Chapter Three:

Enzymes

1
 Inhibition of Enzymes
Can be irreversible (metals) or
reversible (product, substrate, salt,
etc.)
Types:
1. Competitive
2. Noncompetitive
3. Uncompetitive
4. Substrate Inhibition
2
 Competitive Inhibition
Inhibitor is an analog of the substrate, and binds to the active
site of the enzyme.

3
Mechanism of Competitive Inhibition

[E ][S ]
E  S  ES  E  P K m 

[ES ]
I [E ][I ]
KI 
 [EI ]
EI
[E 0 ]  [E ]  [ES ]  [EI ]

v  k 2 [ES]
What assumption have we make in defining the 4
parameters on the right?
 Competitive Inhibition
Product formation rate is given by:

dP Vm [ S ] Vm [ S ]
v  
dt  [I ]   , app  [ S ]
Km
 1 
Km   [S ]
 KI 

What is the magnitude of K m  , app relative

to K m and what will be the effect on v?
How could you run a process to minimize the
effects of this type of inhibition?
5
 Competitive Inhibition
1/v I>0

I=0

Vm is unchanged
1/Vm

-1/Km -1/Km,app 1/[S]

6
 Noncompetitive Inhibition
Inhibitor binds to the enzyme and/or enzyme-substrate
complex, but not at the active site. However, the
enzyme affinity for substrate is reduced.

7
Mechanism of Noncompetitive
Inhibition

E  S  ES  P [E ][S ] [EI ][S ]

K m  
  [ES ] [ESI ]
I I
  [E ][I ] [ES ][I ]
KI  
 S  ESI
EI [EI ] [ESI ]


[E 0 ]  [E ]  [ES ]  [EI ]  [ESI ]

v  k 2 [ES]
8
 Noncompetitive Inhibition
Product formation rate is given by:
Vm Vm ,app
v 
 [ I ]   K m   K m 
1  K  1  [ S ]  1  [ S ] 
 I    

Question: What is the magnitude of Vm,app

relative to Vm, and what will be the effect on
v? How can you moderate the effects of this
type of inhibition. 9
 Noncompetitive Inhibition
1/v I>0

I=0

1/Vm,app

1/Vm

-1/Km 1/[S]
Km is unchanged 10
 Uncompetitive Inhibition
Inhibitor binds only to ES complex, and not
to E alone.
E  S  ES  P [E ][S ]
K m 
 [ES ]
I
[E ][I ]
 KI 
[EI ]
ESI

[E 0 ]  [E ]  [ES ]  [ESI ]

v  k 2 [ES]
11
 Uncompetitive Inhibition
Rate is given by:
Vm
[S ]
 [I ] 
1  
 KI  Vm, app [ S ]
v 
Km 
 [ S ] K m,app  [ S ]
 [I ] 
1  K 
 I

What is the magnitude of Vm,app relative to Vm? What

is the magnitude of  relative
Km , app to ?Km
12
 Uncompetitive Inhibition
1/v
I>0

I=0
1/Vm,app

1/Vm

-1/Km,app -1/Km 1/[S]

13
 Substrate Inhibition
Substrate binds to ES complex.

E  S  ES  P [E ][S ]
 K m 
[ES ]
S

[ ES ][ S ]
Ks 
ES 2 [ ES 2 ]
[ E0 ]  [ E ]  [ ES ]  [ ES 2 ]
v  k 2 [ES]
14
 Substrate Inhibition
No substrate inhibition
v

Substrate inhibition
Vm[ S ]
v
[ S ]2
K m  [ S ] 
K Si

S
The maximum substrate concentration resulting in maximum reaction rate is:

[ S ]m. rate  K m K Si 15
Summary of Enzyme Inhibition

16
Example 3.2
Example 3.2
Example 3.2
Example 3.3
Example 3.3