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Catalase is an enzyme that breaks down hydrogen peroxide into water and oxygen, primarily found in aerobic and facultative bacteria. The catalase test differentiates between bacteria, particularly identifying Micrococcaceae as positive and Streptococcaceae as negative. The test involves adding hydrogen peroxide to a bacterial culture and observing for bubble formation, with strict quality control measures in place using known positive and negative controls.

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Catalase is an enzyme that breaks down hydrogen peroxide into water and oxygen, primarily found in aerobic and facultative bacteria. The catalase test differentiates between bacteria, particularly identifying Micrococcaceae as positive and Streptococcaceae as negative. The test involves adding hydrogen peroxide to a bacterial culture and observing for bubble formation, with strict quality control measures in place using known positive and negative controls.

Uploaded by

Akbar Saleem
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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What is Catalase?

Catalase is an enzyme that decomposes hydrogen peroxide (H2O2) into water


and oxygen.
Chemically, catalase is a hemoprotein, similar in structure to hemoglobin,
except that the four
iron atoms in the molecule are in the oxidized (Fe3+), rather than the reduced
(Fe2+), state.
Excluding the streptococci, enterococci, and a few other genera, most aerobic
and facultative
bacteria possess catalase activity.

Principle in Catalase Test?


Hydrogen peroxide forms as one of the oxidative end products of aerobic
carbohydrate
metabolism. If allowed to accumulate, it is lethal to bacterial cells. Catalase
converts hydrogen
peroxide into oxygen and water as shown by the following reaction:
2H2O2 → 2H2O+ O2 (gas bubbles)

Use of Catalase Test?


The catalase test is most commonly used to differentiate members of the
Micrococcaceae,
which are positive, from members of the Streptococcaceae.

Reagent used in Catalase Test?

Hydrogen peroxide 3% stored in a brown bottle under refrigeration


An 18 to 24-hours culture of the organism to be tested

Quality Control in Catalase Test?


The hydrogen peroxide reagent must be tested with positive and negative
control organisms
each day or immediately before unknown bacteria are tested.
A. Positive control: Staphylococcus aureus
B. Negative control: Streptococcus species
Procedure of Catalase Test?
1. With an inoculating needle or a wooden applicator stick, transfer growth
from the center of
a colony to the surface of a glass slide.
2. Add one drop of 3% hydrogen peroxide and observe for bubble
formation.

Interpretation of Catalase Test?


The rapid and sustained appearance of bubbles or effervescence constitutes a
positive test.
Because some bacteria possess enzymes other than catalase that can
decompose hydrogen
peroxide, a few tiny bubbles forming after 20–30 seconds is not considered a
positive test.
In addition, catalase is present in red blood cells; so care must be taken to
avoid carryover
of red blood cells with the colony material.
CHART 1-2

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