title: Staphylococcal hyaluronate lyase: purification and characterization studies.
abstract: Staphylococcal hyaluronate lyase (hyaluronidase) derived from a pathogenic strain of staphylococcus was purified by means of salt fractionation with ammonium sulfate and gel filtration through Sephadex G-100. Most of the enzyme activity from concentrated culture supernatant fluids of staphylococci was obtained in a fraction precipitated by 90 to 100% saturation with ammonium sulfate. A small amount of enzyme was also precipitated by 80 to 90% saturation with the salt. The hyaluronidase-rich fractions did not contain other staphylococcal enzymes, such as coagulase, protease, lipase, and staphylokinase. These enzymes were present in the original concentrates. Molecular sieving chromatography of the partially purified enzyme by filtration through Sephadex G-100 resulted in a further increase in specific enzyme activity. However, more than one active peak was obtained after gel filtration, thus suggesting that there may be more than one molecular form of the enzyme. Immunodiffusion in agar gel of the chromatographically purified enzyme fraction, with immune serum from rabbits injected with concentrated staphylococcal culture supernatant fluids, indicated that there was one major antigen. A similar antigen, giving reactions of identity with the purified material, was present in the original culture supernatant fluid.
mesh_terms: D002623:Chemistry Techniques, Analytical; D002850:Chromatography, Gel; D003030:Coagulase / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D005341:Fibrinolysin / Q000378:metabolism; D006460:Hemolysin Proteins / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D008049:Lipase / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D000644:Quaternary Ammonium Compounds; D013210:Staphylococcus / Q000201:enzymology*; D013696:Temperature
chemical_list: D003030:Coagulase; D006460:Hemolysin Proteins; D000644:Quaternary Ammonium Compounds; D003851:Deoxyribonucleases; D008049:Lipase; D010744:Phosphoric Monoester Hydrolases; D006821:Hyaluronoglucosaminidase; D010447:Peptide Hydrolases; D005341:Fibrinolysin