Enzyme activity

How fast an enzyme is working

Rate of Reaction

Rate of Reaction = Amount of substrate changed (or amount product formed)

in a given period of time.

 In most cases, actual molar enzyme

concentration is not known. We measure
amount of enzyme by its activity in terms
of enzyme unit of rate or a IU.
IU is also a way to monitor the enzyme
purification.
In our study the enzyme activity we
measure is more related to amount of
enzyme rather than its physiological
situation.

IU
One IU is defined as the amount of
enzyme that produces a mole
product per minute under defined
conditions.
The concentration of enzyme is
expressed as IU/mL
The specific activity (SA) is the
number of IU per mg protein.
As the enzyme is purified the SA
increases

Rate of Reaction

Enzyme activity

Variable you are looking at

Enzyme activity

Four Variables

Enzyme activity
Temperature
Four Variables

pH
Enzyme Concentration
Substrate Concentration

Rate of Reaction

Temperature

Rate of Reaction

Temperature

10

20

30

40

50

60

Temperature

5- 40oC
Increase in Activity

Rate of Reaction

40oC - denatures

0
<5oC - inactive

10

20

30

40

50

60

Temperature and Enzyme Activity

Enzymes are most active at an optimum temperature
(usually 37C in humans)
They show little activity at low temperatures
Activity is lost at high temperatures as denaturation
occurs

Effect of heat on enzyme activty

If you heat the protein above its optimal temperature

bonds break
eaning the protein loses it secondary and tertiary structure

Effect of heat on enzyme activty

Denaturing the protein

Effect of heat on enzyme activty

Denaturing the protein

ACTIVE SITE CHANGES SHAPE
SO SUBSTRATE NO LONGER FITS

Even if temperature lowered

enzyme cant regain its correct shape

Rate of Reaction

pH

Rate of Reaction

pH

1
2
3
4
5
6
7
8
9

pH
Rate of Reaction

Narrow pH optima

pH
Rate of Reaction

Narrow pH optima

WHY?

pH
Rate of Reaction

Narrow pH optima

Disrupt Ionic bonds - Structure

Effect charged residues at active
site

pH and Enzyme Activity

Enzymes are most active at optimum pH
Amino acids with acidic or basic side-chains
have the proper charges when the pH is
optimum
Activity is lost at low or high pH as tertiary
structure is disrupted

Optimum pH for Selected Enzymes

Most enzymes of the body have an optimum pH
of about 7.4
However, in certain organs, enzymes operate at
lower and higher optimum pH values

Rate of Reaction

Enzyme Concentration

Rate of Reaction

Enzyme Concentration

Enzyme Concentration

Enzyme Concentration and Reaction Rate

The rate of reaction increases as enzyme
concentration increases (at constant substrate
concentration)
At higher enzyme concentrations, more enzymes
are available to catalyze the reaction (more
reactions at once)
There is a linear relationship between reaction
rate and enzyme concentration (at constant
substrate concentration)

Rate of Reaction

Substrate Concentration

Rate of Reaction

Substrate Concentration

Substrate Concentration

Substrate Concentration
Rate of Reaction

Active sites full- maximum turnover

Substrate Concentration