| Revista: | Electronic journal of biotechnology |
| Base de datos: | PERIÓDICA |
| Número de sistema: | 000357887 |
| ISSN: | 0717-3458 |
| Autores: | Arjmand, Sareh1 Lotfi, Abbas Sahebghadam2 Shamsara, Mehdi2 Mowla, Seyed Javad1 |
| Instituciones: | 1Tarbiat Modares University, Faculty of Biological Sciences, Teherán. Irán 2National Institute of Genetic Engineering and Biotechnology, Shahrak-e Pajoohesh, Teherán. Irán |
| Año: | 2013 |
| Volumen: | 16 |
| Número: | 1 |
| País: | Chile |
| Idioma: | Inglés |
| Tipo de documento: | Artículo |
| Enfoque: | Experimental, aplicado |
| Resumen en inglés | Background: Human alpha 1-antitrypsin (AAT) is a potent inhibitor of multiple serine proteases, and protects tissues against their harmful effects. Individuals with reduced or abnormal production of this inhibitor need intravenous administration of exogenous protein. In this study, we employed the methylotrophic (methanol utilizing) yeast Pichia pastoris (P. pastoris) as a preferential host for efficient production and secretion of recombinant AAT. Furthermore, we examined different strategies to maximize the yield of the secreted protein. Results: Our findings revealed that optimizing the codon usage of AAT gene for P. pastoris had positive effects on the level of secreted AAT under the control of inducible alcohol oxidase 1 (AOX1) and constitutive glycerol aldehyde phosphate dehydrogenase (GAP) promoters. Compared to AOX1, the GAP promoter increased the yield of AAT by more than two fold. It was also demonstrated that the human AAT native signal sequence was more effective than the well-known yeast signal sequence, alpha mating factor (α-MF). Doubling gene dosage nearly doubled the production of AAT, though dosages exceeding this limit had negative effects on the yield. Conclusion: P. pastoris is shown to be an efficient expression system for production of recombinant and biologically active AAT. Also different strategies could be used to elevate the amount of this secretable protein |
| Disciplinas: | Biología |
| Palabras clave: | Biotecnología, Bioproducción, Antitripsina-alfa 1, Proteínas recombinantes, Pichia pastoris |
| Keyword: | Biology, Biotechnology, Bioproduction, Antitrypsin-alpha 1, Recombinant proteins, Pichia pastoris |
| Texto completo: | Texto completo (Ver HTML) |
