The amount of enzyme which liberates one micromole of N-acetylglucosamine per minute at 37°C and pH 4.0.
Assay Method
The assay is based on the following reaction:
An anhydro sugar is first formed from N-acetylglucosamine, followed by the conversion of the sugar into its furan derivative, as a result of increasing the acidity of the solution. This furan derivative then reacts with p-dimethylamino-benzaldehyde to form a colored complex, which is measured spectrophotometrically at 585 nm. Detailed procedures for the assay of hyaluronidase are available in Methods of Enzymatic Analysis.
Applications
(Hyaluronate 4-glycanohydrolase; EC 3.2.1.35 ) Hyaluronidase catalyzes the depolymerization of mucopolysaccharides, hyaluronic acid, and the chondroitin sulfates A and C. The enzyme is widely distributed in animal tissues but is found in great concentrations in the bovine and ovine testes. It is also produced by a number of bacteria. Hyaluronidase from bovine testes has a molecular weight of 55,000. Purified hyaluronidase is used clinically for the intradermal administration of large volumes of fluid when intravenous injections are contraindicated. The enzyme is administered prior to or simultaneously with the fluid and it facilitates absorption of the fluid.
