A hyaluronidase from the Snake Venom of Agkistrodon Blomhoffii Ussurensis of Changbai Mountain: Isolation and Characterization

摘要

Snake venom is a complex mixture of protein and peptide toxins. Hyaluronidase a‘spreading factor’ has not been studied extensively in snake venom. In this paper, we describe the purification and characterization of a hyaluronidase from the snake venom of Agkistrodon blomhoffii ussurensis of Changbaimountain. Hyaluronidase has been purified by the successive chromatography on a DEAE-Sephadex A-25, on CM- Sephadex C-25 and on Sephadex G-75 columns on the basis of the purification method of thrombin-like enzyme (TLE). The molecular mass was found to be 35.0 kDa by SDS-polyacrylamide gel electrophoresis. Hyaluronidase was 5.66 percent of the snake venom of Agkistrodon blomhoffii ussurensis of Changbai mountain, the activity of mass (u/g) was 437218.5. The pH optimum for the enzyme was in the rang of 3.0-4.0 and 6.0-6.5,maximum activity was obtained at 37?, and the optimum concentration of Na+(mol/L) was in the rang of 0.3-3.0, the Km for hyaluronidase, using hyaluronic acid as substrate, was 4.633 × 10-3 mg/ml. The hyaluronidase had not the inhibition effect to growth of the Hela cell and HepG-2 cell in vitro through the experiment.