Repository of Research and Investigative Information

Repository of Research and Investigative Information

دانشگاه علوم پزشکی و خدمات بهداشتی درمانی زنجان

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

(2013) Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in pichia pastoris. Electronic Journal of Biotechnology.

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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 (alpha-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.

Item Type: Article
Keywords: Author Keywords:human alpha 1-antitrypsin; Pichia pastoris; recombinant proteins KeyWords Plus:GENE COPY NUMBER; HETEROLOGOUS PROTEIN EXPRESSION; TIME QUANTITATIVE PCR; B SURFACE-ANTIGEN; CODON USAGE BIAS; METHYLOTROPHIC YEAST; SACCHAROMYCES-CEREVISIAE; ANTITRYPSIN DEFICIENCY; CULTURE-CONDITIONS; MOLECULAR-CLONING
Journal or Publication Title: Electronic Journal of Biotechnology
Abstract and Indexing: ISI, Scopus
Quartile : Q3
Volume: 16
Number: 1
Publisher: scopus
Depositing User: خانم مریم زرقانی
URI: http://repository.zums.ac.ir/id/eprint/1737

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