Process for the enzymatic preparation of 7-aminocephalosporanic acid
US5354667A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Aug 5, 1991 |
| Grant date | Oct 11, 1994 |
| Priority date | — |
| Expiry date | Aug 5, 2011 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12P35/02
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A process for the production of an E. coli strain producing high yields of 7.beta.-(4-carboxybutanamido) cephalosporin acylase, comprises: (a) digesting the DNA of a microorganism whose DNA includes the sequence encoding 7.beta.-(4-carboxybutanamido) cephalosporin acylase, and forming a plasmidic library; (b) transforming, with the sequences in the plasmidic library, an auxotrophic E. coli host; (c) selecting for transformed E. coli hosts containing the acylase sequence by growth on a suitable medium; (d) isolating the vector containing the acylase sequence, digesting the vector, and ligating the DNA sequences obtained into an E. coli vector under control of an E. coli promoter; (e) repeating selection procedure of step (c) ; (f) using the vectors from the selected E. coli hosts to transform an E. coli host lacking substantial .beta.-lactamase activity. 7-Aminocephalosporanic acid and its derivatives can be prepared by reaction of substrates like 7.beta.-(4-carboxybutanamido) cephalosporanic acid with 7.beta.-(4-carboxybutanamido) cephalosporin acylase produced by the E. coli strain produced by the above process.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.