کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10843331 | 1069219 | 2010 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression and characterization of a Grifola frondosa hydrophobin in Pichia pastoris
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Hydrophobins are small secreted proteins produced by filamentous fungi. Being amphipathic and self-assembling, hydrophobins have drawn great attention since their discovery. The increase of production can reduce the cost and open up several new applications of hydrophobins. We successfully expressed recombinant Class I hydrophobin HGFI (rHGFI) by using pPIC9 vector with an alcohol oxidase 1 promoter in Pichia pastoris. Tricine-SDS-PAGE and Western blotting demonstrated that rHGFI, an 8Â kDa protein, was secreted into the culture medium. The culture conditions of the transformant strain were optimized by controlling the methanol concentration and induction time. Ultrafiltration and reverse-phase high performance liquid chromatography were used to perform a large-scale purification of rHGFI. A stable production of rHGFI around 86Â mg/L was achieved after the two-step purification. X-ray photoelectron spectroscopy and water contact angle measurements indicated that the functional rHGFI could self-assemble on hydrophobic siliconized glass and Teflon as well as on hydrophilic mica surfaces. A methylthiazol tetrazolium assay showed that rHGFI film could facilitate human aortic smooth muscle cell proliferation due to its cytocompatibility.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 72, Issue 1, July 2010, Pages 19-25
Journal: Protein Expression and Purification - Volume 72, Issue 1, July 2010, Pages 19-25
نویسندگان
Zefang Wang, Shuren Feng, Yujian Huang, Shan Li, Haijin Xu, Xiuming Zhang, Yanling Bai, Mingqiang Qiao,