Decarboxylases are a group of substrate-specific enzymes that are capable of reacting with the carboxyl (COOH) portion of amino acids, forming alkaline-reacting amines and byproduct carbon dioxide. Increased pH of the medium is detected by a color change of the pH indicators bromocresol purple and cresol red.
Bromcresol purple turns purple at an alkaline pH and turns yellow at an acidic pH.
Each decarboxylase enzyme is specific for an amino acid. Lysine, ornithine, and arginine are the three amino acids routinely tested in the identification of Enterobacteriaceae.
The specific amine products are:
- Lysine- Cadaverine
- Ornithine-Putrescine
- Arginine- Citrulline
These byproducts are sufficient to raise the pH of the media so that the broth turns purple. The medium used is arginine dihydrolase broth which is a nutrient broth supplemented with 0.5% arginine.
Arginine is hydrolyzed to ornithine (arginine is first converted to citrulline via dihydrolase reaction, in which NH2 group is removed from arginine. Citrulline is next converted to ornithine). Ornithine then undergoes decarboxylation to form putrescine. The production of amine, putrescine, elevates the pH, and the pH indicator Bromo cresol purple gives purple color in the medium (alkaline condition).
If the inoculated medium is yellow, or if there is no color change, the organism is decarboxylase-negative for that amino acid. If the medium turns purple, the organism is decarboxylase-positive for that amino acid.
Requirements
- Moeller decarboxylase base-4 tubes with lysine, ornithine and agrinine hydrochloride 1% and control
Method
- Inoculate the test medium, overlaid with sterile paraffin oil (Inoculated tubes must be protected from air to avoid false alkalinization at the surface of the medium)
- Incubate and read daily for four days
Result
- Purple color-Positive decarboxylation
- Yellow color-Negative i.e. no decarboxylation
Control
- Lysine: Klebsiella pneumoniae
- Ornithine: Enterobacter cloacae
- Arginine: Enterobacter cloacae
Negative
- Lysine: Enterobacter cloacae
- Ornithine: Klebsiella pneumoniae
- Arginine: Klebsiella pneumonia
Uses of decarboxylation Tests:
Arginine decarboxylase Test: It aids in differentiating enteric bacteria with closely related physiological characteristics.
Lysine Decarboxylase Test (LDC): To assist in the identification of Salmonellae (+ve) and Shigellae (-ve). Bacteria that are Lysine Decarboxylase (LDC) positive are:
- Escherichia coli
- Salmonella typhi and most other salmonellae species (except Salmonella paratyphi A).
- Klebsiella pneumoniae
- Serratia marcescens
- Vibrio cholerae
- Vibrio Parahemolyticus
References
- Clinical Microbiology Procedures Handbook, Fourth Edition. (2016). In Clinical Microbiology Procedures Handbook, Fourth Edition. American Society of Microbiology. https://doi.org/10.1128/9781555818814
- Color Atlas and Textbook of Diagnostic Microbiology, Koneman, 5th edition
- Ukimori, H., Kasahara, K., Shibata, K., & Moriyasu, T. (1971). Rinsho byori. The Japanese journal of clinical pathology, 19, 445.
in decarboxylase test bromocresol purple changes color (purple and yellow) then what is the use of cresol red
Thank u for sharing what u have!
thank u for ur informations….why we are use two indicators fpr this test
Hi, I’d appreciate if you add pictures for this experiment. Thanks : )
Thank you for all the very helpful information, and especially for the positive and negative examples! I always love using this site to get refreshers on various clinical microbiology topics and biochemical testing! 🙂
Thank you so much for your inspiring words. Let me know, how can i improve the overall user experience and usefulness of this site?
Pl include Falkows medium in your discussion,also add a point on acidic conditions produced through the fermentation of glucose that further triggers the production of decarboxylases.
Thus the change in colour of BCP from purple to yellow due to acids produced and further to purple because of decarboxylation and production of amines that are strongly alkaline.