Method of using aminopeptidase to produce cleaved polypeptides
US7632658B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Apr 27, 2007 |
| Grant date | Dec 15, 2009 |
| Priority date | — |
| Expiry date | Feb 11, 2028 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12R2001/19
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A gram-negative bacterial cell is described that is deficient in a chromosomal gene present in a wild-type such cell which gene shares at least 80% sequence identity with the native sequence of the yfcK gene and encodes an aminopeptidase. Alternatively, a gram-negative bacterial cell is deficient in a chromosomal gene present in a wild-type such cell which gene encodes an aminopeptidase that shares at least 80% sequence identity with the native sequence of aminopeptidase b2324. Either of these types of cells, when comprising a nucleic acid encoding a heterologous polypeptide, produces an N-terminal unclipped polypeptide when it is cultured and the polypeptide recovered, with virtually no N-terminal clipped polypeptide produced as an impurity. Conversely, a method is provided for cleaving an N-terminal amino acid from a polypeptide comprising contacting the polypeptide with an aminopeptidase sharing at least 80% sequence identity with the native sequence of aminopeptidase b2324.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.