Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastoris



Document title: Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastoris
Journal: Electronic journal of biotechnology
Database: PERIÓDICA
System number: 000357887
ISSN: 0717-3458
Authors: 1
2
2
1
Institutions: 1Tarbiat Modares University, Faculty of Biological Sciences, Teherán. Irán
2National Institute of Genetic Engineering and Biotechnology, Shahrak-e Pajoohesh, Teherán. Irán
Year:
Volumen: 16
Number: 1
Country: Chile
Language: Inglés
Document type: Artículo
Approach: Experimental, aplicado
English abstract 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
Disciplines: Biología
Keyword: Biotecnología,
Bioproducción,
Antitripsina-alfa 1,
Proteínas recombinantes,
Pichia pastoris
Keyword: Biology,
Biotechnology,
Bioproduction,
Antitrypsin-alpha 1,
Recombinant proteins,
Pichia pastoris
Full text: Texto completo (Ver HTML)