Tutorial 5
Tutorial 5
Tutorial 5
1 2
+ +
k-1
a. Calculate the ratio of the two effectiveness factors, (E0 = 20 M)/(E0 = 10 M).
b. Calculate the ratio of the two Thiele moduli, (E0 = 20 M)/(E0 = 10 M).
Solution to Tutorial 5
1. Known: reaction mechanism; rate constants; and enzyme and substrate concentration.
Analysis:
First, we noted that the reaction mechanism is the same mechanism assumed in the derivation
of the Michaelis-Menten equation. Therefore the product formation rate will be given by:
dP vmax [ S ] k2 [ E0 ][ S ]
= =
dt km + [ S ] km + [ S ]
The half-saturation constant, km, will then be a function of the individual rate constants as
shown in the derivation of the Michaelis-Menten equation:
k1
km = , if the rapid equilibrium assumption is used, or
k1
k1 + k2
km = , if the quasi-steady state assumption is used.
k1
Therefore:
k1 4.4 104 s 1
km = = 9 1 1 = 4.4 10 5 M [rapid equilibrium]
k1 10 M s
Where the value used for km was the one calculated using the quasi-steady state assumption.
2. Known: initial reaction rate data for free and immobilized enzymes after treatment at
80C for different lengths of time.
Analysis:
Let E0 be initial amount of enzyme before thermal treatment, and E0(t) the amount of
active enzyme after time t operating at 80C.
Recalling that the maximum reaction rate is given by (as per Michaelis-Menten
mechanism):
v max = k 2 [ E 0 ]
Assuming a first order degradation kinetics for the enzyme thermal degradation:
[ E0 (t )] = [ E0 ] e kd t
Then the maximum reaction rate at different times can be written as:
vmax (t ) = k 2 [ E0 (t )] = k 2 [ E0 ] e kd t = vmax e kd t
Recall the Michaelis-Menten equation:
v max [ S ] k 2 [ E 0 ][ S ]
v= =
k m + [S ] k m + [S ]
At a constant substrate concentration [S], the reaction rate is directly proportional to vmax,
and we can use v instead of vmax:
E (t ) ln 2
[ E0 (t )] = [ E0 ] e kd t ln 0 = k d t t1 / 2 =
E0 kd
t1/2 = 6.5 min (free soluble enzyme)
3. Known: Intrinsic kinetic parameters, effective diffusivity of substrate into the gel, and
density of the gel support.
Analysis:
We desire to know the maximum size of the particles to achieve a given effectiveness
factor. As we dont know the substrate concentration in the bulk fluid, we can assume that
we can adjust it at will. Such that we can assume it to be small enough, such that:
kM
=
SS
Then, we can approximate the effectiveness factor as:
3 1 1
h= 0.9
tanh( )
The Thiele modulus is given by:
mol
1/ 2
1 mmol g
250 1.1 3
min g 1000 mol
= R max = R
v cm
k M De mmol 2 cm
2
1L
58 5.1 10
L min 1000 cm 3
= 9.642 R
Then,
3 1 1
0.9 R 0.137 cm
9.642 R tanh(9.642 R ) 9.642 R
4. Known: ratio of initial reaction rate to initial bulk substrate concentration for two
different enzyme loads in the particles.
Analysis:
The reaction rate without diffusion limitation can be calculated using the Michaelis-
Menten expression with S equal to the bulk substrate concentration (S = S0):
vmax S 0 k [ E ]S
v (no diffusion) = = 2 0 0
kM + S0 kM + S0
Then,
k 2 [ E0 ]S 0
v (no diffusion, [ E0 ] = 10M) =
kM + S0
2k 2 [ E0 ]S 0
v (no diffusion, [ E0 ] = 20M) =
kM + S0
2k 2 [ E0 ]S 0
v (obs) 2
kM + S0 v (obs) 2
= =
k 2 [ E0 ]S 0 2v (obs)1
v (obs)1
kM + S0
Then,
v (obs) 2 1.00 S 0
= = = 0.769
2v (obs)1 2 0.65S 0
This means that doubling the enzyme concentration in the particles resulted in reduction
of the effectiveness factor to 76.9% the original effectiveness factor.
vmax k [E ]
=R =R 2 0
k M De k M De
Then,
2k 2 [ E 0 ]
R
2 k M De
= = 2 = 1.414
1 k 2 [ E0 ]
R
k M De