Hyaluronidase COAS 9001-54-1



Hyaluronidase

CAS: 9001-54-1 Merck: 13,4777 Storage condition: −20°C Form: Lyophilized powder Color: White to light brown

InChI: InChI=1S/C18H14O7/c19-13(11-2-4-15-17(6-11)25-9-23-15)7-12(18(20)21)10-1-3-14-16(5-10)24-8-22-14/h1-6,12H,7-9H2,(H,20,21) InChIKey: PPEBBOHQEAQKSW-UHFFFAOYSA-N SMILES: C(O)(=O)C(C1=CC=C2OCOC2=C1)CC(C1=CC=C2OCOC2=C1)=O

Product Introduction

Hyaluronan sodium is a physiologically active substance widely present in animal and human tissues. It is a high-molecular-weight polysaccharide biomaterial formed by alternating units of N-acetylglucosamine and glucuronic acid. It features high viscoelasticity, plasticity and good biocompatibility. It exists in skin, joint synovial fluid, umbilical cord, aqueous humor and vitreous body.

Hyaluronidase is an enzyme extracted from mammalian testes. It appears as white or slightly yellow powder. It is soluble in water, and insoluble in ethanol, acetone and ether. It is odorless and has good stability. Its activity remains stable after heating at 42 °C for 60 min, and about 80% activity can be retained after heating at 100 °C for 5 min. It tends to lose activity in low-concentration aqueous solutions; gelatin or gum arabic can be added for protection. Fe²⁺ and Cu²⁺ show reversible inhibitory effects on the enzyme. Pb²⁺, Hg²⁺ and Ni²⁺ have no obvious influence on enzyme activity. In the presence of sodium chloride (0.15 mol/L) or protamine sulfate, the inhibitory effect of chondroitin sulfate B, heparan sulfate, keratan sulfate, heparin sodium and high-concentration hyaluronic acid on hyaluronidase can be reversed. Aqueous solution of hyaluronidase is unstable and should be prepared before use.

Production Methods

Method 1: Extraction from bovine and sheep testes

1. Shredding and extraction: Clean frozen or fresh sheep testes, remove the outer membrane, inner membrane and epididymis, then mince. Add the minced material into pre-prepared acetic acid solution, stir intermittently for full extraction at low temperature for 4 hours, then separate the slurry. Residues can be reused for secondary extraction.
2. Salting-out and filtration: Add ammonium sulfate into the slurry under stirring, allow full dissolution and standing, then filter with polyester filter bags. Repeat salting-out to obtain crude product.
3. Salting-out and dialysis: Dissolve the crude product in pre-cooled distilled water, add ammonium sulfate gradually, stand in cold storage, then siphon and filter. Combine the filtrate and add more ammonium sulfate. Collect the precipitate and dissolve it in pre-cooled distilled water for dialysis in phosphate-citrate buffer (pH 6.5) for 24 hours.
4. Depyrogenation and drying: Centrifuge the dialysate. Add trisodium phosphate and calcium acetate solution under stirring, adjust pH to 8.5 with sodium hydroxide, filter, then neutralize the filtrate to pH 7 with hydrochloric acid, and carry out freeze-drying to obtain refined enzyme powder.
5. Lyophilized formulation: Dissolve the refined powder in 5% hydrolyzed gelatin aqueous solution, adjust pH to 5–7, sterile filtration, aliquot and freeze-dry to get lyophilized finished products.

Method 2: Microbial fermentation

Prepare culture medium containing starch, cottonseed meal, wheat bran and dry yeast. Inoculate Streptomyces, ferment at 30 °C for 3 days. Defoam and concentrate the fermentation broth, precipitate with acetone to get crude product. Purify by column chromatography, freeze-dry to obtain refined product. Dissolve refined powder in 5% hydrolyzed gelatin solution, sterile filtration and freeze-drying for finished lyophilized products.

Safety Information

Safety statements: 22-24/25 WGK Germany: 3 TSCA: Listed HS Code: 35079090 Storage class: 11 (combustible solid) Hazard category: Respiratory sensitizer, Category 1