CHONDROITINASE PRODUCTION IN RECOMBINANT PROTEUS VULGARIS STRAINS

摘要

Plasmids pLP<2>-1531, which encodes chondroitinases I and II, and pLP<2>-1521, which encodes chondroitinase I, are provided. These plasmids are used to transform wild-type E. coli and P. vulgaris cells. The transformed cells produce DNA encoding the production of chondroitinases I and II or chondroitinase I in the absence of natural exogenous chondroitinases I and II inducers. Additionally, the transformed P. vulgaris cells in the absence of exogenous chondroitinases I and II inducers, produce the respective enzymes in amounts in excess of those produced by wild-type P. vulgaris in the presence of exogenous chondroitinases I and II inducer(s). Starting plasmid pLP<2>-751 and intermediate plasmids pLP<2>-770, pLP<2>-1512, pLP<2>-1263, pLP<2>-1508, pLP<2>-1510, pLP<2>-1514, pLP<2>-1518, and pLP<2>-1525 are also provided. These intermediate plasmids are typically used in the preparation of pLP<2>-1531 and pLP<2>-1521. A BglII/EcoRI fragment of about 3970 bp derived from pLP<2>-1512, a EcoRI/SmaI fragment of about 2550 bp derived from pLP<2>-1514, and a BglII/SmaI(AvaI) fragment of about 6520 bp derived from pLP<2>-1518 are further disclosed. Also contemplated is the use of transformed P. vulgaris cells to produce chondroitinase I and/or II. The P. vulgaris cells above are cultured in a bacterial culture medium and in the absence of an exogenous chondroitinase I and II inducer. The cells are then harvested from the culture, and the chondroitinase I and/or II enzymes are recovered.