SCITAMA

ENZYMES

Enzyme Kinetics Defined

Study of rates of chemical reactions

Concentration of reactants and products in molarity
Can be determined either by:
 Rate of disappearance of a reactant
 Rate of appearance of a product

Solute

Made from substance

Typical Kinetic Experiment:

[E] = k >> vary [S] (S – Substrate) >> measure Vi

A (μM-1)
 ○ <<[S] μM (Micromolar) V
 S1 _ V1
 S2 _ V2
 S3 _ V3
 Sn _ Vn
The enzyme was diluted around 40 times (Add the volume) = Enzyme solution (1mL
saliva + 9mL H2O + 30mL 0.5% NaCl)

Simple Enzymes (Find Graph)

Reaction rate vs Substrate concentration

Hyperbolic
Binding of myoglobin to oxygen
Michaelis-Menten Kinetics – Described Hyperbolic Graph

Allosteric Enzymes (Find Graph)

Multi-subunit enzymes or oligomeric enzyme

Just like hemoglobin which is a tetrameric protein
Follow complex kinetics
Need to solve kinetic constants

Michaelis-Menten Plot (Find Graph)

Vi = initial velocity (moles/time)

[S] = substrate concentration (molar)
Vmax = maximum velocity
Km – substrate concentration when
 Vi is one-half Vmax

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 If the value of this is high, it has low affinity, it is loosely bound to the enzyme’s
active site and vice-versa
 (Michaelis-Menten constant)

Transforming the Michaelis-Menten Equation: Lineweaver-Burk Plot (Find Graph)

vv

vv

○
A

#30 – 7th Edition Book

Factors Affecting Enzyme Activity

Temperature (Find Graph – Percent maximum activity vs Temperature)

 Low temperature, the activity is low and vice-versa
 When optimum temperature is passed, the activity decreases
 Thermostable
 Comes from thermophilic bacteria (Hotsprings, Volcano, etc.)
 Optimal temperature goes beyond 90C
pH changes (Find 2 Graphs – Enzyme activity, Acidic, pH, Basic, Arginase, Pepsin, and
Salivary amylase || Cholinesterase, Papain, Pepsin, Chymotrypsin – relative activity vs
pH)
 Optimal pH is close to 7
 Above or below the optimal pH, the activity decreases
 Pepsin
 Stomach enzyme
 Optimal pH is 2
 Arginase
 Optimal pH is 10
Activators
Inhibitors
 Decrease the activity of enzymes
 Two groups:

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 Reversible inhibitors: (Find Graph)
 Binds noncovalently to the enzyme
 E + I <-> E – I
 Three types:
1. Competitive
 Inhibitor binds to the same site as the substrate
 “Substrate analogs” is its other name
 If it binds to the active site, there will be no reaction
 Vmax = same || Km =  || slow of L-B plot = 
2. Non-competitive
 Inhibitor binds to another site yet it affects the
active site because it causes a confirmational
change which affects the 3D structure of the active
site, therefore, the substrate can no longer bind as
efficiently and since the substrate can no longer
bind efficiently, it affects the activity of the enzyme
 Vmax =  || Km = same || slow of L-B plot = 
3. Uncompetitive (Find Graph
themedicalbiochemistrypage.org – Panel A, B, C, D – 1/v
vs 1/[S] – uninhibited enzyme, plus competitive inhibitor,
plus noncompetitive inhibitor, plus uncompetitive
inhibitor)
 Vmax =  || Km =  || slow of L-B plot = same
 Parallel
 Irreversible inhibitors:
 Bind covalently to the enzyme
 E + I -> E – I

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