Catalase Test for Bacterial Identification
Translated by
ScribdTranslationsCatalase Test for Bacterial Identification
Translated by
ScribdTranslationsMicrobiology tests
CATALASE TEST
This test is the basis for the identification of gram-positive bacteria.
Reminders
Certain bacterial metabolic reactions result in aerobic production of hydrogen peroxide (water
(oxygenated). The spontaneous reoxidation by oxygen of the coenzyme FADH2 is its essential source:
FADH 2 + O2 FAD + H2O2
H2O2 can also be the product of the activity of superoxide dismutase, the superoxide ion resulting from the action of oxygen.
on various organic molecules, or that may be the product of the respiratory chain.
Hydrogen peroxide must be eliminated, as it is a cellular poison (it is used as an antiseptic). Its
decomposition in the microbial organism can be carried out either by peroxidases or by catalase. In the absence of a system
Destructive enzymatic action makes aerobic life generally impossible: the microorganisms then become strict anaerobes.
Note: Role of peroxidases
They catalyze the decomposition of hydrogen peroxide into water through the following redox reaction:
RH + H2 O ROH + 2 H+ + 2 e-
H2O2+ 2H+ + 2e- 2 H2O
RH + H2O2 ROH + H2O
Peroxidases are generally less active than catalase.
Principle
This enzyme is produced in abundance by bacteria with respiratory metabolism (AS and AAF) that can destroy
peroxides. Catalase is an enzyme that catalyzes:
2 H2O2 2 H2O + O2
Most aerobic microorganisms possess catalase, especially aerobic Gram-negative bacilli.
Absence is therefore an interesting identification criterion. For example, among aerobic Gram + shells, only the Streptococcaceae.
are catalase negative. Lactobacillus and Erysipelothrix are the only groups of non-sporulating Gram + aerobic bacilli observed.
catalase.
The role of peroxidases or catalases contained in peptones or in certain additives (blood...) in the media is
determinant to allow the aerobic development of catalase-negative bacteria such as Streptococcaceae.
Technique
• on a clean, dry slide, place a drop of hydrogen peroxide at 10 volumes,
• using a buttoned Pasteur pipette, add the bacterial inoculum
• observe immediately.
• at the contact of an isolated colony or on the slope of a culture in agar, deposit a drop of hydrogen peroxide
• observe immediately.
Lecture
- appearance of bubbles, gas release of oxygen: catalase +
- no bubbles: catalase –
Causes of errors: conducting the test on a broth containing catalase, from
from a blood agar that has catalase activity, bacterial suspension
insufficient, expired hydrogen peroxide.
Pierre-Yves Guillaume
Professor of biotechnology biological engineering