کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020220 1542320 2016 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Production of a soluble and functional recombinant apolipoproteinD in the Pichia pastoris expression system
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Production of a soluble and functional recombinant apolipoproteinD in the Pichia pastoris expression system
چکیده انگلیسی


• Expression of human ApoD protein in the Pichia pastoris expression system as a secreted protein.
• Production of functional and soluble form of ApoD without introducing any mutation.
• High yield of purified ApoD using one step purification.

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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 121, May 2016, Pages 157–162
نویسندگان
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