Catalase Test

By
Dr.Maleeha Mufti
 Catalase - An Extraordinary Enzyme
Catalase is a common antioxidant enzyme found in
almost all aerobic organisms (organisms exposed to
oxygen) at an optimum temperature of 37°C.

It catalyzes the decomposition of Hydrogen

peroxide (H2O2) to water and molecular oxygen. .

Catalyze : To modify, especially to increase, the rate of (a chemical reaction) by catalysis.

Catalase Structure
Hydrogen Peroxide (H2O2 )
A by-product of various metabolic processes, a powerful oxidizing
agent and potentially damaging to cells.

By preventing excessive H2O2 build up Catalase allows important

cellular processes which produce H2O2 as a byproduct to take place
safely.
Hydrogen Peroxide (H2O2 ) Structure
The equation of Catalase Reaction
Catalase and Bacteria
 Catalase and Bacteria

Catalase is found in many aerobic (oxygen-growing

bacteria).
This works as a protective mechanism for bacteria, as
some immune cells generate hydrogen peroxide to kill
infectious bacterial cells.
 Catalase Test
Test used to differentiate catalase producing bacteria (staphylococci)
from non-catalase producing bacteria (streptococci)

An organism is tested for catalase production by bringing it into

contact with a small amount of hydrogen peroxide.
Bubbles of oxygen are released if organism is a catalase producer.
Culture should not be more than 24 hours old
Procedure
Slant Method
Materials Required
• Reagent: 3% Hydrogen Peroxide solution
• Cultures: 24-48 hour tryptic soy broth cultures of bacteria
• Media: Tryptic soy agar
 Catalase test procedure

• 3% Hydrogen Peroxide
solution in a dark color
container
Catalase test : Overnight Culture of Organism
Equipments
 Glass Capillary Tube ( or sterile wooden stick )
 Test tubes
Test tube rack
Method
• Pour 2-3 ml of H202 solution into a test tube
• Using a glass capillary tube remove several colonies of the test
organism and immerse in the H202 solution
• Look for immediate bubbling
catalase test procedure -
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catalase test procedure -
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catalase test procedure -
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Results

• Active bubbling…….positive catalase test

• No bubbling………...negative catalase test
catalase test procedure -
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catalase test procedure -
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• Always have a positive and negative control alongwith test organisms
Positive control…… staphylococcus species
Negative control…..streptococcus species
Cautions
• Performing the test on slide is not recommended because of the risk
of contamination from active bubbling
• Care must be taken when testing an organism cultured on medium
containing blood because catalase is present in RBCs
Extreme care must be taken if a colony is taken from a blood agar plate.
Erythrocytes contain catalase, and a transfer of only a few blood cells
can give a false-positive reaction.
List of catalase positive micro-organisms

All Staphylococci
Pseudomonas aeroginosa
Aspergillus fumigatus
Candida albicans
Enterobacteriaceae (Klebsiella, Serratia)
Mycobacterium tuberculosis produces a heat-labile catalase
workable only at body temperatures.
Fresh Wound
Effects after applying Hydrogen Peroxide