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Isolation and characterization of hyaluronidase from Streptococcus uberis
P Schaufuss1, R Sting, W Schaeg
1Institut für Bakteriologie und Immunologie, Fachbereich Veterinärmedizin, Justus-Liebig-Universität Giessen.
Zentralblatt Fur Bakteriologie : International Journal of Medical Microbiology
|May 1, 1989
Summary
Streptococcus uberis produces hyaluronidase, an enzyme crucial for bacterial virulence. This study details the enzyme's purification and characterization, revealing its optimal activity conditions and cofactor requirements for potential therapeutic targeting.
Area of Science:
- Microbiology
- Enzymology
- Biochemistry
Background:
- Streptococcus uberis is a significant mastitis pathogen in dairy animals.
- Hyaluronidase is a key virulence factor produced by various bacteria, including S. uberis.
- Understanding hyaluronidase properties is essential for developing targeted interventions.
Purpose of the Study:
- To isolate and purify hyaluronidase from Streptococcus uberis.
- To characterize the biochemical properties of the purified enzyme.
- To investigate factors influencing hyaluronidase activity.
Main Methods:
- Ammonium sulfate precipitation for initial isolation.
- Chromatography (DEAE-cellulose, ultragel ACA44) and isoelectric focusing for purification.
- Enzyme activity assays to determine optimal conditions and kinetic parameters.
Main Results:
- Purified hyaluronidase exhibited an isoelectric point of pH 4.9 and a molecular weight of approximately 54,000 D.
- Optimal enzyme activity was observed at pH 6.0 and 45°C.
- Michaelis constant (Km) was determined to be 7.0 x 10^-2 mg/ml.
- Enzyme activity was modulated by various divalent and monovalent cations.
Conclusions:
- Streptococcus uberis consistently produces hyaluronidase.
- The purified enzyme possesses distinct biochemical characteristics.
- Cationic modulation of hyaluronidase activity suggests potential targets for controlling bacterial spread.