1. The Zymomonas mobilis cells are used for chemostat culture in a 60 m3 fermenter.

The feed
contains 12 g l-1 glucose; Ks for the organism is 0.2 g l-1. m= 0.3 h-1 and Yx/s= 0.06 gdw/g.
a. What flow rate is required for a steady-state substrate concentration of 1.5 g l-1 ?
b. What is the corresponding steady state cell density and volumetric cell production rate?

1. Consider a continuous, aerobic bacterial culture in a chemostat with sterile feed. Three
different dilution rates D are tested for a glucose feed concentration Sf = 10 mM, and the biomass
concentration x and glucose concentration S in the exit stream are measured. The results are as
follows:
D (h-1)
0.05
0.5
5

x (g/L)
0.248
0.208
0

S (mM)
0.067
1.667
10

a. Estimate the glucose yield coefficient YX/S (g biomass/mole glucose).

b. Assuming Monod growth kinetics, estimate the maximum specific growth rate max (h-1) and
the Monod constant KS (mM).
Ans.
i.

Between time t = 0.05 to 0.05

YX / S

C X 0.208 0.248 0.04

0.025
1.667 0.067 1.6
C S

Between time t = 0.5 to 5

YX / S

ii.

C X 0.0 0.208 0.208

0.025
10 1.667 8.333
C S

Monod chemostat model equation

maxs

D x Dx f 0
Ks s

For sterile feed xf = 0


DK s
DK s
and ssterile feed
xsterile feed YX / S s f
m ax D
max D

(any of this two equation could be use to solve the problem)

D=
At 0.05 h-1
0.067

0.05 K s
m ax 0.05

(1)

0.5 K S
MAX 0.5

(2)

At 0.5 h-1
1.667

Solve equation (1) and (2) to get MAX (= 0.802) and KS (=1.007)

3. Consider a culture of bacteria making a valuable product in a chemostat operated at steady

state. The liquid feed is sterile and contains 50 mM glucose (the limiting growth substrate S). In
a series of steady state runs, the dilution rate D is increased incrementally, and the cell density
exiting the chemostat is measured for each:
D (h-1)
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
0.50
0.52
0.53

x (g/L)
1.58
2.17
2.47
2.65
2.76
2.83
2.86
2.84
2.74
2.25
1.32
0

a. Explain why the cell density can increase as the flow rate is increased, as shown here with D <
0.35 h-1.
b. At what dilution rate should you operate the chemostat to optimize productivity of a strictly
growth-associated product? Explain the basis for your answer.
c. At what dilution rate should you operate the chemostat to optimize productivity of a strictly
non-growth-associated product? Explain the basis for your answer.
4. Consider cell growth in a chemostat at steady state, with sterile feed. In bench-scale
experiments, it is found that the specific growth rate is inhibited at high glucose concentrations S,
modeled according to the phenomenological expression
(h-1) = 0.6S/(0.2 + S + 0.1S2), with S in g/L.
a. If the glucose concentration in the feed is 10 g/L, estimate the minimum dilution rate at which
cell washout would occur.
b. Is the dilution rate estimated in part a the highest that will support cell growth in the
bioreactor? Justify and briefly explain your answer.
5. A new strain of yeast is being considered for biomass production. The following data were
obtained using a chemostat. An influent substrate concentration of 800 mg/l and an excess of
oxygen were used at a pH of 5.5 and T= 350C. Using the following data, calculate m, Ks, YX/SM,
kd and ms assuming net=mS/(Ks + S)- kd.
Dilution
rate (h-1)

0.1
0.2
0.3
0.4
0.5
0.6
0.7

Carbon
substrate
concentration
(mg/l)
16.7
33.5
59.4
101
169
298
702

Cell
concentration
(mg/l)
366
407
408
404
371
299
59

6. The specific growth rate for unlimited growth in a chemostat is given by the following
equation:

where
S0 = 10 g/l

Ks = 1 g/l

I= 0.05 g/l

YX/SM= 0.1 g cells/g subs

X0=0

KI=0.01 g/l

m= 0.5 h-1

kd=0

a. Determine X and S as a function of D when I=0.

b. With inhibitor added to a chemostat, determine the effluent substrate concentration and X
as a function of D.
c. Determine the cell productivity, DX, as a function of dilution rate.
7. Pseudomonas sp. has a mass doubling time of 2.4 h when grown on acetate. The saturation
constant using this substrate is 1.3g / l (which is unusually high), and cell yield on acetate is
0.46gcell / g acetate. If we operate a chemostat on a feed stream containing 38 g/l acetate, find
the following:
a.
b.
c.
d.

Cell concentration when the dilution rate is one-half of the maximum

Substrate concentration when the dilution rate is 0.8 Dmax
Maximum dilution rate
Cell productivity at 0.8 Dmax

8. Under substrate-limited conditions, a microorganism exhibits the following net specific

growth rate, net, and yield coefficient, YX/S:
net (h-1) = 0.70 S/(0.1 + S), with S in g/L;
YX/S (g DCW/g) = 0.40
The available growth medium contains 10 g/L substrate.
a. When a batch bioreactor containing 100 L of the growth medium is inoculated with 1.0 g
DCW of biomass, estimate the maximum cell density achieved, and the approximate time
required to achieve it, after exponential growth is initiated.
b. You decide instead to culture the microorganism in a chemostat, using the same growth
medium as the (sterile) feed. Estimate the dilution rate (h-1) at which the chemostat will achieve
maximum steady-state productivity of biomass.
c. Calculate and compare the overall biomass productivities (g DCW/L/h) of the two scenarios in
parts a & b. What other consideration will make the batch process even less productive
compared to the chemostat?

8. The following data are obtained in oxidation of pesticides present in wastewater by a mixed
culture of microorganisms in a continuously operating aeration tank.
S.No.

D ( h 1 )

S ( Pesticides), mg / l

X (mg / l )

1.

0.05

15

162

2.

0.11

25

210

3.

0.24

50

250

4.

0.39

100

235

5.

0.52

140

220

6.

0.7

180

205

7.

0.82

240

170

Assuming the pesticide concentration in the feed wastewater stream as S 0 500 mg/L,
determine Y XM/ S , k d , m , and K s .