Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1992-12-22
1995-05-23
Wax, Robert A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435691, 435219, 43525233, 4353201, 435273, 435 14, 435 66, 435 72, 435 73, 536 732, 536 237, 424 9467, C12N 952, C12N 1557, C12N 1570, C12N 1574
Patent
active
054181577
ABSTRACT:
Genetically engineered E. coli carry vectors containing inserts that code for Clostridium histolyticum collagenase. These inserts code for: (a) a form of collagenase having a molecular weight of about 68,000 daltons in the essential absence of larger forms of collagenase; (b) the 68 kd form of collagenase and a fusion polypeptide consisting of the collagenase protein fused to at least a portion of the .beta.-galactosidase protein of E. coli or (3) the 68 kd form of collagenase and polypeptides of molecular weight of from above about 68,000 daltons to about 100,000 daltons and having the enzymatic activity of C. histolyticum collagenase as indicated by digestion of .sup.3 H-acetylated collagen and by specific inhibition by 1,10-phenanthroline plus EDTA. The collagenase genes in the transformed E. coli are expressed efficiently in the transformed cells to yield enzymatically active and immunologically cross-reactive collagenase. In particular, the 68 kd form of collagenase is resistant to autocatalytic degradation and is stable to long-term storage. Genetically engineered collagenase, especially the 68 kd form that is resistant to autocatalytic degradation, can be used for isolation of pancreatic islets, for the isolation of dispersed tumor cells, or for treatment of "slipped disc."
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Lei Shau-Ping
Lin Hun-Chi
Brotman Harris F.
Moore William W.
Wax Robert A.
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