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Purification, cloning and functional characterization of fructan : Fructan 1-fructosyltransferase from edible burdock (Arctium lappa L..)
http://hdl.handle.net/10659/1768
http://hdl.handle.net/10659/17687e18b407-ff52-439e-8d74-4677e137ac0b
名前 / ファイル | ライセンス | アクション |
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S-810_Ueno.pdf (1.3 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2011-03-31 | |||||
タイトル | ||||||
タイトル | Purification, cloning and functional characterization of fructan : Fructan 1-fructosyltransferase from edible burdock (Arctium lappa L..) | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Abe, Masami
× Abe, Masami× Ueno, Keiji× Ishiguro, Yojiro× Omori, Toshima× Onodera, Shuichi× Shiomi, Norio |
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著者別名 | ||||||
姓名 | 阿部, 雅美 | |||||
著者別名 | ||||||
姓名 | 上野, 敬司 | |||||
著者別名 | ||||||
姓名 | 石黒, 陽二郎 | |||||
著者別名 | ||||||
姓名 | 大森, 俊馬 | |||||
著者別名 | ||||||
姓名 | 小野寺, 秀一 | |||||
著者別名 | ||||||
姓名 | 塩見, 徳夫 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | A fructan:fructan 1-fructosyltransferase (1-FFT) was purified edible for the first time, and a cDNA encoding 1-FFT was isolated from the plant. Two 1-FFTs named 1-FFTa and 1-FFTb were purified from extract of edible burdock by sulfate precipitation, followed by chromatographies on DEAE-Sepharose CL-6B, Toypearl HW-55S and Sehadex G-100 columns. Inulin-type fructan such as nystose and frustooligosaccharides with higher DP were produced from 1-kestose by purified 1-FFTs. The general properties of both purified enzymes were very similar to each other. The purified 1-FFTa and 1-FFTb showed a single band by native PAGE and two bands, relative molecular messes of about 46,000 and 17,500 for 1-FFTa and 46,000 and 17,000 for 1-FFTb, by SDS-PAGE, respectively. The N-terminal sequences of the 46,000 peptides of both enzymes were the same, and those of the 17,000 and the 17,000 peptides were also identical. Based on the sequences, the 1-FFT cDNA named alft1 was cloned. The alft1 encoded a polypeptide of 617 amino acids. The relative molecular mass and pI of the mature protein region of deduced polypeptide were calculated to be 60,213 and 4.89, respectivety. The characteristics of recombinant protein produced by Pichia pastoris closely resembled those of the native 1-FFTs purified from edible burdock. In this study, we demonstrated that alft1 encoding edible burdock 1-FFT was involved in the elongation of fructosyl chains of fructooligosaccharides in edible burdock. ごぼうから初めてフルクタン:フルクタン1-フルクトシルトランスフェラーゼ(1-FFT)を精製し,それをコードするcDNAをクローニングした。1- FFTaおよび1-FFTbと名付けた2種の1-FFTが硫安分画,DEAE-Sepharose CL-6B, Toyopearl HW-55SおよびSephadex G-100の各種クロマトグラフィーを行うことによりごぼう抽出液から精製された。ニストースや高い重合度をもつフルクトオリゴ糖などのイヌリン型フルクタンが1-ケストースから精製1-FFTにより生成された。両精製酵素の一般性質はよく似ていた.1-FFTaおよび1-FFTb両精製酵素はネイティブ PAGEにより単一バンドを示したが,SDS-PAGEでは1-FFTaでは分子量が約46,000と17,500の位置に,1-FFTbでは約 46,000と17,000の位置に,それぞれ2本のバンドを示した。両酵素の46,000のペプチドのN末端配列は同じ配列であり,両酵素の 17,500と17,000のペプチドのN末端配列もまた同じ配列であった。これらの配列に基づいてalft1と名付けた1-FFT cDNAをクローニングした。alft1は617アミノ酸からなるポリペプチドをコードしていた。推定されるアミノ酸配列の成熟タンパク質領域の分子量と等電点は60,213および4.89と計算された。Pichia pastorisで生産された組み換えタンパク質の性質はごぼうから精製した1-FFTとよく似ていた。本研究において,われわれはごぼう由来1-FFTとそれをコードするalft1がごぼうのフルクトオリゴ糖のフルクトシル鎖の伸長に関与することを示した。 | |||||
書誌情報 |
Journal of applied glycoscience 巻 56, 号 4, p. 239-246, 発行日 2009-11 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1344-7882 | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
その他のタイトル | ||||||
その他のタイトル | ごぼう (Arctium lappa L.) 由来フルクタン : フルクタン1-フルクトシルトランスフェラーゼの精製,クローニングと機能解析 | |||||
出版者 | ||||||
出版者 | 日本応用糖質科学会 | |||||
資源タイプ | ||||||
内容記述タイプ | Other | |||||
内容記述 | Article |