Related Experiment Videos
A procedure for urease and protein extraction from staphylococci.
C H Sissons1, E M Hancock, H E Perinpanayagam
1Dental Research Unit, Medical Research Council of New Zealand, Wellington.
The Journal of Applied Bacteriology
|October 1, 1989
Summary
A new method efficiently solubilizes staphylococcal urease and cell protein using phosphate buffer extraction. This simple technique recovers up to 100% of urease, aiding in staphylococcal research.
Area of Science:
- Microbiology
- Biochemistry
Background:
- Staphylococcal urease and protein extraction are crucial for research.
- Existing methods may be complex or inefficient.
Purpose of the Study:
- To develop a simple and effective method for solubilizing staphylococcal cell protein and urease.
- To compare the new extraction method with sonication.
Main Methods:
- Culturing staphylococci in Todd-Hewitt broth with yeast extract.
- Extraction of cells using phosphate buffer (pH 7.0) for 18-24 hours.
- Electrophoretic analysis of extracted proteins and urease.
Main Results:
- The phosphate buffer extraction method successfully solubilized staphylococcal urease and protein.
- Up to 100% of cellular urease was recovered, with generally 20% solubilized.
- Protein solubilization was less efficient than urease.
- Electrophoretic patterns from the new method matched those from sonication for Staphylococcus epidermidis.
Conclusions:
- The developed extraction procedure is simple and effective for isolating staphylococcal urease and protein.
- This method minimizes cell handling and offers comparable results to sonication.
- The technique is applicable to various staphylococcal species, including Staphylococcus aureus and coagulase-negative staphylococci.