218

Practical Fermentation Technology

e
*g
gen
i
i
l
L
max
m
N
o
O
P
S
S
v
x
xd
xv

endogenous
in equilibrium with gas phase
generation
inlet
inhibition
lysis
liquid phase
maximum
maintenance
nitrogen
outlet
oxygen
product
substrate
saturation
viable
cells
non-viable cells
viable cells
(prime) superscript for yield factor

Appendix: Problems and Solutions

Batch Culture
What can You Do with the Experimental Data?
The experimental data obtained in a microbial batch culture are given in Table A.1. N
refers to the nitrogen source and S indicates the carbon and energy source.
Find the volumetric rate expressions of growth, substrate consumption and product
formation and estimate the numerical values of the relevant kinetic parameters. In doing
so,
(a) Estimate the values of the maximum specific growth rate, mmax and the Monod coefficient, KS from the LineweaverBurk plot.
(b) Estimate the values of the maintenance constant, mS and the yield for biomass on
1
.
carbon substrate,
Yx/S

1
(c) Estimate the value of the yield for biomass on nitrogen substrate,
.
Yx/N

Solution
First plot all the concentrations against time and try to plot smooth curves (or lines, if it
looks linear) through the experimental points either by hand or using a suitable graphics
programme such as Excel. At this stage we do not know the mathematical expression (or
the model) that would give us how the concentrations change with time. We shall use

Biological Activity in Fermentation Systems

219

Table A.1 Experimental data obtained in a microbial batch

culture
t (h)

xv (kg/m3)

S (kg/m3)

N (kg/m3)

0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

0.100
0.134
0.180
0.241
0.323
0.433
0.581
0.778
1.040
1.400
1.870
2.500
3.350
4.490
6.000
8.000
10.700
14.100
17.900
18.300
18.300

40.00
39.93
39.83
39.20
39.50
39.30
39.10
38.50
37.80
37.20
35.40
34.80
32.90
29.50
27.20
21.80
17.10
9.60
1.11
0.00
0.00

4.00
4.00
3.99
3.98
3.97
3.96
3.94
3.92
3.88
3.84
3.78
3.70
3.59
3.44
3.24
2.97
2.57
1.89
1.50
1.49
1.48

these smooth curves through the data points in the calculation of time derivatives,
d(. . .)/dt.
We assume that growth follows Monod kinetics and the growth limiting nutrient is the
carbon and energy source, S. Since there is no decrease in the values of the biomass
concentration at the end of the batch, we can also assume that microbial death can be
neglected.
(a) Estimation of max and KS from the LineweaverBurk plot. The LineweaverBurk
plot is given by Equation (7.76):
We need to plot 1/m versus 1/S and evaluate KS from the slope, and 1/mmax from the yaxis intercept. We therefore need to obtain the values of the specific growth rate m which
changes with the carbon substrate concentration S, which in turn changes with time, t.
From the mass balance on biomass in a batch culture, using Equations (7.57), (7.61) and
(7.62), and assuming that there is no death, we have:
dx v
= rx = x v
dt

so that

1
1 dx
= v
x v dt

According to this, 1/m value at a time t can be obtained from the time derivative of cell
concentration, dxy/dt. For this, use the plot of experimental cell concentration against time,
as shown in Figure A.1.

220

Practical Fermentation Technology

20

Method 1:

Dry weight, x v ( kg cells/m 3)

18

dxv 11.9
=
= 1.700
7
dt

16
14

Method 2:

12

dxv x15 x13 8.00 4.49

= 1.755
=
=
15 13
dt
t15 t13

10
8

11.9

6
4
7

2
0
0

12

16

20

Time (h)

Figure A.1 Biomass concentration against time plot and the slope-of-the-tangent method to
obtain values at different times

How to obtain the time derivatives, d(. . .)/dt:

Method 1: Hand-drawn tangent: Hand draw a tangent to the smooth curve drawn through
the experimental data points and measure its slope as shown in Figure A.1 for t = 14 h.
Method 2: Numerical differentiation: Take the difference between the values equally
spaced on either side of the data point being analysed (t = 14 h) and divide by the appropriate time interval as shown in Figure A.1.
Method 3: Curve fitting: You may have already used a software to plot a smooth curve
through your experimental data points. Some software gives you the equation used for
curve fitting and from the equation you can find the value of the time derivative.
Some values of 1/m obtained from the experimental data are given in Table A.2.
Use the data in Table A.2 to obtain the LineweaverBurk plot of Figure A.2
From the y-axis intercept on can calculate:
1
= 0.294 h 1
3.4
From the x-axis intercept, one can calculate:

max =

1
= 1.67 kg m 3 glucose.
0.6
KS
These values can be checked from the slope of the line,
= 5.67 kg glucose h m3 as
max
shown in Figure A.2.
KS =

Biological Activity in Fermentation Systems

221

Table A.2 Calculation of 1/ and 1/S for the LineweaverBurk plot

t (h)

xv (kg/m3)

rx = dxv/dt

m = rx/xv

1/m

1/S

0
1
2
3
...
12
13
14
15
16
17
18

0.100
0.134
0.180
0.241
...
3.350
4.490
6.000
8.000
10.700
14.100
17.900

0.040
0.054
0.072
...
0.995
1.325
1.755
2.350
3.050
3.600

0.299
0.297
0.297
...
0.297
0.295
0.293
0.294
0.285
0.255

3.350
3.364
3.371
...
3.367
3.389
3.419
3.404
3.508
3.917
8.547

40.00
39.93
39.83
39.70
...
32.90
30.50
27.20
22.80
17.10
9.60
1.11

0.0250
0.0250
0.0251
0.0252
...
0.0304
0.0328
0.0368
0.0439
0.0585
0.1042
0.9009

9.0
8.0
1/m (h)

7.0

Slope =

6.0

mm

5.0

=
3.0

1
= 0.6
KS

1
= 3.4h
mm

2.0
1.0

-0.60

8.5 3.4
0.9

4.0

KS

0.0
-0.10

0.40

= 5.67

kg glucose
hm3

0.90

1/S (m /kg glucose)

Figure A.2 LineweaverBurk plot

(b) Estimation of the maintenance constant mS and biomass yield on carbon source, Yx/S.
Inserting Equations (7.6) and (7.37) into Equation (7.50) (or from Equation (7.65) and
assuming that there is no product formation other than the cells, we have:
rS =

x
rx
+ mS x v = v + mS x v
Yx/S
Yx/S

Dividing both sides with xv we get:

1 rx
rS
=
+ mS
x v Yx/S x v