Nucleic acid sequences and expression systems for heparinase II and heparinase III derived from Flavobacterium heparinum
US5681733A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jun 10, 1994 |
| Grant date | Oct 28, 1997 |
| Priority date | — |
| Expiry date | Jun 10, 2014 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y402/02007
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.