CLL780 Tutorial 1

1.
a. Derive the Michaelis-Menten equation (MME) for enzyme kinetics. State
all the assumptions and their justifications.
b. What is the difference between quasi-equilibrium and steady state
approximation assumption? When these assumptions are applicable?
c. Explain the physical significance of various constants which appear in
MME.
2.
a. What are various linear representations of MME?
b. Derive them their respective linear expressions from M-M equation.
Mention their drawbacks.
c. What are the benefits of linearization? Do they still hold true when
compared to fitting data into original MME? (Hint: non-linear regression)
3.

a. What are various kinds of reversible enzyme inhibition? When they do occur?
(e.g. competitive inhibition occurs when the inhibitor resembles the
substrate)
b. Give their reaction mechanisms.
d.
c. Derive modified MMEs for competitive, uncompetitive, non-competitive and
mixed inhibition. Assume quasi-equilibrium.
d. How do Km app and Vmax app vary in each case? Explain their physical
significance. [Hint: problems 1c and 3a]
e. How do their Lineweaver-Burk plots vary? Explain using graphs.

4. The following data have been obtained for an enzyme.

Partial pressure of O2 O2 uptake rate (L O2 O2 uptake rate (L O2

(mmHg) /hr.mgcells) [without /hr.mgcells) [with
sulfanilamide] sulfanilamide]
0 0 0
0.5 23.5 17.4
1.0 33.0 25.6
1.5 37.5 30.8
2.5 42.0 36.4
3.5 43.0 39.6
5.0 43.0 40.0

a. Calculate Vmax, Km and turnover number for an uninhibited reaction.

b. If sulphanilamide added to the solution of is 20 mg/mL, establish which
kind of reversible inhibition is this. Find modified Vmax and Km.
c. Show corresponding Lineweaver Burk plot (only qualitative)