Abstract A novel mutant microorganism Streptococcus zooepidemicus HA-116 ATCC 39920, has been produced. The microorganism produces large amounts of high molecular weight hyaluronic acid. The invention provides a method of obtaining such microorganisms. The invention also concerns a method of obtaining sodium hyaluronate which comprises growing with vigorous agitation a microorganism of the genus Streptococcus under appropriate conditions in a suitable nutrient medium containing a sugar component as a carbon source. The sugar component is present in a substantially constant concentration between 0.2 and 10 grams per liter. The medium has a substantially constant pH between about 6.0 and 7.5 and includes a substantially constant magnesium ion concentration above 0.05 grams per liter. The sodium hyaluronate excreted into the medium by the organism is purified using methods involving precipitation, redissolving and reprecipitating the hyaluronate. Composition of sodium hyaluronate which are characterized by an absence of pyrogenicity and skin irritation are obtained.
A composition characterized by an absence of pyrogenicity and inflammatory activity and by the absence of proteins of animal origin which comprises between about 88% to 92% by weight sodium hyaluronate and from about 8% to 12% by weight water, the sodium hyaluronate being characterized by a minimum viscosity of about 3.5 m 3 /Kg, an average molecular weight from about 3.5×10 6 daltons to about 4.0×10 6 daltons, a glucoronic acid to N-acetyl glucosamine ratio of 1:1, from about 4% to about 6% sodium ion, a specific optical rotation measured at 25° C. and at a wavelength of 436 nm from about 155° to 165°, less than 0.01% by weight protein, less than 0.001% by weight sulfate, less than 0.02% by weight neutral sugar, less than 0.05 ng/ml of endotoxin, less than 0.2 mg/g of iron and less than 0.2 mg/g of copper.
Claim 2
Dependent Claim
144 words
A composition characterized by an absence of pyrogenicity and inflammatory activity and by the absence of proteins of animal origin which comprises between about 88% to 92% by weight sodium hyaluronate and from about 8% to 12% by weight water, the sodium hyaluronate being characterized by a minimum viscosity of about 3.5 m 3 /Kg, an average molecular weight from about 3.5×10 6 daltons to about 4.0×10 6 daltons, a glucoronic acid to N-acetyl glucosamine ratio of 1:1, from about 4% to about 6% sodium ion, a specific optical rotation measured at 25° C. and at a wavelength of 436 nm from about 155° to 165°, less than 0.01% by weight protein, less than 0.001% by weight sulfate, less than 0.02% by weight neutral sugar, less than 0.05 ng/ml of endotoxin, less than 0.2 mg/g of iron and less than 0.2 mg/g of copper.