CALLUENG, JOMHEL B.

BSCHE-5

6.2 Aiba et al. (1968) reported the results of a chemostat study on the growth of a specific strain
of baker's yeast as shown in the following table. The inlet stream of the chemostat did not
contain any cells or products.

Inlet Concentration (g/L) Steady-state concentration (g/L)

Dilution Rate D, (hr-1 )
glucose CSi
Glucose CS Ethanol CP Cells CX
0.084 21.5 0.054 7.97 2.00
0.100 10.9 0.079 4.70 1.20
0.160 21.2 0.138 8.57 2.40
0.198 20.7 0.186 8.44 2.33
0.242 10.8 0.226 4.51 1.25

a. Find the rate equation for cell growth.

b. Find the rate equation for product (ethanol) formation

Solution:

a. Assume that the growth rate can be expressed by Monod kinetics and is represented by
 Cs
  max
Ks  Cs

For a chemostat, µ=D (Dilution rate),

 Cs
D  max
Ks  Cs

1 Ks  Cs

D  max Cs

1 Ks Cs
 
D  max Cs  max Cs
Plotting the graph of 1/D versus 1/Cs

1/Cs 1/D
18.51852 11.90476
12.65823 10
7.246377 6.25
5.376344 5.050505
4.424779 4.132231

Plot of 1/D vs 1/Cs for Baker's Yeast

14
y = 0.5585x + 2.081
12 R² = 0.9734
10

8
1/D

0
0 5 10 15 20
1/Cs

1/μmax 2.081 hr
μmax 0.480538 hr-1
Ks/μmax 0.5585 gL-1.hr
Ks 0.268381 gL-1

Substituting the values to the equation, we get

 Cs 
rx  Cx   max  Cx
 K s  Cs 

 0.480538Cs 
rx =μC x =   Cx
 0.2684+Cs 
 b. Assume that it is growth associated product we arrive at

1 dP
 YP / x 
Cx dt
dP
rP   YP / x  Cx
dt
dP  
rP   YP / x  max Cs  Cx
dt  Ks  Cs 
Where YP/x is the slope

Ethanol CP Cells CX
7.97 2.00
4.70 1.20
8.57 2.40
8.44 2.33
4.51 1.25

Plot of Cp vs Cx for Baker's Yeast

10
9
8
7
Ethanol Cp

y = 3.499x + 0.4139
6 R² = 0.9721
5
4
3
2
1
0
1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6
Cells Cx

The slope from the graph is YP/x=3.499. Substituting,

 0.480Cs 
rP =3.499   Cx
 0.2684+Cs 
6.7 Herbert et al.(1956) reported that the growth kinetics of Aerobacter cloacae in a chemically
defined medium (glycerol as a limiting substrate) could be expressed by Monod kinetics as
follows:

dCx maxCS C X
rx  
dt Ks  Cs

where µmax=0.85 hr-1 and Ks =1.23 x 10-2 g/L. The yield was found to be 0.53 g dry weight of
organism/g glycerol used. You are a biochemical engineer who has been assigned the task of
designing the most effective continuous fermentation system to grow the microorganism
(Aerobacter cloacae] with glycerol as its limiting substrate. For the following three questions,
the concentration of glycerol in the feed stream and that of glycerol in the outlet stream should
be 3 g/L and 0.1 g/L, respectively.

a. Since you have learned that the l/rx versus Cs curve for Monod kinetics has a U shape, you
have recommended that the most effective system would be the combination of a
continuous stirred-tank fermenter (CSTF) and a plug-flow fermenter (PFF). You were quite
sure of this because the substrate concentration in the outlet stream has to be so low.
However, your boss is insisting that the use of second PFF in addition to the first CSTF
will not improve the productivity very much. Who is right? Prove whether you are right or
wrong by drawing the 1/rx versus Cs curve for this microorganism. Does it have a U shape?
Discuss why you are right or wrong. (If you are right, think about how you can nicely
correct you boss's wrong idea. If you are wrong, it will teach you that you have to be careful
not to make a quick conclusion without adequate analysis.)
b. Recommend the best fermenter system (fermenter type and volume) which can handle 100
L/hr of feed stream. The best fermenter system is defined as that which can produce the
maximum amount of cells per unit time and volume.
c. If Ks = 1.23 g/L instead of 1.23 x 10-2 g/L, what is the best fermenter system (fermenter
type, volume) which can handle 100 L/hr in the feed stream. Draw the block diagram of
the fermenter system with the concentrations of the substrate and the cells in the inlet and
outlet streams of each fermenter. How is this case different from the case of part (a) and
why?
6.8 Suppose you have an organism that obeys the Monod equation:

dCx max CS C X

dt Ks  Cs

Where µmax=0.5 hr-1 and Ks=1 g/L.

The organism is being cultivated in a steady-state CSTF, where F = 100 L/hr, CSi = 50 g/L,
and YXIS = 0.5.

a. What size vessel will give the maximum total rate of cell production?
b. What are the substrate and cell concentrations of the optimum fermenter in part (a)?
c. If the exiting flow from the fermenter in part (a) is fed to a second fermenter (CSTF), what
should be the size of the second fermenter to reduce the substrate concentration to 1 g/L?
d. If the exiting flow from the first fermenter in part (a) is fed to a second fermenter whose
size is the same as the first, what will be the cell and substrate concentrations leaving the
second fermenter?

Given: max  0.5 hr 1

Ks  1 g / L

F  100 L / hr

Csi  50 g / L

YXIS  0.5

Solution:

a. Volume of vessel for maximum rate of cell production

Ks  Csi 1  50
 
Ks 1
  7.141
Csi 50 g / L
Cs  Csopt  
  1 7.141  1
Cs  6.141 g / L
  Cs  0.5 / hr  6.141g / L 
D  max 
Ks  Cs 1g / L    6.141g / L 
D  0.43 / hr

F F
D ;V 
V D
F 100 L / hr
V 
D 0.43 / hr
V=232.57 L

b. Substrate and cell concentrations

Csi 50
Cs  Copt  
  1 7.141  1
Cs=6.141 g/L
c.
6.9 You are going to cultivate yeast, Saccharomyces cerevisiae, by using a 10 m -fermenter your
company already owns. You want to find out the amount of ethanol the fermenter can produce.
Therefore, a chemostat study was carried out and the Monod kinetic parameters for the
microorganism grown in the glucose medium at 30°C, pH 4.8, were found to be: Ks = 0.0025
g/L and µmax = 0.25 h-1. The ethanol yield (YPIS) is 0.44 (gl g) and cell yield (YXIS) is 0.019
(g/g). The inlet substrate concentration is 50 g/L.

a. What flow rate will give the maximum total ethanol production in the continuous fermenter
and what is the maximum ethanol production rate?
b. If you want to convert 95 percent of the incoming substrate, what must the ethanol
production rate be for the continuous fermenter?
c. If you have two 5 m3 -fermenters instead of one 10 m3 -fermenter, what is your
recommendation for the use of these fermenters to convert 95 percent of the incoming
substrate? Would you recommend connecting two fermenters in series to improve the
productivity? Why or why not?

Given: max  0.25 hr 1

Ks  0.0025 g / L

YX / S  0.019 g / g

YP / S  0.44 g / g

Cso  50 g / L

Solution:

a. Flowrate for maximum ethanol production and maximum ethanol production rate

Ks  Csi 0.0025  50
 
Ks 0.0025
  141.425
Csi 50 g / L
Cs  Csopt  
  1 141.425  1
Cs  0.351 g / L
 
D  max 
Cs  0.25 / hr  0.351g / L 
Ks  Cs  0.0025 g / L    0.351g / L 
D  0.248 / hr
F
D ; F  DV
V
F  DV


F   0.248 / hr  10m3 
F=2.48m 3 /hr

b. Ethanol Production rate for 95% conversion

Cs  Cso 1  x 
Cs  50 1  0.95 
Cs  2.5 g / L


D  max 
Cs  0.25 / hr  2.5 g / L 
Ks  Cs  0.0025 g / L    2.5 g / L 
D  0.2498 / hr
F  DV


F   0.2498 / hr  10m3 
F=2.498m 3 /hr
c. Two 5-m3 fermenter or one 10-m3 fermenter
 YX / S   
Cxo  Cx1  YX / S Cso  
Cso  Cs   1 
0.019 g / g  141.425 
Cxo  Cx1  0.019  50   
50 g / L  2.5 g / L  141.425  1 
Cxo  4 x104 g / L Cx1  0.9433g / L

 Cso 
Cs1   
  1 

D1  max 
Cs  0.25 / hr  0.3511g / L 
 50 
Cs1    Ks  Cs  0.0025 g / L    0.3511g / L 
 141.425  1 
D1  0.248 / hr
Cs1  0.3511g / L

  
Cx2  YX / S Cs1  
F1  V1D1   1 

 
F1  0.248hr 1 5m3
 141.425 
Cx2  0.019  0.3511  
 141.425  1 
F1  1.2411m3 / hr Cx2  6.624 x103 g / L

Cx2  Cx1
 Cs  YX / S 
Cs2   1  Cs2  Cs1
  1 
 0.3511  6.624 x103  0.9433
Cs2    YX / S 
 141.425  1  2.465 x103  0.3511
Cs2  2.465 x103 g / L Y X/S =2.6875

Since the growth yield constant is higher than the 10m3 fermenter tank, a series of two 5-
m3 will be better.
6.10 You are a biochemical engineer in a pharmaceutical company. Your company is a major
producer of penicillin. Currently, what kind of fermenter is your company using for penicillin
production? Why? Your boss asked you to study the possibility of using an air-lift fermenter
as a replacement since it has many advantages. What is your recommendation?

Solution:

In most penicillin production companies, the antibiotic production requires a batch fermenter,
and a fed batch process is normally used to prolong the stationary period and so increase
production. On the other hand, the advantages on an air-lift fermenter are as follows

Advantages of an Airlift Fermenter

 Airlift fermenter is without any mechanical stirring arrangements

 Turbulence caused by fluid flow ensures adequate mixing of the liquid
 Compared to stirred tank airlift’s are easy to scale up & they require less energy to operate
 It is ideally suited for aerobic cultures since oxygen mass transfer coefficient are quite high.

One application of air-lift fermenters is the large scale production of monoclonal antibodies.
Considering this, it may an advantageous for the company to change into a newer technology
which offers many advantages. Even if the capital cost of obtaining one is high, the company will
surely benefit in the long run.