58 4.41
15 0.84 5.55 27.9 99.16 94.45 72.10 4.60 4.55 4.28
20 2.58 10.4 37.9 97.42 89.60 62.10 4.58 4.50 4.13
25 5.14 16.7 46.7 94.86 83.30 53.30 4.55 4.42 3.98
30 8.05 22.5 54.8 91.95 77.50 45.20 4.52 4.35 3.81
35 11.8 27.8 62.5 88.20 72.20 37.50 4.48 4.28 3.62
40 14.9 34.2 68.3 85.10 65.80 31.70 4.44 4.19 3.46
45 18.3 38.8 74 81.70 61.20 26.00 4.40 4.11 3.26
50 22.6 44.1 78.5 77.40 55.90 21.50 4.35 4.02 3.07
55 25.9 48.7 82.1 74.10 51.30 17.90 4.31 3.94 2.88
60 29.7 53.7 85.4 70.30 46.30 14.60 4.25 3.84 2.68
65 33.2 57.5 87.8 66.80 42.50 12.20 4.20 3.75 2.50
70 37.4 61.4 90 62.60 38.60 10.00 4.14 3.65 2.30
75 40.4 65 91.7 59.60 35.00 8.30 4.09 3.56 2.12
80 43.6 68.3 93.2 56.40 31.70 6.80 4.03 3.46 1.92
85 46.4 71.6 94.4 53.60 28.40 5.60 3.98 3.35 1.72
90 49.3 74.1 95.4 50.70 25.90 4.60 3.93 3.25 1.53
95 52 76.6 96.2 48.00 23.40 3.80 3.87 3.15 1.34
100 54.8 79.1 97.9 45.20 20.90 2.10 3.81 3.04 0.74
105 57.4 81.2 98.3 42.60 18.80 1.70 3.75 2.93 0.53
110 59.5 83.1 99.1 40.50 16.90 0.90 3.70 2.83 -0.11
115 61.5 84.8 99.4 38.50 15.20 0.60 3.65 2.72 -0.51
120 63.7 86.3 99.6 36.30 13.70 0.40 3.59 2.62 -0.92
125 65.9 87.5 99.7 34.10 12.50 0.30 3.53 2.53 -1.20
130 67.5 89 99.9 32.50 11.00 0.10 3.48 2.40 -2.30
135 69.1 90 100 30.90 10.00 0.00 3.43 2.30 -
140 70.8 91.2 - 29.20 8.80 - 3.37 2.17 -
145 72.6 92.2 - 27.40 7.80 - 3.31 2.05 -
150 74 93.2 - 26.00 6.80 - 3.26 1.92 -
155 75.3 93.9 - 24.70 6.10 - 3.21 1.81 -
160 76.6 94.6 - 23.40 5.40 - 3.15 1.69 -
165 78 95.2 - 22.00 4.80 - 3.09 1.57 -
170 79.2 95.9 - 20.80 4.10 - 3.03 1.41 -
175 80.3 96.5 - 19.70 3.50 - 2.98 1.25 -
180 81.3 96.9 - 18.70 3.10 - 2.93 1.13 -
185 82.5 97.4 - 17.50 2.60 - 2.86 0.96 -
190 83.3 97.8 - 16.70 2.20 - 2.82 0.79 -
195 84 98.2 - 16.00 1.80 - 2.77 0.59 -
200 84.8 98.6 - 15.20 1.40 - 2.72 0.34 -

Page | 14
205 85.7 98.9 - 14.30 1.10 - 2.66 0.10 -
210 86.2 99.2 - 13.80 0.80 - 2.62 -0.22 -
215 87 99.4 - 13.00 0.60 - 2.56 -0.51 -
220 87.5 99.7 - 12.50 0.30 - 2.53 -1.20 -
225 88.1 99.8 - 11.90 0.20 - 2.48 -1.61 -
230 88.6 100 - 11.40 0.00 - 2.43 - -
235 89.2 - - 10.80 - - 2.38 - -
240 89.6 - - 10.40 - - 2.34 - -
245 90.1 - - 9.90 - - 2.29 - -
250 90.6 - - 9.40 - - 2.24 - -
255 91 - - 9.00 - - 2.20 - -
260 91.8 - - 8.20 - - 2.10 - -
265 92.3 - - 7.70 - - 2.04 - -
270 92.6 - - 7.40 - - 2.00 - -
275 92.9 - - 7.10 - - 1.96 - -
280 93.1 - - 6.90 - - 1.93 - -
285 93.5 - - 6.50 - - 1.87 - -
290 93.7 - - 6.30 - - 1.84 - -
295 94 - - 6.00 - - 1.79 - -
300 94.2 - - 5.80 - - 1.76 - -
305 94.4 - - 5.60 - - 1.72 - -
310 94.7 - - 5.30 - - 1.67 - -
315 94.8 - - 5.20 - - 1.65 - -
320 95 - - 5.00 - - 1.61 - -
325 95.2 - - 4.80 - - 1.57 - -
330 95.3 - - 4.70 - - 1.55 - -
335 95.5 - - 4.50 - - 1.50 - -
340 95.7 - - 4.30 - - 1.46 - -
345 95.8 - - 4.20 - - 1.44 - -
350 95.9 - - 4.10 - - 1.41 - -
355 96 - - 4.00 - - 1.39 - -
360 96.2 - - 3.80 - - 1.34 - -
365 96.4 - - 3.60 - - 1.28 - -
370 96.5 - - 3.50 - - 1.25 - -
375 96.5 - - 3.50 - - 1.25 - -
380 96.5 - - 3.50 - - 1.25 - -
385 96.6 - - 3.40 - - 1.22 - -
390 96.7 - - 3.30 - - 1.19 - -
395 96.8 - - 3.20 - - 1.16 - -
400 96.9 - - 3.10 - - 1.13 - -
405 97 - - 3.00 - - 1.10 - -
410 97 - - 3.00 - - 1.10 - -
415 97.1 - - 2.90 - - 1.06 - -
420 97.2 - - 2.80 - - 1.03 - -
425 97.2 - - 2.80 - - 1.03 - -
430 97.4 - - 2.60 - - 0.96 - -
435 97.5 - - 2.50 - - 0.92 - -
440 97.5 - - 2.50 - - 0.92 - -
445 97.5 - - 2.50 - - 0.92 - -
450 97.5 - - 2.50 - - 0.92 - -

Page | 15
 455 97.6 - - 2.40 - - 0.88 - -
460 97.7 - - 2.30 - - 0.83 - -
465 97.7 - - 2.30 - - 0.83 - -
470 97.7 - - 2.30 - - 0.83 - -
475 97.7 - - 2.30 - - 0.83 - -
480 97.8 - - 2.20 - - 0.79 - -
485 97.8 - - 2.20 - - 0.79 - -
490 97.9 - - 2.10 - - 0.74 - -
495 97.9 - - 2.10 - - 0.74 - -

Table 4: Percentage of dissolved oxygen concentration, CL (%) at various temperature (°C), 300 rpm agitation
rate and aeration rate of 2 L/min.

Temperature
(°C) 30 35 40 30 35 40 30 35 40

Time, t Actual dissolved oxygen C* - CL Ln (C* - CL)

(s) concentration, CL (%)
0 0 0 1.27 100.00 100.00 98.73 4.61 4.61 4.59
5 0 0 8.3 100.00 100.00 91.70 4.61 4.61 4.52
10 0.04 2.12 17.7 99.96 97.88 82.30 4.60 4.58 4.41
15 0.84 5.55 27.9 99.16 94.45 72.10 4.60 4.55 4.28
20 2.58 10.4 37.9 97.42 89.60 62.10 4.58 4.50 4.13
25 5.14 16.7 46.7 94.86 83.30 53.30 4.55 4.42 3.98
30 8.05 22.5 54.8 91.95 77.50 45.20 4.52 4.35 3.81
35 11.8 27.8 62.5 88.20 72.20 37.50 4.48 4.28 3.62
40 14.9 34.2 68.3 85.10 65.80 31.70 4.44 4.19 3.46
45 18.3 38.8 74 81.70 61.20 26.00 4.40 4.11 3.26
50 22.6 44.1 78.5 77.40 55.90 21.50 4.35 4.02 3.07
55 25.9 48.7 82.1 74.10 51.30 17.90 4.31 3.94 2.88
60 29.7 53.7 85.4 70.30 46.30 14.60 4.25 3.84 2.68
65 33.2 57.5 87.8 66.80 42.50 12.20 4.20 3.75 2.50
70 37.4 61.4 90 62.60 38.60 10.00 4.14 3.65 2.30
75 40.4 65 91.7 59.60 35.00 8.30 4.09 3.56 2.12
80 43.6 68.3 93.2 56.40 31.70 6.80 4.03 3.46 1.92
85 46.4 71.6 94.4 53.60 28.40 5.60 3.98 3.35 1.72
90 49.3 74.1 95.4 50.70 25.90 4.60 3.93 3.25 1.53
95 52 76.6 96.2 48.00 23.40 3.80 3.87 3.15 1.34
100 54.8 79.1 97.9 45.20 20.90 2.10 3.81 3.04 0.74
105 57.4 81.2 98.3 42.60 18.80 1.70 3.75 2.93 0.53
110 59.5 83.1 99.1 40.50 16.90 0.90 3.70 2.83 -0.11
115 61.5 84.8 99.4 38.50 15.20 0.60 3.65 2.72 -0.51
120 63.7 86.3 99.6 36.30 13.70 0.40 3.59 2.62 -0.92
125 65.9 87.5 99.7 34.10 12.50 0.30 3.53 2.53 -1.20
130 67.5 89 99.9 32.50 11.00 0.10 3.48 2.40 -2.30
135 69.1 90 100 30.90 10.00 0.00 3.43 2.30 -
140 70.8 91.2 - 29.20 8.80 - 3.37 2.17 -
145 72.6 92.2 - 27.40 7.80 - 3.31 2.05 -
150 74 93.2 - 26.00 6.80 - 3.26 1.92 -

Page | 16
 155 75.3 93.9 - 24.70 6.10 - 3.21 1.81 -
160 76.6 94.6 - 23.40 5.40 - 3.15 1.69 -
165 78 95.2 - 22.00 4.80 - 3.09 1.57 -
170 79.2 95.9 - 20.80 4.10 - 3.03 1.41 -
175 80.3 96.5 - 19.70 3.50 - 2.98 1.25 -
180 81.3 96.9 - 18.70 3.10 - 2.93 1.13 -
185 82.5 97.4 - 17.50 2.60 - 2.86 0.96 -
190 83.3 97.8 - 16.70 2.20 - 2.82 0.79 -
195 84 98.2 - 16.00 1.80 - 2.77 0.59 -
200 84.8 98.6 - 15.20 1.40 - 2.72 0.34 -
205 85.7 98.9 - 14.30 1.10 - 2.66 0.10 -
210 86.2 99.2 - 13.80 0.80 - 2.62 -0.22 -
215 87 99.4 - 13.00 0.60 - 2.56 -0.51 -
220 87.5 99.7 - 12.50 0.30 - 2.53 -1.20 -
225 88.1 99.8 - 11.90 0.20 - 2.48 -1.61 -
230 88.6 100 - 11.40 0.00 - 2.43 - -

Agitation Rate (RPM) : Ln Oxygen Concentration versus Time

6.00
Ln Oxygen Concentration (C*-CL)(%)

5.00
4.00
3.00
2.00
1.00 y = -0.0078x + 5.2073

0.00
y = 0-0.0341x + 5.2645
-1.00 100 200 300 400 + 5.2885
y = -0.0117x 500 600 700 800
y = -0.0241x + 5.2296
-2.00
-3.00 y = -0.0294x + 5.1299
Time, t (s)

100RPM 200RPM 300RPM 400RPM

500RPM Linear (100RPM) Linear (200RPM) Linear (300RPM)
Linear (400RPM) Linear (500RPM)

Figure 5: Effect of Manipulated Agitation rate at Constant Aeration and Temperature

Figure 5 shows all the Ln oxygen concentration curve decreases over time. The higher the
agitation magnitude, the higher the rate of oxygen mass transfer as the value of the
volumetric mass transfer coefficient also increases. Table 5 shows the volumetric mass
transfer coefficient at varying agitation magnitude.

Page | 17
Table 5: Volumetric Mass Transfer Coefficient at Manipulated Agitation Rate

Agitation Rate 100 200 300 400 500

(RPM)
Volumetric Mass
Transfer 0.0078 0.0117 0.0241 0.0341 0.0294
Coefficient,
KLA(S-1)

Aeration Rate (L/min) : Ln Oxygen Concentration versus

Time
6.00
Ln Oxygen Concentration (C* - CL)(%)

5.00
4.00
3.00
2.00
1.00
y = -0.0241x + 5.2296
y = -0.0086x + 4.582
0.00
0 100 200 300 400 500 600
-1.00y = -0.0472x + 5.231
-2.00
-3.00
Time, t (s)

1 L/min 2 L/min 2.5 L/min

Linear (1 L/min) Linear (2 L/min) Linear (2.5 L/min)

Figure 6: Effect of Manipulated Aeration rate at Constant Agitation and Temperature

Figure 6 shows all the Ln oxygen concentration curve decreases over time. The higher the
aeration magnitude, the higher the rate of oxygen mass transfer as the value of the volumetric
mass transfer coefficient also increases. Table 6 shows the volumetric mass transfer
coefficient at varying aeration magnitude.

Table 6: Volumetric Mass Transfer Coefficient at Manipulated Aeration Rate

Aeration Rate (L/min) 1 2 2.5

Volumetric Mass
Transfer Coefficient, 0.0086 0.0241 0.0472
KLA(S-1)

Page | 18
 Temperature : Ln Oxygen Concentration versus Time
6.00
Ln Oxygen Concentration (C*-CL)(%)

5.00
4.00
3.00
2.00 y = -0.0103x + 4.8103

1.00
y = -0.0472x + 5.231 y = -0.0241x + 5.2296
0.00
0 50 100 150 200 250
-1.00
-2.00
-3.00
Time, t (s)

30C 35C 40C Linear (30C) Linear (35C) Linear (40C)

Figure 7: Effect of Manipulated Temperature at Constant Agitation and Aeration rate

Figure 7 shows all the Ln oxygen concentration curve decreases over time. The higher the
temperature, the higher the rate of oxygen mass transfer as the value of the volumetric mass
transfer coefficient also increases. Table 7 shows the volumetric mass transfer coefficient at
varying temperature.

Table 7: Volumetric Mass Transfer Coefficient at Manipulated Temperature

Temperature (°C) 30 35 40
Volumetric Mass
Transfer Coefficient, 0.0103 0.0241 0.0472
KLA(S-1)

Page | 19
 Volumetric Mass Transfer Coefficient against Agitation
Rate
0.04

0.035
Volumetric Mass Transfer

0.03
Coefficient, KLA(S-1)

0.025

0.02

0.015

0.01

0.005

0
0 100 200 300 400 500 600
Agitation Rate (RPM)

Figure 8: Volumetric Mass Transfer Coefficient against Agitation Rate

Volumetric Mass Transfer Coefficient against Aeration

Rate
0.05
0.045
Volumetric Mass Transfer

0.04
Coefficient, KLA(S-1

0.035
0.03
0.025
0.02
0.015
0.01
0.005
0
0 0.5 1 1.5 2 2.5 3
Aeration Rate (L/min)

Figure 9: Volumetric Mass Transfer Coefficient against Aeration Rate

Page | 20
 Volumetric Mass Transfer Coefficient against
Temperature
0.05
0.045
Volumetric Mass Transfer

0.04
Coefficient, KLA(S-1

0.035
0.03
0.025
0.02
0.015
0.01
0.005
0
25 27 29 31 33 35 37 39 41
Temperature (°C)

Figure 10: Volumetric Mass Transfer Coefficient against Temperature

Figure above shows the volumetric mass transfer coefficient at varying operating parameters:
(8) Agitation, (9) Aeration, and (10) Temperature. All the curves show increasing patterns
along the different magnitude of the respective operating parameters.

Page | 21
8.0 CALCULATION

Sample Calculation for Experimental Driving Force (Oxygen Concentration)

Driving Force = % DO at 100% Saturation - % DO at respective time
= C* - CL
= 100% - 0%
= 100%
Sample Calculation for Experiment Ln Driving Force (Ln Oxygen Concentration)
Ln Driving Force = Ln (C* - CL)
= Ln(100)
= 4.61

Sample Calculation for Slope of the Tangent of the Ln Oxygen Concentration Curve
𝐲 = 𝐦𝐱 + 𝐂
The linear equation is obtained by introducing the linear forecasting trendline to the plotted
graph of ln (C* - CL) against time.
y = -0.0078x + 5.2073
The slope of the ln oxygen concentration curve at agitation rate = 100 rpm, aeration = 2
L/min, and temperature = 30°C is m = – 0.0078.
Using the static gassing out method.

Sample Calculation for Volumetric Mass Transfer Coefficient, KLA

m = -KLA
KLA = -m
= -(-0.0078)
= 0.0078s-1
Using the static gassing out method.

Page | 22
9.0 DISCUSSION

The experiment is performed to investigate the effect of bioreactor parameters

[agitation rate (RPM), the aeration rate (L/min) and the temperature(°C)] on the
dissolved oxygen concentration, CL (%). This experiment is performed in the absence
of cell or bacterias, thus oxygen is not consumed throughout the experiment using the
static gassing out method. The data is taken until the value of the partial pressure of
oxygen gas reached to a constant value of approximately 100%. The mass transfer
coefficient signifies the rate of oxygen transfer between the gaseous and reaction liquid
(distilled water).

From the slope of the tangent to the curve of logarithm of driving force against time,
the mass transfer coefficient is attained. The driving force is also known as the dissolved
oxygen concentration. The partial pressure of the oxygen gas inside the reactor is
assumed to be the concentration of the dissolved oxygen inside the reactor. The system
is believed to reach maximum saturation point for oxygen gas as the partial pressure of
oxygen gas reached to constant.

The first parameter is the agitation. Aeration is a the movement of one or more
component of a mixture to improve contact (Dictionary, 2018). The contact between
the air and distilled water is improve as the agitation is increased. Increasing agitation
will increases the volumetric mass transfer coefficient. The volumetric mass transfer
coefficient at agitation rate equals to 100RPM, 200RPM, 300RPM, 400RPM and
500RPM are 0.0078s-1, 0.0117s-1, 0.0241s-1, 0.0341s-1 and 0.0294s-1, respectively.

From Figure 8, the greater the agitation magnitude, the greater the value of the mass
transfer coefficient. This also means that higher agitation will increase the rate of
oxygen transfer from air to the water. This is due to increasing in agitation will ensure
the well mixed of oxygen and water, since the speed of the stirrer is increases. The
movement of the stirrer, cutting the air and water in rapidly, increases the surface
contact area of both materials. Thus, increasing the rate of bioreactions However, at
500RPM, the volumetric mass transfer coefficient is going low. This is due to some
errors that occurs during the experiment.

Page | 23
The second operating parameter is the aeration rate. Aeration process of circulating air
through, mixed with or dissolved in a substance or liquid. In the experiment, this
parameter brings the water and air to close contact to removed dissolved gaseous. The
volumetric mass transfer coefficient at aeration rate equals to 1L/min. 2L/min and
2.5L/min are 0.0086s-1, 0.0241s-1, and 0.0472s-1, respectively.

From Figure 9. the higher the aeration magnitude, the greater the value of the
volumetric mass transfer coefficient. This also means that the rate of mass transfer of
oxygen from air to distilled water is increasing with increasing aeration since the air
collided more often with the water. The air can wonder throughout the reactor at greater
extend.

The third parameter is the temperature of the water. Temperature is the degree of
hotness of an object or substance with reference to some standard value. The volumetric
mass transfer coefficient at temperature equals to 30°C, 35°C and 40°C are 0.0103s-1,
0.0241s-1, and 0.0472s-1, respectively.

From Figure 10, the higher the temperature, the higher the magnitude of the volumetric
mass transfer coefficient. However, the increases in temperature only slightly increase
the value of the mass transfer coefficient. Based solely from Figure 10, temperature
does not show significant role in enhancing the rate of mass transfer in bioreactions.
Bioreactions usually involves enzymes in the reaction. However, increasing the
temperature could denature or damage the enzyme, thus high temperature is not suitable
for most bioreaction.

Based on all three parameters, the most substantial one is the agitation, followed by
aeration and temperature, respectively. Based on Figure 8, agitation shows higher
increment of the volumetric mass transfer coefficient compared to the other two
parameters. This concluded that, in the experiment, bioreactions is highly stimulated by
the increase of agitation. Furthermore, higher value or magnitude of the operating
parameters allow more oxygen to be in contact with the reaction liquid. This tell us that,
in the presence of bacteria or cell, more oxygen can be consumed, thus increasing the
growth rate of the bacteria

Page | 24
10.0 CONCLUSION

The method used for the MINIFORS (stirred tank bioreactor) is static gassing out
method. The volumetric mass transfer coefficient at agitation rate equals to 100RPM,
200RPM, 300RPM, 400RPM and 500RPM are 0.0078s-1, 0.0117s-1, 0.0241s-1, 0.0341s-
1
and 0.0294s-1, respectively. The greater the agitation magnitude, the greater the value
of the mass transfer coefficient. The volumetric mass transfer coefficient at aeration
rate equals to 1L/min. 2L/min and 2.5L/min are 0.0086s-1, 0.0241s-1, and 0.0472s-1,
respectively. The higher the aeration magnitude, the greater the value of the volumetric
mass transfer coefficient. The volumetric mass transfer coefficient at temperature
equals to 30°C, 35°C and 40°C are 0.0103s-1, 0.0241s-1, and 0.0472s-1, respectively.
The higher the temperature, the higher the magnitude of the volumetric mass transfer
coefficient. Based on three operating parameters, the most substantial one is the
agitation, followed by aeration and temperature, respectively. Since the experiment is
successfully done. Hence, objectives are successfully obtained.

Page | 25
11.0 RECOMMENDATION

To achieve a better result of experiment, some recommendations are suggested. Ensure

the air line or nitrogen line have no leakage before purging. Make sure the fixed
parameters are really constant before running the experiment by lets the reactor run for
a while. There are two method lies under the gassing out method to determine the mass
transfer coefficient, kLa which are dynamic and static. The student could use the
dynamic method instead of static method or even better used both and compared the
experimental value obtained from both methods. Another operating parameter can also
be consider using the MINIFORS which is pH of the reaction liquid. Basically, using
the same mechanism by using varying pH at constant another operating variable. Lastly,
the students should study the lab manual and prepare a necessary table before
conducting the experiment to ensure the experiment run smoothly.

Page | 26
12.0 REFERENCES

Fogler, H. S. (2006). Element of Chemical Reaction Engineering. Michigan: Prentice

Hall.
(2018). Lab Manual - Determination of The Volumetric Mass Transfer Coefficient in
Bioreactor.
Moutafchieva, D. (15 May,2013) Experiment Determination of the Volumetric Mass
Transfer Coefficient. Journal of Chemical Technology and Metallurgy, 48(4), 351-
356.
Organisms, M. (10 August, 2018). The Static Method of Gassing. Retrieved from
Buffalo
Brewing Blog: https://www.buffalobrewingstl.com/micro-organisms/the-static-
methodof-gassing-out.html
https://www.onlinebiologynotes.com/bacterial-growth-curve/

Page | 27
1.3 APPENDICES

Page | 28
Page | 29
Page | 30