Extraction method for hyaluronidase

Abstract

The invention provides an extraction method for hyaluronidase. Acid liquid is adopted to extract and ammonium sulfate is adopted to salt out in order to obtain a hyaluronidase crude product; and a hyaluronidase competitive product is obtained by implementing dialysis, pyrogen treatment and freeze-drying for the crude product. The method chooses sheep testis as a raw material, and adopts the high-speed centrifugal extraction method to separate and purify the hyaluronidase, so that the titer of the hyaluronidase can achieve 340 iu / mg.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for extracting hyaluronidase from fresh animal testes. Background technique

[0002] Hyaluronidase, also known as hyaluronidase, is abundant in the testes of mammals. It can hydrolyze hyaluronic acid and significantly reduce its viscosity, which is beneficial for sperm to enter the egg during fertilization. It also exists in sperm, submandibular gland, bee venom, snake venom, skin, spleen, leech and lysosome of cells.

[0003] The stability of hyaluronidase is good, and the activity will not be lost when heated at 42°C for 60 minutes; 80% of the activity can be retained when heated at 100°C for 5 minutes. Cooling after heat inactivation can restore some vitality. The enzyme is still relatively stable at pH below 5 or above pH 8. It is easy to inactivate in low-concentration aqueous solution, but it can be protected by adding 0.2% or 0.5% gum arabic or 0.2% gelatin. Fe 2...