Title of article
Expression and Purification of Functionally Active Recombinant Human Alpha 1-Antitrypsin in Methylotrophic Yeast Pichia Pastoris
Author/Authors
Arjmand، Sareh نويسنده National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran , , Bidram، Elham نويسنده Department of Clinical Biochemistry, Tarbiat Modares University , , Sahebghadam Lotfi، Abbas نويسنده , , Shamsara، Mehdi نويسنده , , Mowla، Seyed Javad نويسنده ,
Issue Information
فصلنامه با شماره پیاپی 10 سال 2011
Pages
8
From page
127
To page
134
Abstract
Human alpha 1-antitrypsin (AAT) cDNA was obtained from HepG2 cell lines. After PCR and construction of expression vector pPICZα-AAT, human AAT was expressed in the yeast Pichia pastoris (P.pastoris) in a secretary manner and under the control of inducible alcohol oxidase 1 (AOX1) promoter. The amount of AAT protein in medium was measured as 60 mg/l 72 hr after induction with methanol. Results indicated the presence of protease inhibitory function of the protein against elastase. Purification was done using His-tag affinity chromatography. Due to the different patterns of glycosylation in yeast and human, the recombinant AAT showed different SDS-PAGE patterns compared to that of serum-derived AAT while pI shifted from 4.9 in native AAT compared to 5.2 in recombinant AAT constructed in this study.
Journal title
AJMB Avicenna Journal of Medical Biotechnology
Serial Year
2011
Journal title
AJMB Avicenna Journal of Medical Biotechnology
Record number
1982821
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