Separation and Properties of Hyaluronidase From Ram Sperm Acrosomes

Summary. Ram sperm acrosomes were disrupted by Hyamine and Triton treatment and extracts were initially fractionated with a Sephadex G-100 column at pH 3·5. Acrosin, a proteolytic enzyme, was completely separated from hyaluronidase by this step. Highly active hyaluronidase, specific activity of 1320 units/mg protein (38,280 National Formulary units/mg protein), was obtained by DEAE chromatography but two minor contaminants were present. The partially purified enzyme had an optimum at pH 4·3. The enzyme showed no activity at pH 3·0 and only 10% of its activity at pH 8·0. Heparin and chondroitin sulphate B inhibited 50% of its activity. The molecular weight was estimated to be 62,000 by sodium dodecyl sulphate gel electrophoresis.