Isolation and Partial Characterization of Hyaluronidase and Exo-polysaccharidases in Rat Carrageenin Granuloma

Abstract

Hyaluronidase, N-acetylglucosaminidase and β-glucuronidase of rat carrageenin granuloma were isolated and partially characterized, and changes in these enzyme activities during development of granuloma were also examined in order to clarify the role in metabolism of mucopolysaccharides of the granulomatous tissue. The exo-polysaccharidases, N-acetylglucosaminidase and β-glucuronidase, were present in both the granuloma and the pouch fluid, but hyaluronidase was dected only in the pouch fluid. After the gel filtration of pouch fluid on Sephadex G-200, hyaluronidase was separated completely from the two exo-polysaccharidases, and its enzymatic properties were examined. Optimum pH for the activities of hyaluronidase and N-acetylglucosaminidase was different from that of β-glucuronidase. The hyaluronidase fraction obtained was capable of degrading macromolecular hyaluronate and this fraction also depolymerized chondroitin 4-sulfate and 6-sulfate, but not dermatan sulfate. Appearance of these enzymes in the pouch fluid was periodically independent of one another ; maximum hyaluronidase activity was shown at 8 days after carrageenin injection and at initial experimental stage, higher N-acetylglucosaminidase was detected. On the other hand, β-glucuronidase was almost constant during the experimental period. The degradation of hyaluronic acid and the granuloma slices by hyaluronidase was not increased by the addition of N-acetylglucosaminidase and β-glucuronidase.