IIPE Class 4 24.03
IIPE Class 4 24.03
IIPE Class 4 24.03
Enzymes (History)
They often come from vitamins (organic nutrients required in small amounts in the
diet)
How enzymes work
• Enzymes provide specific environments in which
chemical reactions that don’t normally proceed under
neutral pH, mild temperature, and aqueous
environment conditions can occur
• This region is a pocket on the enzyme known as the
active site
• The molecule that is bound to the active site and
acted upon by the enzyme is called the substrate
• The two together form what is known as the enzyme-
substrate complex
• The function of an enzyme catalyst is to increase the
rate of a chemical reaction, not affect is equilibrium
• Therefore, enzymes don’t make more product, they
just make product faster
Active Site
• The area of an enzyme that binds to the substrate
• Structure has a unique geometric shape that is designed to
fit the molecular shape of the substrate
• Each enzyme is substrate specific
• Thus the active site that is complementary to the geometric
shape of a substrate molecule
Complementarity
• Geometric
• Electronic (electrostatic)
• Stereospecificity (enzymes and
substrates are chiral)
(1) Binding to the substrate can be done such that the formation
of the transition state is favored
(2) Orientation and positioning of substrate(s)
(3) Bonds in the substrate can be ‘activated’ by functional groups
in the catalytic site
Enzyme Kinetics
¾¾ ®
k1
E + S ¬¾¾ ES ¾¾® E + P
k2
k -1
d P
k 2 ES
[ES] is the difference between the ES
v formation and its disappearance.
dt
d ES
1
k 1 E S
Ks
k1 ES
Ks is the dissociation constant for the ES complex.
Assumption of steady state
Transient phase where in the course of a reaction the
concentration of ES does not change
d ES
0
dt
ET E ES 3
Combining 1 + 2 + 3
The KM
k 1 k 2 k2
can be expressed as: KM Ks
k1 k1 k1
Lineweaver-Burk Plot
1 = Km 1 1
1/V0
+
V0 Vmax [S] Vmax
Slope = Km/Vmax
-1/Km
1/Vmax
0 1/[S]
Lineweaver-Burk plot: slope = KM/Vmax,
1/vo intercept is equal to 1/Vmax
the extrapolated x intercept is equal to -1/KM
For small errors in at low [S] leads to large errors in 1/vo
• Reversible inhibition
reversibly bind and dissociate from enzyme,
activity of enzyme recovered on removal of
inhibitor - usually non-covalent in nature
– Competitive
– Uncompetitive
– Noncompetitive (Mixed)
• Irreversible inhibition
inactivators that irreversibly associate with
enzyme
activity of enzyme not recovered on removal -
usually covalent in nature
Competitive Inhibition
• Vo = Vmax[S]/(KM + ’[S])