Extracellular hyaluronidase from Streptomyces koganeiensis

A process for preparing hyaluronidase from Streptomyces koganeiensis ATCC 31394 comprising:

• a) fermenting Streptomyces koganeiensis ATCC 31394 and obtaining a supernatant;
• b) submitting the supernatant obtained from the fermentation of Streptomyces koganeiensis ATCC 31394 to weak cation-exchange chromatography and isolating a protein fraction with hyaluronidase activity;
• c) submitting the protein fraction with hyaluronidase activity from step b) to diafiltration and strong anion-exchange chromatography and isolating a protein fraction with hyaluronidase activity;
• d) submitting the protein fraction with hyaluronidase activity from step c) to strong cation-exchange chromatography and isolating a protein fraction with hyaluronidase activity;
• e) submitting the protein fraction with hyaluronidase activity from step d) to strong anion-exchange chromatography and isolating a protein fraction with hyaluronidase activity, thereby obtaining hyaluronidase consisting of:
• a) an N-terminal amino acid sequence set forth in SEQ ID NO:
• 1,
• b) a molecular weight of 21.6 kDa,
• c) an isoelectric point (pI) ranging from 4.4 to 4.8,
• d) a relative enzymatic activity equal to or higher than 40,000 I.U./mg, measured against hyaluronidase BRP from bovine testis EDQM, FIP, Hyaluronidase, H1115000 and
• e) a HPLC purity degree higher than 98%, wherein
• said hyaluronidase is stable in aqueous solution and not sensitive to the action of proteolytic enzymes.