Questions for PhD qualifiers: Bioprocess Engineering

PROBLEMS FROM BIOPROCESS ENGINEERING SHULER AND

KARGI
CHAPTERS 3, 7, 9 and 10
Q1. The hydrolysis of urea by urease can be described by Michealis-Menten
kinetics with the maximum reaction rate Vmax = 5 moles/l-min and Km =
0.05 M. Determine the time required to consume 90% of the initial urea
concentration (S0), when (i) S0 = 10 moles and (ii) S0 = 0.1 mole?
Q2. Evaluate the pH value at which the maximum enzymatic rate occurs for
the below pH dependent mechanism for a given substrate concentration of S0
assuming Haldanes approach?

Q 3. Enzyme inhibition kinetics: The following data was obtained for an

enzyme E for a fixed substrate concentration S0
Substrate
concentration

Inhibitor
concentration

v/vmax

S0

(mM)

50

0.9

50

10

0.75

25
10
0.6
Determine the enzymatic rate as a fraction of vmax for a substrate
concentration of 10mM and an inhibitor concentration of 25 mM.
Q1. Lactobacillus bulgaricus is used to ferment milk to yogurt. The growth
rate of L. bulgaricus on lactose (S) in milk is given as follows:

0.4
P
1
10

where P is the lactic acid concentration. It is known that L. bulgaricus

produces lactic acid only while it is growing. A chemostat (CSTR) is
operated for the preproduction stage, where the input to the reactor is
sterilized milk with 5 g/L of lactose. The outlet stream contains 3 g/L of
Lactose. It is known that YX/S = 0.5 and YP/X = 2 for L. bulgaricus. Another
organism has contaminated the fermentation, whose growth rate is given by

0.32S
0.2 S

Will the organism spoil the chemostat operation through contamination?

Q2. Fed-Batch Operation: A fermentor is operated as a fed-batch process.
The substrate concentration is kept constant in the reactor at S. The feed
stream has a substrate concentration of Sf. The growth rate of the organism is
represented by Monods kinetics. Determine the feed flow rate for the
process and determine the final biomass concentration? Assume Yx/s as the
yield coefficient?
Q3. In a fed batch culture operating with intermittent addition of glucose
solution, values of the following parameters are given at time t=2 h, when
the system is at quasi-steady state. V =1000 ml, F=200ml/h, S0 = 100g/l,
mumax=0.3 1/hr, Ks=0.1g/l, Yx/s=0.5g/g, X0=50 g/l
Find V0 (initial volumn). Determine the concentration of biomass at t=2h
assuming quasi steady state. If alpha=0.2g/g Determine the concentration of
product at t=2h assuming P0=0.

Q4. In a chemostat with cell recycle, the feed flow rate and culture volumes
are F=100ml/h and V=1000ml, respectively. The system is operated under
glucose limitation and the yield coefficient, Yx/s =0.5g/g. Glucose
concentration in the feed is S0=10g. The kinetic constants of the organisms
are mumax=0.21/h, Ks=1g/l. The value of C is 1.5 and the recycle ratio is
0.7. The system is at steady state. Find the substrate concentration in the
recycle stream Find the specific growth rate of the organisms. Find the cell
concentration in the recycle stream. Find the cell concentration in the
centrifuge effluent.
Q2. A methanol utilizing strain has a specific growth rate given by:
0 .2

s
0 .5 s s

50

Methanol concentration in the feed solution is 100 g/L with an initial

cellmass of 20 g/L and an initial volume of reactor of 500L. Determine the
final volume and cellmass concentration for operating a fed batch reactor at
maximum growth rate in the reactor. The yield coefficient Y X/S = 0.4 g dw
cells/g glucose. [Marks: 3.5]
Q3. Determine the dilution rate, amount of substrate consumed, product and
biomass formed to maximize (a) product productivity and (b) Biomass
Productivity for an organism with the following growth parameters:
P

0.6 1

40

; growth related product formation constant, alpha = 2; non

growth associated constant, beta=1 and YX/S = 2 g/g [Marks 3.5]

Q2 Chemostat Operation: Lactobacillus bulgaricus is used to ferment milk
to yogurt. The growth rate of L. bulgaricus on lactose (S) in milk is given as
follows:

0.4
P
1
10

where P is the lactic acid concentration. It is known that L. bulgaricus

produces lactic acid only while it is growing. A chemostat (CSTR) is
operated for the preproduction stage, where the input to the reactor is
sterilized milk with 5 g/L of lactose. The outlet stream contains 3 g/L of

Lactose. It is known that YX/S = 0.5 and YP/X = 2 for L. bulgaricus. Another
organism has contaminated the fermentation, whose growth rate is given by

0.32S
0.2 S

Will the organism spoil the chemostat operation through contamination?

Justify your solution with an explanation.
Q1. Consider the following reaction sequence:
S + E (ES)1 (ES)2 P + E
Develop a suitable rate expression for the product formation (dP/dt =
k4(ES)2) assuming quasi-steady state approximation?