Enzymes

Define enzyme:
o It is a biological catalyst made of protein that speeds up rate of metabolic
reactions without being used up.
Their job:

To lower activation energy (energy needed to start chemical reaction)

 Breakdown large molecules into smaller ones

Nutrition

 Substrate:
o Substance on which enzyme acts on
 Product:
o Final product after reaction
 Active site:
o Has specific shape complementary to shape of enzyme’s specific
substrate.
==============================================================================
Properties of all enzymes:

o Biological catalyst
o Made of protein
o Specific (catalyses one reaction)
o Inactive by high temperature (work best at particular temperatures)
o Work best at particular ph
o Can be used again and again
Affected by:

 Temperature
 p.H
 Volume of substrate
 Volume of enzyme
 Concentration of substrate
 Concentration of enzyme
==============================================================================
Mechanism of action (5 marks)
Each enzyme has a specific shape of active site that acts on specific substrate. The shape of
substrate is complementary to shape of enzyme’s active site. So, the substrate binds into
active site of enzyme according to the key-lock mechanism (enzyme acts as lock and substrate
acts as key), to form the enzyme substrate complex, resulting in enzyme lowering activation
energy of reaction to catalyse reactions and help substrate to be broken down to form
products without being changed.
Effect of temperature on enzyme activity

DESCRIBE THE GRAPH

 As temp increases rate of reaction increases to a
point where it peaks then drops

EXPLAIN WHY?

 Lower temperatures:
o Enzyme keeps working but at a slower rate due to lower kinetic energy so less
effective collisions between enzymes and substrate
 Increase in rate:
o Why? This is because substrate and enzyme molecules gain more kinetic
energy so move faster so frequency of effective collisions increase and so does
the rate of reaction increase.
 Peak: Here enzyme is most active so rate of reaction is greatest and give highest yield
 Decrease in rate: Above 40 degrees enzyme molecules begin to denature (due to the
break down of cross links that keep the enzyme shape) and so it alters its active site
shape and prevents substrate from fitting.
o By 60 degrees the enzyme molecules are completely destroyed and reaction
stops causing it to be denatured.
Explain the effect of pH on the rate of reaction:
Each enzyme has an optimum pH, at which it is most active, the enzyme activity decreases
below or above the optimum pH, molecules are denatured at extremes of pH by destroying
cross links that hold enzyme’s shape together to allow substrate to fit into it.
USE UNIVERSAL INDICATOR TO RECORD PH IN
THE TEST TUBES
Investigation on effect of pH on rate of reaction of Catalase enzyme
Enzyme: Catalase
Substrate: Hydrogen peroxide (H2O2)

Rection: 2H2O2  2H2O + O2

Independent variable : pH Using Buffer
Dependant: Rate of catalyse
Control experiment: Same procedure but using denatured boiled enzyme
Control variables:

o Same concentration of hydrogen peroxide and enzyme

o Same volume of hydrogen peroxide and enzyme
o Same temperature Using thermostatically stable water bath
Methodology:

In a test tube pour 10cm3 of hydrogen peroxide and pour 5cm3 of catalyse enzyme
Place test tube in thermostatically stable water bath
Connect delivery tube to test tube containing solution and place it in an inverted test tube
submerged fully in water and is also connected to a gas syringe that collects oxygen gas
displaced.
To measure rate:
Volume of gas given off / Set time

 Repeat same experiment 3 times at each pH

 The one that gives the highest rate is the optimum pH

Other methodology:
o Count number of bubbles given off
 One with more bubbles is higher rate
This is inaccurate because: Some bubbles have different sizes and volumes
Also, we might miscount bubbles

For temperature just do same procedure but with different temperatures