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k1
A B (+ C)
Decay reactions, like radio-activity;
SN1 reactions
d[A] [A]
Rate: - = k1[A]
dt
d[A]
Rewriting: - = k1dt t
[A]
t t
Integration gives: 1 d[A] ktdt
0 [A] 0
[ A ]t
So: ln[A]t ln[A]0 = -kt or: ln = -kt
[ A]0
The time for half of the
reactant initially present to
decompose, its half-time or
half-life, t1/2,isaconstantand
hence independent of the
initial concentration of
reactant.
t1/2=ln2/k=0.693/k
2A P
Second-order reaction
The half-time for a second order reaction is expressed t 1/2 = 1/k [A]0 and therefore, in contrast to a
A+B P
When [S] [E] : the rate is zero order with respect to sucrose
The Michaelis-Menten Equation
2. under the physiologically common conditions that substrate is in great excess over
3. Enzyme ([S] [E]).
The expression of the initial velocity (v0) of the reaction, the velocity
at t=0, thereby becomes
The maximal velocity of a reaction, Vmax occurs at high substrate concentrations when
the enzyme is saturated, that is, when it is entirely in the ES form
The magnitude of KM varies widely with the identity of the enzyme and the nature of the substrate. It
is also a function of temperature and pH. The Michaelis constant can be expressed as
Since Ks is the dissociation constant of the Michaelis complex, as Ks decreases, the enzymes affinity
for substrate increases. KM in therefore also a measure of the affinity of the enzyme for its substrate,
provided k2/k1 is small compared to Ks, that is k2 k-1 so that the ES P reaction proceeds more
slowly than ES reverts to E + S
This quantity is also known as the turnover number of an enzyme because it is the number of
reaction processes (turnovers) that each active site catalyzes per unit time.
Turn Over Numbers of Enzymes
O R O
=
H3CCNCCOCH3
H H
R= kcat / Km
Glycine H 1.3 10-1
Norvaline CH2CH2CH3 3.6 102
(M-1 s-1)
Adapted from Mathews et al (2000) Biochemistry (3e) p.379
Al iniciar:
t = 0, S = So
A cualquier tiempo:
T=t S=S
X = (So-S)/So
- dS/dt = vi = So dX/dt
TemperatureDependenceofEnzymes
Asisthecasewithmostreactions,anincreasein
temperaturewillresultinanincreaseinkcatforan
enzymaticreaction.
Fromgeneralprinciples,itcanbedeterminedthat
therateofanyreactionwilltypicallydoublefor
every10Cincreaseintemperature.
Manyenzymesdisplaymaximumtemperatures
around40C,whichisrelativelyclosetobody
temperature.
Thereareenzymesthatareisolatedfrom
thermophilicorganismsthatdisplaymaximaaround
100C,andsomethatareisolatedfrom
psychrophilicorganismsthatdisplaymaximaaround
10C.
Enzyme Inhibition (Mechanism)
S S E I
S
E S I I
I
Compete for S I
Inhibitor active site Different site
E + S
ES E + P E + S ES E + P E + S
ES E + P
Equation and Description
+ + + +
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].
Succinate Dehydrogenase
Adapted from Kleinsmith & Kish (1995) Principles of Cell and Molecular Biology (2e) p.49
Sulfa Drug Is Competitive Inhibitor
Domagk (1939)
Folic Tetrahydro-
Precursor
acid folic acid
Many substances alter the activity of an enzyme by reversibly combining with it in a way
what influence the binding of substrate and/or its turnover number. Substances that
reduce an enzymes activity in this way are known as inhibitors
CompetitiveInhibition
A substance that competes directly with a normal substrate for an enzymes substrate-
binding site is known as a competitive inhibitor.
CompetitiveInhibition
CompetitiveInhibition
A plot of this equation is linear and has a slope of KM/Vmax, a 1/[S] intercept
of -1/ KM, and a 1/v0 intercept of 1/ Vmax
EnzymeInhibition
UncompetitiveInhibition
In uncompetitive inhibition, the inhibitor binds directly to the enzyme-substrate complex
but not to the free enzyme
In this case, the inhibitor binding step has the dissociation constant
The uncompetitive inhibitor, which need not resemble the substrate, presumably distorts
the active site, thereby rendering the enzyme catalytically inactive.
EnzymeInhibition
UncompetitiveInhibition
The double-reciprocal plot consists of a family of parallel lines with slope KM/Vmax, 1/v0
intercepts of /Vmax and 1/[S] intercept of -/KM
EnzymeInhibition
MixedInhibition(noncompetitiveinhibition)
A mixed inhibitor binds to enzyme sites that participate in both substrate binding and
catalysis. The two dissociation constants for inhibitor binding
Double-reciprocal plots
consist of lines that have
the slope KM/Vmax, with a
1/v0 intercept of /Vmax and
1/[S] intercept of -/ KM
Enzyme Inhibition (Plots)
Vmax Vmax
I I I
Almost all of these so called bisubstrate reactions are either transferase reactions in
which enzyme catalyzed the transfer of a specific functional group, X, from one of
the substrates to the other:
Sequential Reactions