| Revista: | Brazilian archives of biology and technology |
| Base de datos: | PERIÓDICA |
| Número de sistema: | 000366259 |
| ISSN: | 1516-8913 |
| Autors: | Liu, Cheng-Cheng1 Zhai, Le1 Shi, Ying1 Yang, Ke-Wu1 |
| Institucions: | 1Northwest University, College of Chemistry and Materials Science, Xi'an, Shaanxi. China |
| Any: | 2013 |
| Període: | Sep-Oct |
| Volum: | 56 |
| Número: | 5 |
| Paginació: | 811-816 |
| País: | Brasil |
| Idioma: | Inglés |
| Tipo de documento: | Artículo |
| Enfoque: | Experimental, analítico |
| Resumen en inglés | A simple and effective salting-out method was developed for the purification of the metallo-β-lactamase CcrA from Bacteroides fragilis based on the plasmid pMSZ02, in which the crude protein secreted into growth medium was precipitated by 80% sulfate saturation of the medium and purified with Q-Sepharose to offer pure CcrA with yield of 20.1 mg per litter medium. The dependence of the amount of protein precipitation on sulfate saturation was investigated, which showed that more than 80% sulfate saturation resulted the maximum protein precipitated. The purified CcrA was evaluated by steady-state kinetics using penicillin G and cephalothin V as substrates, which showed the Kmvalues of 68±2 and 17±2 µM and Kcatvalues of 63±1 and 102±3 S-1, respectively. The comparison with the data of the protein from literature method showed that the salting-out method was viable, and it could be useful for the purification of other proteins secreted into growth medium |
| Disciplines | Biología, Química |
| Paraules clau: | Bacterias, Bioquímica, Microbiología, Bacteroides fragilis, Metalo-beta-lactamasa, Purificación, Precipitación salina |
| Keyword: | Biology, Chemistry, Bacteria, Biochemistry, Microbiology, Bacteroides fragilis, Metallo-betha-lactamase, Purification, Salting out process |
| Text complet: | Texto completo (Ver HTML) |
