5. Table 3 was obtained in a chemostat for the growth of E.

aerogenes on a glycerol limited

growth medium. For this system, estimate the values of:
a. KS, g-glycerol/L
b. μmax, h-1
c. YX/S, g-cells/g-glycerol
d. mS, g-glycerol/(g-cell.h)
Dilution rate D, h-1 Glycerol Concentration S, g/L Cell concentration X, g/L
0.05 0.012 3.2
0.10 0.028 3.7
0.20 0.05 4.0
0.40 0.10 4.4
0.60 0.15 4.75
0.70 0.176 4.9
0.80 0.80 4.5
0.84 10.00 0.5
NOTE: S0=10 g/L

Given:

Dilution rate D, h-1 Glycerol Concentration S, g/L Cell concentration X, g/L

0.05 0.012 3.2
0.10 0.028 3.7
0.20 0.05 4.0
0.40 0.10 4.4
0.60 0.15 4.75
0.70 0.176 4.9
0.80 0.80 4.5
0.84 10.00 0.5
Required:

a. KS, g-glycerol/L
b. μmax, h-1
c. YX/S, g-cells/g-glycerol
d. mS, g-glycerol/(g-cell.h)

Solution:
𝐴𝑃 𝐴𝑃
D S X ∆S 𝑌𝑋/𝑆 1/𝑌𝑋/𝑆 1/D ug 1/ug 1/S
0.05 0.012 3.2 9.988 0.320 3.121 20.000 0.078 12.820 83.333
0.10 0.028 3.7 9.972 0.371 2.695 10.000 0.128 7.813 35.714
0.20 0.05 4 9.950 0.402 2.487 5.000 0.228 4.386 20.000
0.40 0.10 4.4 9.900 0.444 2.250 2.500 0.428 2.336 10.000
0.60 0.15 4.75 9.850 0.482 2.074 1.667 0.628 1.592 6.667
0.70 0.176 4.9 9.824 0.499 2.005 1.429 0.728 1.374 5.682
 0.80 0.8 4.5 9.200 0.489 2.044 1.250 0.828 1.208 1.250
0.84 10 0.5 0.000 --- --- 1.190 0.868 1.152 0.100

Assuming that extracellular product formation is negligible and the system is at steady-state, we have:
1 1 𝑚𝑠
𝐴𝑃 = 𝑀 + 𝐷
𝑌𝑋/𝑆 𝑌𝑋/𝑆
1
Plotting 𝐴𝑃 vs. 1/D:
𝑌𝑋/𝑆

3.5

2.5 y = 0.0578x + 2.0368

R² = 0.9382
2
1/YX/S

1.5

0.5

0
0 5 10 15 20 25
1/D

Thus, 𝑚𝑠 :
𝑔 − 𝑔𝑙𝑦𝑐𝑒𝑟𝑜𝑙
𝑚𝑠 = 0.0578
𝑔 − 𝑐𝑒𝑙𝑙 ℎ𝑟

In determining KS and μmax, recall from the Monod equation:

1 𝐾𝑆 1
= +
𝜇 𝑆𝜇𝑚𝑎𝑥 𝜇𝑚𝑎𝑥
Plotting 1/ 𝜇 vs. 1/S:
 16

14 y = 0.1483x + 1.0687
R² = 0.9746
12

1/μ 10

0
0 10 20 30 40 50 60 70 80 90
1/S

Solving for μmax:

1
= 1.0687
𝜇𝑚𝑎𝑥

𝜇𝑚𝑎𝑥 = 0.937 ℎ−1

Solving for Ks:
𝐾𝑆
= 0.1483
𝜇𝑚𝑎𝑥
𝐾𝑠 = (0.1483)( 0.937)

𝐾𝑠 = (0.1483)( 0.937)

𝐾𝑠 = 0.139 𝑔 − 𝑔𝑙𝑦𝑐𝑒𝑟𝑜𝑙/𝐿

Solving for YX/S:

∆𝑋
𝑌𝑋/𝑆 =
∆𝑆
Assuming X0=0,
3.2
𝑌𝑋/𝑆 =
9.98
𝑔 𝑐𝑒𝑙𝑙
𝑌𝑋/𝑆 = 0.320
𝑔 𝑔𝑙𝑦𝑐𝑒𝑟𝑜𝑙
 6. Lactobacillus casi is propagated under essentially anaerobic conditions to provide a
starter culture for manufacture of Swiss cheese. The culture produces lactic acid as a by-
product of endogenous metabolism. The growth parameters are given by Y X/S = 0.23 g-
X/g-S, KS = 0.15 g/L, μmax = 0.33 h-1, kd = 0.03 h-1. A stirred tank fermentor is operated in
fed-batch mode at pseudo-steady state with a feed flow of 4000 L/h and feed substrate
concentration of 80 g/L. The feed is started such that pseudo-steady state is reached from
time zero. After 8 h, the liquid volume is 40,000 L.
a. What was the initial culture volume?
b. What is the concentration of cell biomass at pseudo-steady state?
c. What is the mass of cells produced after 8 h fed-batch operation?
d. What is the substrate concentration in the reactor after 8 h fed-batch operation?

Given:
YX/S = 0.23 g-X/g-S
KS = 0.15 g/L
μmax = 0.33 h-1
kd = 0.03 h-1
F= 4000 L/h

S= 80 g/L

V(8 hrs)=40,000 L

Required:

a. Initial Culture Volume (V0)

b. Concentration of cell biomass at pseudo steady-state
c. Mass of cells produced after 8 h fed batch operation,
d. Substrate concentration in the reactor after 8 hr, S(8 hr)

Solution:

a. To solve for the initial culture volume,

Doing a mass balance over the reactor
𝑑𝑚
= 𝑚𝑖𝑛̇
𝑑𝑡
𝑑(𝜌𝑉)
= 𝜌𝐹
𝑑𝑡
Assuming dilute solution, such that the density is constant,
𝑑𝑉
=𝐹
𝑑𝑡
 Integrating, we get,
𝑉 = 𝑉0 + 𝐹𝑡
Solving for 𝑉0 ,
𝑉0 = 𝑉 − 𝐹𝑡
𝐿
𝑉0 = 40,000 𝐿 − (4000 )(8 ℎ𝑟𝑠)
ℎ
𝐿
𝑉0 = 40,000 𝐿 − (4000 )(8 ℎ𝑟𝑠)
ℎ
𝑉0 = 8,000 𝐿

b. To solve for the cell concentration,

𝑋 = 𝑌𝑋/𝑆 (𝑆0 − 𝑆)
𝑔−𝑋 𝑔 𝑔
𝑋 = (0.23 )(80 − 0.065 )
𝑔−𝑆 𝐿 𝐿
𝑋 = 18.39 𝑔/𝐿

c. Solving for the mass of cells produced,

𝑚𝑎𝑠𝑠 𝑜𝑓 𝑐𝑒𝑙𝑙𝑠 = 𝑋(𝑉 − 𝑉0 )
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑐𝑒𝑙𝑙𝑠 = 18.39 𝑔/𝐿(40,000 𝐿 − 8,000 𝐿)
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑐𝑒𝑙𝑙𝑠 = 588, 480 𝑔 𝑐𝑒𝑙𝑙𝑠
d. Recall the Monod equation for Specific Growth rate and at pseudo steady-state 𝜇 = 𝐷,
𝜇𝑚𝑎𝑥 𝑆
𝜇=𝐷=
𝐾𝑠 + 𝑆
𝐷(𝐾𝑠 + 𝑆) = 𝜇𝑚𝑎𝑥 𝑆
𝑆(𝜇𝑚𝑎𝑥 − 𝐷) = 𝐷 𝐾𝑠
𝐷 𝐾𝑠
𝑆=
(𝜇𝑚𝑎𝑥 − 𝐷)
𝐿
4000 𝑔
(40,000ℎ𝐿) (0.15 𝐿 )

𝑆=
𝐿
4000
0.33 ℎ−1− ℎ
40,000 𝐿
𝑆 = 0.065 𝑔/𝐿