Related Experiment Videos
[Purification and properties of staphylococcal hyaluronidase]
Abstract:
Microbial hyaluronidase (EC 4.2.2.1) was isolated from the culture fluid of Staphylococcus aureus 0-15 with purification by precipitation with 1 volume of ethyl alcohol, chromatography on DEAE cellulose and ultrafiltration through DA type membranes with the pore size of 0.65 micron ("Millipore") and PM-10 membranes ("Amicon"). The specific activity of the enzyme averaged to 2700 turbidimetric units or 32130 IU. 6585-fold purification of the enzyme was performed. The optimum action on hyaluronic acid was observed at pH 5.0-6.5. Hyaluronidase was inhibited by Fe3+, Fe2+ and Cu2+, activated by Ca2+ and stabilized by 0.15 M NaCl. It was detected that the enzyme had two molecular forms with the isoelectric points of 5.4 and 6.5 and the molecular weights of 55 000 and 24 0000 D respectively. The glycoprotein nature of the enzyme was shown. The immobilized form of hyaluronidase on activated polyglucin, a soluble biocompatible polymer was prepared. The form is characterized by higher thermostability.
Insights
Researchers purified microbial hyaluronidase from Staphylococcus aureus, achieving high specific activity and identifying optimal conditions for its action. This work provides insights into enzyme properties and immobilization strategies.
Area of Science:
- Microbiology
- Enzymology
Context:
- Hyaluronidase enzymes are crucial in various biological processes and biotechnological applications.
- Staphylococcus aureus is a known source of microbial enzymes with potential industrial relevance.
Purpose:
- To isolate and purify microbial hyaluronidase from Staphylococcus aureus 0-15.
- To characterize the biochemical properties of the purified enzyme, including optimal pH, stability, and molecular forms.
- To investigate the potential for enzyme immobilization.
Summary:
- Microbial hyaluronidase was purified from Staphylococcus aureus 0-15 using alcohol precipitation, DEAE cellulose chromatography, and ultrafiltration, achieving a 6585-fold purification.
- The enzyme exhibited optimal activity on hyaluronic acid at pH 5.0-6.5, was inhibited by Fe3+, Fe2+, and Cu2+, activated by Ca2+, and stabilized by 0.15 M NaCl.
- Two molecular forms with distinct isoelectric points and molecular weights were identified, and the enzyme's glycoprotein nature was confirmed. An immobilized form on activated polyglucin demonstrated enhanced thermostability.
Impact:
- The study provides a detailed characterization of Staphylococcus aureus hyaluronidase, contributing to the understanding of microbial enzyme diversity.
- The development of a thermostable immobilized hyaluronidase offers potential for improved applications in biotechnology and medicine.
- This research lays the groundwork for further exploration of microbial hyaluronidases in industrial and therapeutic settings.