FDE320 APPLIED KINETICS (19-20 Spring) 03.06.

2020
Instructor: Prof. Dr. Alev Bayındırlı
Assoc. Prof. Dr. Deniz Çekmecelioğlu

Assistants: Serap Namlı

Ülkü Ertuğrul

PROBLEM SET 12
1. A flat sheet of polymer coated with enzyme is placed in a stirred beaker. The maximum
rate of the enzyme is 6 x 10-6 mol/s-mg enzyme. The amount of enzyme bound to the
surface has been determined to be 1 x 10-4 mg enzyme/cm2 of support. In solution, a
value of Km is found to be 2 x 10-3 mol/l. The mass transfer coefficient was estimated
as 4.3 x 10-5 cm/s.

a) Determine the reaction rate when the bulk concentration is 7 x 10-3 mol/L.
(Ans: 4.66 10-6)

b) What is the controlling mechanism for this system? Why?

2. A wastewater stream of 1 m3/h has a low input concentration of a toxic compound (20
mg/L). The concentration must be reduced to 2 mg/L before discharge by using up-flow
packed bed containing surface immobilized enzyme in the form of beads.
Data:
V (mg/m3 bead-h) = Vmax S/Km+S = 0.2 S for [S]bulk<< Ks
a= 0.85 m3 bead/m3 of the bed
A bed = 4 m2
If [S]bulk<< Ks and the effectiveness factor=0.9, determine the height of the column
required. (Ans: 3.76 m)

3. The hydrolysis of sucrose at pH = 4.5 and 25°C using crude invertase obtained from
baker’s yeast in free and immobilized form was studied. The following initial velocity
data were obtained with 408 units of crude enzyme (1 unit = quantity of enzyme
hydrolyzing 1 μmol of sucrose/min when incubated with 0.29 M sucrose in a buffer at
pH 4.5 and 25°C).
FDE320 APPLIED KINETICS (19-20 Spring) 03.06.2020
Instructor: Prof. Dr. Alev Bayındırlı
Assoc. Prof. Dr. Deniz Çekmecelioğlu

Assistants: Serap Namlı

Ülkü Ertuğrul

V0 (mmol hydrolyzed/l-min)

Free Enzyme Immobilized Enzyme S0 (mol/l)

0.083 0.056 0.010

0.143 0.098 0.020

0.188 0.127 0.030

0.222 0.149 0.040

0.250 0.168 0.050

0.330 0.227 0.100

0.408 0.290 0.290

a. Determine the Km and Vmax for this reaction using both free and immobilized enzyme.
(Answer: Km = 0.05 M and Vmax = 0.5 mmol/l-min for free enzyme; Km = 0.05 M and
Vmax = 0.33 mmol/l-min for immobilized enzyme)

b. Do the data indicate any diffusion limitations in the immobilized enzyme preparation?