U

AGITATED AEROBIC FERMENTATION

Introduction
The power requirements for agitation and mixing in stirred gas-liquid reactors must be predicted
accurately in order to assure adequate gas-liquid mass transfer rates in support of the chemical
reaction(s) being carried out. Examples of commercial systems include chlorination of
polyisoprene (used in traffic paint), oxidation of p-xylene to make terephthalic acid,
hydrogenation of fats and oils to produce margarine, and aerated fermentation to make various
antibiotics. The analysis of a particular mixed reactor is often done at the lab or pilot plant scale,
and then results are scaled up using correlations based on dimensionless groups.
In this lab experiment, your company is moving into the production of alcohol by glucose
fermentation in semi-batch reactors. It has become necessary to specify to vendors the size of
the reactor vessels, the diameter and design of the impellers, the design of the baffles, and the
power and speed of the stirrer motors. Your group has been assigned to set up a pilot plant and
gather data on which these specifications will be based.
The key adjustable parameters are stirrer speed (N), impeller type (pitched or flat blade turbine),
air feed rate (Q), and pH. Necessary measured responses include dissolved oxygen
concentration (CL) and gassed (Pg) and ungassed (Po) power consumption. Other important
parameters include tank size with and without baffles and broth density and viscosity.
Continuous data acquisition allows the volumetric oxygen mass transfer coefficient (k La) to be
accurately determined so that the rate of oxygen mass transfer to the broth can be reliably
predicted.

Objectives
1. Obtain a correlation for the gas-liquid mass transfer coefficient (kLa) in aerated agitated
fermentors as a function of gassed power consumption (Pg), tank diameter (D), broth
volume (V), impeller diameter (Di), stirring rate (RPS), and superficial air velocity (Us) at
a fixed pH. Determine how closely the correlation fits the data obtained.
2. Obtain a correlation of the gassed power consumption as a function of the ungassed
power draw, impeller diameter and speed, and air feed rate.
3. Based on the laboratory results, design a 1000 L stirred fermentor to convert glucose to
alcohol, specifying tank diameter and height, impeller diameter and speed, gassed power
requirement, and air feed rate.

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

Theory
In fermentation processes involving the cultivation of aerobic microorganisms, oxygen is an
essential element in the supply of energy for cell metabolism and, therefore, a key element in the
synthesis of biomass and products. In an aerated stirred reactor, an impeller is used to disperse
gas into the liquid while providing enough mixing to achieve a homogeneous uniform
concentration of nutrients in the broth. Normally, the resistance to gas-liquid mass transfer
occurs primarily in the liquid film adjacent to the gas-liquid interface. A simple relationship can
be written that relates the gas mass transfer rate to the interfacial area, liquid film resistance, and
the concentration driving force:
M = kLa (C* CL)

(1)

where: M is the gas mass transfer rate (mg/L-s)

kLa is the overall mass transfer coefficient (s-1)
C* is the concentration of dissolved oxygen in the liquid at saturation (mg O2/L)
CL is the concentration of dissolved oxygen in the liquid (mg O2/L)
The overall mass transfer coefficient may be correlated with agitator power and gas feed rate as
follows:
kLa = A1 (Pg/V) (Us)

(2)

where: A1 is an empirical constant

and are dimensionless empirical constants
Pg is the power draw in a gassed system (kW)
V is the liquid volume (m3)
Us is the superficial gas velocity (m/s)
Values reported in the literature for the constants A1, , and vary considerably with the
geometry of the system, the range of variables covered, and the experimental method used.
Therefore, specific correlations for kLa to be used in this work should be obtained from benchscale experiments.
The gassed power consumption (Pg) can be correlated to the systems geometric parameters,
operating variables, and broth physical properties that are implicit in the ungassed power
consumption (Po). The power requirements in gassed and ungassed systems can be represented
by an equation developed by Michel and Miller:
Pg = A2 [ Po2 N Di3 / Q0.56]
where:
A2, and are empirical constants
Po is the power draw in an ungassed system (kW)
N is impeller speed (s-1)
Di is the impeller diameter (m)
Q is the air feed rate (SLPS)

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

(3)

The ungassed power consumption, in turn, can be estimated using the procedure outlined in
McCabe, Smith, and Harriott. [4] It has been shown that the power number (NP) is a function of
the Reynolds number (NRe), agitator type, and baffle design (Fig. 9-12) where
NRe = Di2 N /
(4)
and
NP = Po gc /(N3 Di5 )
(5)
Here, = liquid viscosity and = liquid density.
The volumetric oxygen mass transfer coefficient can be determined using the method of
Mignone and Ertola [5], based on a step change in stirrer speed during cultivation and outlined
by Badino and colleagues[6]. Assuming perfect mixing, the mass balance for liquid phase
dissolved oxygen during aerobic batch cultivation is
dCL = kLa (C* - CL) QO2 Cx
dt
where the first term on the right is the O2 transfer rate and the second term is the O2 reaction rate
(mg O2/Ls)(QO2 is the specific O2 uptake rate and Cx is the concentration of yeast). Using a
constant air feed rate (Q) and stirrer speed (N1), the culture reaches a pseudo-steady state I.
During this period, the mass transfer coefficient is (kLa)1 and the dissolved O2 is (CL1).
Increasing the stirrer speed to N2, a transient period (II) is produced, during which the dissolved
O2 concentration increases until a new pseudo-steady-state period (III) is reached. This new
period is characterized by a different mass transfer coefficient, (kLa)2, and a different dissolved
oxygen concentration (CL2). These periods can be described by the following equations:
Period I (Steady State)
dCL1 = (kLa)1 (C* - CL1) QO2 Cx = 0
dt

(6)

dCL = (kLa)2 (C* - CL) QO2 Cx

dt

(7)

dCL2 = (kLa)2 (C* - CL2) QO2 Cx = 0

dt

(8)

Period II (Transient Phase)

Period III (New Steady State)

For short periods, the reaction rate (QO2 Cx) can be assumed to be constant. Hence, from Eq.(8),
(kLa)2 (C* - CL2) = QO2 Cx
(9)
Substituting into Equation 9 into Equation 7,
dCL = (kLa)2 (CL2 - CL)
dt
with the initial condition:
t = 0, CL = CL1
Integrating,
CL = CL2 (CL2 CL1) exp(-(kLa)2 t)

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

(10)

(11)

Defining a dimensionless concentration,

= (CL - CL1)/(CL2 CL1)
Equation 11 can be written in dimensionless form:
= 1 - exp(-(kLa)2 t)

(12)

(13)

Equation 13 describes the liquid phase kinetics; however, it does not include the electrode
kinetics and the kinetics of the stagnant film that covers the electrode membrane. These can be
approximated by
d F = - F
(14)
dt
F
and
d E = F E
(15)
dt
E
where F is the dimensionless diffusion film response, E is the dimensionless electrode
response, F is the response time for the liquid film resistance, and E is the response time for the
electrode diffusion resistance.
CE (not dimensionless) is the concentration of dissolved oxygen reported by the electrode.
Combining and integrating Eqs. (13), (14), and (15) shows that if values of (1 - E) are plotted
versus time, the area (1) under the curve is given by

1 = (1 - E)dt = 1/(kLa)2 + E + F
0

(16)

The response times E and F are determined by a step change in oxygen concentration in the
medium in the absence of gas-liquid mass transfer effects, which can be produced by
instantaneously moving the electrode between two media with different oxygen concentrations.
The area (E) below the response curve versus time is then
E = (1 - E)dt = E + F

(17)

E is constant for a certain electrode. If we assume that the change in F does not change
appreciable with changes in stir speeds, we can use the E calculated from data collected at a
single stir speed (350 RPM).
Subtracting Equation17 from Equation16 yields
1 - E = 1/(kLa)2

(18)

By using this procedure, the mass transfer coefficient can be evaluated by a step change in stirrer
speed. Note that by returning the stirrer speed to its original value N1, the value of (kLa)1 can be
obtained using the same procedure.

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

Materials
Microorganism
Commercial Bakers yeast (Saccharomyces cerevisiae)
Culture Media
To activate and ferment the inocula, the culture medium will contain [g/L]:
Glucose (15.0)
KH2PO4 (5.0)
MgSO47H2O (0.5)
Yeast extract (3.0)
(NH4)2SO4 (4.5)
antifoam (1.0 mL/L)
The initial pH must be adjusted to and maintained at 4.6 using 4M HCl and 1M NaOH.
Apparatus and Procedure
The mixed reactor apparatus consists of the following major components:
12 L cylindrical reactor with heater
2 turbine blade agitators with 250W DC motor
Model 240 temperature control system
Neofox Oxygen Sensor
Model 260 pH electrode
Omega FMA3200 Mass Flow Controller
Condenser
Power indicator
Data logging software
Acid and base pumps
The experiments are to be carried out with 10 L of culture medium at 30oC and 4.6 pH using two
air feed rates and three stirring speeds. Data to be recorded include power requirements and
dissolved oxygen concentrations. The initial pieces of data to be taken will be the ungassed
power requirement (Po) at three stirring speeds. The power reported by the power meter is the
power going into the controller box, which is distributed into: 1) base power to run the controller
box itself, 2) power for the heater if it is operating, and 3) power for the impeller. We are
interested only in the last component (power for impeller), so you should figure out how to
account for the other two components.
The next task is to introduce air at 3 SLPM and determine the response curves to changes in
stirring rate at constant air sparge rate. When the stirring rate is changed, there will be a new
equilibrium dissolved oxygen concentration, and the response time to reach the new equilibrium
concentration is to be determined. Integration of the concentration-time data will yield a value
for 1. This should be done for the six combinations of air feed rate and stirring rate, starting at 3
SLPM and the lowest stirring rate and moving up to the next level. After equilibrium is reestablished, the stirring rate is moved to the third level. At each condition, the gassed power
requirement (Pg) should be recorded. The same steps are then run moving sequentially back
down to the lowest stirring rate. This is illustrated graphically on page 7. The entire procedure
is then repeated at the second sparge rate (6 SLPM).
ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

For each air feed rate, integration of the areas under the four response curves should yield three
values for 1. Values for kLa at each condition can then be calculated from Eq.(18). The value
for 1E can be determined from electrode response data that will be provided by the TA. In
addition, the empirical constants for Eq.(3) and Eq.(2) can be calculated by multiple regression.
Stepwise Procedure
1. The culture medium (10L) will be prepared by the TA and charged to a clean reactor and
inoculated 3 hours before the experiment. The airfeed will be set at 3 SLPM and the
stirrer at around 350 RPM. Note the oxygen concentration reading to see if the steady
state value is reasonable. If it is not, change the speed to a different value.
2. Set the logging mode for rapid.
3. Record the power used by the impeller.
4. Change the impeller speed in different steps similar to shown on page 7, e.g., to 450, then
500, then 450, and finally 350 RPM. At each RPM, note the power and wait for the
oxygen concentration to stabilize before changing speed.
5. Change the airfeed to 6 SLPM and repeat step 4.
6. Turn off the air feed and the agitator. Allow the liquid to degas. Turn the agitator back
on and determine the power draw for the 3 stirrer speeds.
Analysis of Results
1. Discuss what kLa indicates (why are there separate variables kL and a?). Calculate the kLas
for the six conditions in the experiment, taking into account the electrode kinetics. Discuss any
qualitative trends you notice between kLa, impeller speed, and sparge rate including possible
reasons for these trends.
2. Obtain a correlation for the mass transfer coefficient, k La, as a function of the gassed power
consumption, broth volume, and superficial gas velocity. Discuss the coefficients that you
obtained; are they reasonable in terms of their signs and magnitudes?
3. Obtain a correlation of the gassed power consumption as a function of the ungassed power
draw, impeller diameter and speed, and air feed rate. Is the power number given by Eq.(5)
constant? (Note: for Reynolds numbers > 1000, the NP should be fairly constant.)
4. Based on the laboratory results, you are to design a commercial-scale stirred fermenter
containing 1000 L of broth volume that transfers oxygen to the broth at the same rate per unit
volume as determined in the lab, specifying tank dimensions, air sparge rate, stirring rate, and
gassed power requirement. Consider how the aspect ratio of your scaled-up tank affects the
mass transfer rate and the power consumption. You should decide (with justification) what you
should/can try to optimize: power consumption, rate, air sparge rate, etc. Discuss the
uncertainties in your final design.

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

Typical Run History

N3
N2

N2
N1

ParameterValue

N1

CE3
CE2

CE2

CE1

CE1
Q

T1

T2

T3

T4

T5

Time

REFERENCES
1. Badino, A.C., Facciotti, M.C., and Schmidell, W., Volumetric Oxygen Transfer
Coefficients in Batch Cultivations Involving Non-Newtonian Broths, Biochem. Eng. J.
8, 111 (2001).
2. Cooper, C.M., Fernstrom, G.A., and Miller, S.A., Performance of Agitated Gas-Liquid
Reactors, Ind. Eng. Chem. 36(6), 504 (1944).
3. Michel, B.J. and Miller, S.A., Power Requirements of Gas-Liquid Agitated Systems,
AIChE J. 8 (2), 262 (1962).
4. McCabe, W.L., Smith, J.C., and Harriot, P., Unit Operations of Chemical Engineering,
5th ed., McGraw-Hill, New York (1993).
5. Mignone, C.F. and Ertola, R.J., Measurement of Oxygen Transfer Coefficient under
Growth Conditions by Dynamic Model Moment Analysis, J. Chem. Tech. Biotech. 34B,
121 (1984).
6. Badino, A.C., et al., Agitation and Aeration: an Automated Didactic Experiment,
Chem.Eng.Educ. 38(2), 100-107 (2004).

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)

Appendix I: Safety
1. Wear safety glasses and lab coats at all times.
2. Wear closed-toe shoes that give good traction on possibly wet floors.
3. Guard against electrical hazards by making sure that all equipment is well grounded using
three-wire plugs and other means. All instruments should be plugged into the power strip.
4. Think first of safety in any action you take. If not certain, ask the TA or a faculty member
before you act.
5. Loose clothing should not be worn near rotating equipment.
6. Impellers should be operating only when they are submerged no less than half way to the
bottom of the liquid in a mixing vessel.
Appendix II: Reactor Dimensions

Agitated Aerobic
Fermentor

7.125 in

15.375 in

0.625 in
15.75 in
6-blade
impellers

4.5 in

0.625 in

3.25 in

6.625 in

7.75 in

ChBE4200/4210 Agitated Aerobic Fermentation (revised 5/23/16)