論文

査読有り
2015年9月1日

A method of gentle hydration to prepare oil-free giant unilamellar vesicles that can confine enzymatic reactions

Biochemistry and Biophysics Reports
  • K. Shohda
  • ,
  • K. Takahashi
  • ,
  • A. Suyama

3
開始ページ
76
終了ページ
82
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.bbrep.2015.07.005
出版者・発行元
Elsevier

We report a new and improved method to prepare, by gentle hydration of lipid films, oil-free giant unilamellar vesicles (GUVs), in which enzymatic reactions can be encapsulated. The traditional method of gentle hydration requires very low concentrations of metal ions, whereas enzymatic reactions generally require mono- and divalent metal ions at physiological concentrations. In order to improve the production of oil-free GUVs that can confine enzymatic reactions, we developed a novel method also based on gentle hydration, but in which the precursor lipid film was doped with both 1,2-dioleoyl-. sn-glycero-3-phosphoethanolamine-. N-[methoxy(polyethylene glycol)-2000] (PEGylated lipid) and sugar. Close examination of the size, shape, and lamellarity of vesicles prepared in this manner demonstrated that the process improves the production of oil-free GUVs even at low temperatures and physiological salt concentrations. PEGylated lipid and sugar were found to synergistically improve GUV formation. Finally, we demonstrate the successful enzymatic synthesis of RNA within oil-free GUVs that were prepared on ice.

リンク情報
DOI
https://doi.org/10.1016/j.bbrep.2015.07.005
ID情報
  • DOI : 10.1016/j.bbrep.2015.07.005
  • ISSN : 2405-5808
  • SCOPUS ID : 84938779062

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