Bio-technology General Corporation

Yaffa Beck

Jacob R Hartman

Benjamin Greenman

Abraham Nimrod

Dov Kanner

Moshe Landsberg

Gene encoding human manganese superoxide dismutase and recombinant polypeptide encoded thereby

Human manganese superoxide dismutase DNA, its expression and method of recovering human manganese superoxide dismutase

This invention provides plasmids for expression of human superoxide dismutase of analogs thereof. The invention further provides a method of producing enzymatically active human MnSOD or an analog thereof by introducing plasmids for expression of human MnSOD into procaryotic or eucaryotic cells and growing the resulting cells under suitable conditions, including conditions wherein the production medium is supplemented with Mn.sup.++. The invention additionally provides a method of recovering purified enzymatically active MnSOD from procaryotic and eucaryotic cells. Human MnSOD or analogs thereof may be used to reduce reperfusion injury, prolong the survival time of isolated organs or treat inflammations or bronchial pulmonary dysplasia.

Methods of use of human manganese superoxide dismutase

An expression plasmid has been constructed which includes DNA encoding human manganese superoxide dismutase. Such plasmids may be introduced into host cells and the resulting cells cultured or grown under suitable conditions so as to produce enzymatically active human manganese superoxide dismutase or analogs thereof which may then be recovered. A method of producing an enzymatically active analog of human manganese superoxide dismutase by supplementing the culture media with Mn.sup.++ has been developed.

Plasmids for expression and method of producing a human manganese

A double-stranded cDNA molecule which includes DNA encoding human manganese superoxide dismutase has been created. The sequence of one strand of a double-stranded DNA molecule which encodes human maganese superoxide dismutase has been discovered. Such molecules may be introduced in procaryotic, e.g., bacterial, or eukaryotic, e.g., yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions so as to produce human manganese superoxide dismutase or analogs thereof which may then be recovered. Human MnSOD or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs, or treat inflammations. The invention also concerns a method of producing enzymatically active human manganese superoxide dismutase or an analog thereof in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase by maintaining the bacterial cell under suitable conditions and in a suitable production medium. The production medium is supplemented with an amount of Mn.sup.++ so that the concentration of Mn.sup.++ in the medium is greater than about 2 ppm. This invention also concerns a method of recovering purified enzymatically active manganese superoxide dismutase from bacterial cells.

Polypeptide analogs of human manganese superoxide dismutase and

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 precipation, redissolving and reprecipating the hyaluronate. Composition of sodium hyaluronate which are characterized by an absence of pyrogenicity and skin irritation are obtained.

Growing community of inventors

Gadera, Israel

Yaffa Beck