http://rdf.ncbi.nlm.nih.gov/pubchem/patent/EP-1105515-A4
Outgoing Links
| Predicate | Object |
|---|---|
| assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_f8c0982b0f02b1ed6d20a315370a3a3a |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N9-1081 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12P19-18 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N9-10 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12P19-18 |
| filingDate | 1999-08-10-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_7d61e129e5c68f18b7e3e0acb3e383de http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_c802caa7880638896a4f756a0d7efab2 http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_8c5f1a8c1516d2e89ae04b83f73c15ff |
| publicationDate | 2002-08-28-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | EP-1105515-A4 |
| titleOfInvention | ALPHA 2.8 / 2.9 POLYSIALYLTRANSFERASE |
| abstract | The capsular polysaccharide of Escherichia coli K92 contains alternating (-8-NeuAc alpha 2-) and (-9-NeuAc alpha 2-) linkages. The enzyme catalyzing this polymerizing reaction has been cloned from the genomic DNA of E. coli K92. The 1.2 kb PCR<1> fragment was subcloned in pRSET vector and the protein was expressed in the BL21(DE3) strain of E. coli with a hexameric histidine at its N-terminal end. The enzyme was isolated in the supernatant after lysis of the cells and fractionated by ultracentrifugation. Western blotting using anti-histidine antibody showed the presence of a band that migrated at about 47.5 kD on both reducing and non-reducing SDS-PAGE, indicating a monomeric enzyme. Among the carbohydrate acceptors tested, N-acetylneuraminic acid and the gangliosides GD3 and GQ1b were preferred substrates. The cell-free enzyme reaction products obtained were characterized by NMR and mass spectrometry, which indicated the presence of both alpha 2,9- and alpha 2,8-linked polysialyl structure. The K92 neuS gene was used to transform the K1 strain of E. coli, the capsule of which contains only (-8-NeuAc alpha 2-) linkages. Analysis of the polysaccharides isolated from these transformed cells is consistent with the presence of both (-8-NeuAc alpha 2-) and (-9-NeuAc alpha 2-) linkages. It is disclosed that the neuS gene product of E. coli K92 catalyzes the synthesis of polysialic acid with alpha 2,9- and alpha 2,8- linkages in vitro and in vivo. |
| priorityDate | 1998-08-10-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
Total number of triples: 41.