CHAPTER 1: THE KINETICS OF ENZYME CATALYZED REACTIONS BY: EN. MOHD. FAHRURRAZI TOMPANG ERT 211/1 Biochemical Engineering
CHAPTER 1: THE KINETICS OF ENZYME
CATALYZED REACTIONS
BY:
EN. MOHD. FAHRURRAZI TOMPANG
ERT 211/1 Biochemical Engineering
Michaelis-Menten Kinetics
Enzyme E and substrate S combine to form a complex ES, which then dissociates into product P and free or uncombined enzyme E:
Michaelis-Menten Kinetics
Equilibrium Constant
Enzyme balance
Decomposition of the complex to product and free enzyme is irreversible.
Product formation Rate, v
Michaelis-Menten Kinetics
Solving the equations
Substitute e in following equation esee o
mKk
k
es
se
1
1
sK
sese
m 0
Michaelis-Menten Kinetics
From Product formation equation
sK
sek
m 0
2
sK
svv
m max
02max Where ekv
Quasi-steady-state Approximation
Briggs and Haldane first proposed Quasi-steady-state assumption
Applying mass balance for substrate and intermediate
esksekdt
dsv 11
eskksek
dt
esd211
Quasi-steady-state Approximation
In a batch reactor at closed system [E0] is considered very small compared S
Therefore, d(es)/dt ≈0
From equation
21
1
kk
sekes
eskksek
dt
esd211
Quasi-steady-state Approximation
Substituting e
Production formation kinetics
s
k
kk
sees
1
21
0
eskdt
dp
dt
dsv 2
skkk
sek
121
02
/
Quasi-steady-state Approximation
Substituting
There is difference between Michaelis-Menten and Quasi-steady-state constant.
02max ekv
1
21m and
k
kkK
sK
svv
m max
Evaluation of Parameters in Michaelis-MentenEquation
Lineweaver-Burk plots are convenient for determination of Km
Double reciprocal plot
plot v versus v/[S] gives a line of slope –Km and y-axis intercept of Vm
Eadie–Hofstee plot
Hanes–Woolf plot
Plot of [S]/v versus [S] gives line of slope I/Vm
and y-axis intercept of Km/Vm.
This plot is used to determine Vm more accurately.
Modulation and Regulation of Enzyme Activity
Chemical species other than the substrate can combine with enzymes to alter or modulate their catalytic activity.
Such substances are called modulators or effectors, may be normal constituents of the cell.
They enter from the cell's environment or act on isolated enzymes.
Modulation and Regulation of Enzyme Activity
• The combination of an enzyme with an modulator is chemical reaction
• Modulator can be fully reversible, partially reversible, or essentially irreversible.
• Examples of irreversible inhibitors include poisons such as cyanide ions, which deactivate xanthineoxidase,
• Nerve gases, which deactivate cholinesterases
(enzymes which are part of nerve transmission).
Modulation and Regulation of Enzyme Activity
Reversible modulation of enzyme activity is one control mechanism employed by the cell to achieve efficient use of nutrients.
The enzyme regulation involve interconnected networks of reactions with several control loops
Modulation and Regulation of Enzyme Activity
Example, five-step sequence for the biosynthesis of the amino acid L-isoleucine.
Regulation of this sequence is achieved by feedback inhibition:
Modulation and Regulation of Enzyme Activity
The final product, L-isoleucine, inhibits the activity of the first enzyme.
Thus, if the final product begins to build up, the biosynthesis process will be stopped
Modulation and Regulation of Enzyme Activity
• enzyme-substrate inhibitors systems classify by their influence on the Michaelis-Menten equation parameters vmax and Km
• Reversible inhibitors are termed competitive if their presence increases the value of Km but does not alter vmax The effect of such inhibitors can be countered or reversed by increasing the substrate concentration.
• On the other hand, by rendering the enzyme or the enzyme-substrate complex inactive, a noncompetitiveinhibitor decreases the vmax of the enzyme but does not alter the Km value.
Mechanisms of Reversible Enzyme Modulation
Many competitive inhibitors bear close relationships to the normal substrates. This are called substrate analogs.
It is thought that these inhibitors have the key to fit into the enzyme active site, or lock,
But the key is not quite right so the lock does not work; i.e., no chemical reaction results.
Mechanisms of Reversible Enzyme Modulation
For example, inhibition of succinic acid dehydrogenation by malonic acid:
The malonic acid can complex with succinic dehydrogenase, but it does not react
Mechanisms of Reversible Enzyme Modulation
How the sulfa-drug act against bacteria?
The action of one of the sulfa drugs, sulfanilamide, is due to its effect as a competitive inhibitor.
Mechanisms of Reversible Enzyme Modulation
Sulfanilamide is very similar in structure to p-aminobenzoic acid, an important vitamin for many bacteria.
By inhibiting the enzyme which causes p-aminobenzoic acid to react to give folic acid, the sulfa drug can block the biochemical machinery of the bacterium and kill it.
Some noncompetitive inhibition and is thought to be the dominant mechanism for noncompetitive inhibition and activation. These are called allostericcontrol
An enzyme which possesses sites for modulation as well as catalysis has consequently been named an allosteric enzyme.