Simplified Two-Step Purification of Hyaluronidase from Buthus martensii Manchurian Scorpion Venom

Whole venom of Buthus martensii Karsch Manchurian scorpion was collected by electrostimulation and was fractionated by cation-exchange chromatography over Toyopearl CM650-M followed by ultrafiltration through an Amicon Ultra-15 membrane with a cut-off threshold of 10 kDa to isolate hyaluronidase. The specific activity of the purified hyaluronidase was 16,000 TRU/mg. The degree of purification calculated from the increase of specific activity was 71×; the yield, 75.6%. The electrophoretic and chromatographic purities were 88.4% and 82.5%, respectively.