高電界パルス誘導自己消化による酵母内分子の非破壊抽出

カテゴリ: 論文誌(論文単位)

グループ名: 【A】基礎・材料・共通部門

発行日: 2021/11/01

タイトル(英語): Non-distractive Extraction of Intracellular Molecules from Yeast using PEF-assisted Autolysis

著者名: 岡本　修治（熊本大学大学院自然科学教育部），村上　鷹児（熊本大学大学院自然科学教育部），卜部　玄（熊本大学大学院自然科学教育部），勝木　淳（熊本大学産業ナノマテリアル研究所）

著者名(英語): Shuji Okamoto (Graduate School of Science & Technology, Kumamoto University), Yoji Murakami (Graduate School of Science & Technology, Kumamoto University), Gen Urabe (Graduate School of Science & Technology, Kumamoto University), Sunao Katsuki (Institute

キーワード: 高電界パルス，非破壊抽出，酵母，自己消化，Native-PAGE　　pulsed electric field，non-destructive extraction，yeast，autolysis，Native-PAGE

要約(英語): This paper discusses the possibility of non-destructive extraction of intracellular proteins of yeast (Saccharomyces cerevisiae) by means of pulsed electric fields (PEFs) combined with autolysis. First, the effects of PEFs (20 kV/cm, 600 ns) on yeast and the subsequent cell wall collapse phenomenon were studied by morphological observation of fluorescence staining. Morphological observation shows that the PEFs increase the permeability of the cell membrane and also disrupt the intracellular vacuoles, whereas the cell wall morphology remains unchanged. However, the cell wall was collapse during the post-PEF incubation. Next, the amount of extracted protein was evaluated by using the BCA assay. The amount of extracted protein in the PEF treated suspension was only a little, whereas the additional incubation promotes the extraction. Lastly, the size of the extracted proteins was investigated by using both Native-PAGE and SDS-PAGE. The PEF-treated and incubated suspension contains the large protein over 250 kDa.

本誌: 電気学会論文誌A（基礎・材料・共通部門誌） Vol.141 No.11 （2021）

本誌掲載ページ: 615-621 p

原稿種別: 論文／日本語

電子版へのリンク: https://www.jstage.jst.go.jp/article/ieejfms/141/11/141_615/_article/-char/ja/