Bioprocess Engineering

Block 3. ENZYMATIC REACTIONS

4. The enzyme E catalyzes the transformation of S into the product R in the
1. The following data were obtained during the study of the catalytic activity following way:
of an intestinal peptidase able to hydrolyze the dipeptide glycylglycine to 200.CS CE
produce two moles of glycine: S + E  R + E  rS  mol / l.min
2  CS
S (mM) 1.5 2.0 3.0 4.0 8.0 16.0
If enzyme (EC = 0.001 mol / l) and reagent (CS0 = 10 mol /l) is introduced in
v (mM/min) 0.21 0.24 0.28 0.33 0.40 0.45 a batch reactor, letting the reaction progress. Calculate the necessary time for
the reagent concentration falls to 0.025 mol /l.
Determine the vaules of KM y Vmax for this enzymatic preparation.

2. The kinetics of an enzymatic hydrolysis reaction corresponding to the 5. Experiments carried out with enzyme E demonstrate that from 20°C the
following reaction scheme S + E  P have been studied. Experiments enzyme undergoes a thermal denaturation. The time required to deactivate
have been carried out in a stirred tank reactor. The enzyme is free and 50% of the enzyme at different temperatures has been determined:
suspended in the solution. The results obtained are shown in the following
table:
T (ºC) 25 30 40 45
t1/2 (min) 44.15 30.0 7.78 3.96
v (mol S/l.h) [E]T (gE/l) [S] (mol S/l)
0.15 0.1 0.3 Plot the value of kd against the temperature and deduce an expression of
0.20 0.1 0.6 variation with temperature.
0.45 0.3 0.3

Determine the parameters of the kinetic equation, assuming a Michaelis-

Menten type model for this reaction.

3. Catalase has been used to accelerate the decomposition of hydrogen

peroxide (hydrogen peroxide) to water and oxygen. The evolution of the
concentration of hydrogen peroxide with time at pH = 8.76 and T = 30ºC,
is given in the following table:

a) Determine the parameters of that equation, considering that the kinetic

equation is of the Michaelis-Menten type.
b) If the concentration of enzyme used is tripled. What concentration of
substrate will remain without reacting after 20 min?