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ENZYMES SPEED UP CHEMICAL REACTIONS
� Cofactors
� Coenzymes
� Holoenzyme
� Apoenzyme
How Enzymes Work?
Enzymes act
by lowering the free
energy of the transition
state
Enzymes speed up metabolic
reactions by lowering energy barriers
Induced by the
substrate
Lock and Key Model
An enzyme binds a substrate in a region called the active site
Product is released
Enzyme Kinetics
At pre-steady-state,
At equilibrium, [P] is low (close to zero
no net change of [S] & [P] time), hence, V0 for initial
or of [ES] & [E] reaction velocity
At pre-steady state, we can ignore the back reactions
Michaelis-Menten kinetics
(summary)
Enzyme kinetics (Michaelis-Menten Graph) :
At fixed concentration of enzyme, V0 is almost linearly proportional to
[S] when [S] is small, but is nearly independent of [S] when [S] is large
Proposed Model: E + S ES E + P
ES complex is a necessary intermedi
Objective: find an expression that relates rate of catalysis to the
concentrations of S & E, and the rates of individual steps
Start with: V0 = k2[ES], and derive, V0 = Vmax x[S]/([S] + Km)
This equation accounts for graph data.
At low [S] ([S] < Km), V0 = (Vmax/Km)[S]
At high [S] ([S] > Km), V0 = Vmax
When [S] = Km, V0 = Vmax/2.
Thus,
Range of Km values
Less E available,
V max is lower,
Km remains the same
for available E
Km unaltered, V max decreased
Uncompetitive Inhibitors
• ES + I = ESI
[ S ][ ES ] ' [ S ][ E ]
E+S
Km’
ES
k2
E+P K Si , Km
+ [ ES 2 ] [ ES ]
S
Vm [ S ]
v 2
KS1 [ S ]
K m' [ S ]
ES2
K S1
Substrate Inhibition
At low substrate concentrations [S]2/Ks1<<1 and
inhibition is not observed
1/V 1/V
I>0
I=0
1/Vm,app
1/Vm 1/Vm
S S I
E S I I
I
Compete for S I
Inhibitor active site Different site
Equation and Description
E + S→
← ES → E + P E + S←→ ES → E + P E + S←
→ ES → E + P
+ + + +
I I I I
↓↑ ↓↑ ↓ ↑ ↓↑
EI EI + S →EIS EIS
[I] binds to free [E] only, [I] binds to free [E] or [ES] [I] binds to [ES] complex
and competes with [S]; complex; Increasing [S] can only, increasing [S] favors
increasing [S] overcomes
not overcome [I] inhibition. the inhibition by [I].
Inhibition by [I].
Enzyme Inhibition (Plots)
Vmax’ Vmax’
I I I
- on substrate
k d Ad e Ea / RT
v Ae Ea / RT E0 e k d t
Factors Affecting Enzyme Kinetics
Temperature
- on the rate of enzyme catalyzed reaction
d [ P]
v k 2 [ ES ]
dt
k2=A*exp(-Ea/R*T)
T k2 v
- enzyme denaturation d[E ]
kd [E]
T Denaturation rate: dt
kd=Ad*exp(-Ea/R*T)
kd: enzyme denaturation rate constant;
Ea: deactivation energy
REFERENCES
Michael L. Shuler and Fikret Kargı, Bioprocess
Engineering: Basic Concepts (2 nd
Edition),Prentice Hall, New York, 2002.
www.biochem.umass.edu/courses/420/lectu
res/Ch08B.ppt -
class.fst.ohio-
state.edu/fst605/605p/Enzymes.pdf –
www.horton.ednet.ns.ca/staff/selig/powerpoint
s/bio12/biochem/enzymes.pdf
www.siu.edu/departments/biochem/som_pbl/S
SB/powerpoint/enzymes.ppt
www.associazioneasia.it/adon/files/2005_luisi_
05_why_are_enzymes.pdf
www.fatih.edu.tr/~abasiyanik/Chapter6_enzym
es.pdf -
http://www.authorstream.com/presentation/kkoz
ar-14001-enzymes-enzyme-ppt-education-powerpo
int/
http://highered.mcgraw-hill.com/sites/0072495
855/student_view0/chapter2/animation__how_enz
ymes_work.html
http://www.wiley.com/college/pratt/0471393878/s
tudent/animations/enzyme_kinetics/index.html