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Production of a soluble and functional recombinant apolipoproteinD in the Pichia pastoris expression system

Armanmehr, S. and Kalhor, H.R. and Tabarraei, A. (2016) Production of a soluble and functional recombinant apolipoproteinD in the Pichia pastoris expression system. Protein Expression and Purification, 121. pp. 157-162.

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Abstract

ApolipoproteinD (ApoD) is a human glycoprotein from the lipocalin family. ApoD contains a conserved central motif of an 8-stranded antiparallel β-sheet, which forms a beta-barrel that can be used for transport and storage of diverse hydrophobic ligands. Due to hydrophobic nature of ApoD, it has been difficult to generate a recombinant version of this protein. In the present work, we aimed at the production of ApoD in the robust Pichia pastoris expression system. To this end, the ApoD gene sequence was synthesized and subcloned for expression in the yeast host cells. Following integration of the ApoD gene into the yeast genomic region using homologous recombination, the ApoD recombinant protein was induced using methanol, reaching its maximum induction at 96 h. Having purified the ApoD recombinant protein by affinity chromatography, we measured the dissociation constant (KD) using its natural ligands: progesterone and arachidonic acid. Our results provide a viable solution to the production of recombinant ApoD protein in lieu of previous obstacles in generating soluble and functional ApoD protein. © 2016 Elsevier Inc. All rights reserved.

Item Type: Article
Additional Information: cited By 0
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 13 Sep 2016 04:42
Last Modified: 13 Sep 2016 08:12
URI: http://eprints.goums.ac.ir/id/eprint/4832

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