グライコシンターゼ化による反転型加水分解酵素のグリコシド合成触媒への変換
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概要
- 論文の詳細を見る
還元末端キシロース遊離エキソオリゴキシラナーゼ（Rex, EC. 3.2.1.156）は，糖加水分解酵素ファミリー 8 に属する反転型キシラン分解酵素であり，キシロオリゴ糖から還元末端キシロース（X1）を遊離する反応を触媒する。Rex は X1 の存在下のみ，α キシロビオシルフルオリド（α-X2F）を加水分解する，Hehre-合成-加水分解反応を示した。一般塩基触媒残基（D263）変異酵素ライブラリーを作製し，グライコシンターゼ活性により α-X2F と X1 からキシロトリオース（X3）を最も良く蓄積する D263C 変異酵素を選抜した。しかしながら，D263C のフッ素イオン遊離活性は親酵素より低かった。次に，一般塩基触媒残基と共に求核試薬である水を保持している残基である Y198 をフェニルアラニンに変異させたところ，フッ素イオン遊離活性が低下することなく加水分解活性のみに大幅に低下が見られた。Rex の Y198F 変異酵素は D263C よりも X3 の蓄積が多かった。最近になり，糖加水分解酵素ファミリー 95 に属する α1,2 フコシダーゼのグライコシンターゼ化が報告された。この場合は，一般塩基触媒残基と相互作用している別の残基に変異を入れることが有効であった。両者ともに，一般塩基触媒残基は変異させない方が良かった。
- 2009-01-31
著者
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Kitaoka Motomitsu
National Food Research Institute, National Agriculture and Food Research Organization
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Fushinobu Shinya
Department of Biotechnology, The University of Tokyo
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Katayama Tetsuyuki
Laboratory Of Nutritional Science Faculty Of Education Hiroshima University
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山本 憲二
京大・農・食工
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Katayama Takane
Research Institute For Bioresources And Biotechnology Ishikawa Prefectural University
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Fushinobu Shinya
Dep. Of Biotechnology Graduate School Of Agricultural And Life Sciences The Univ. Of Tokyo
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Fushinobu Shinya
Department Of Biotechnology Graduate School Of Agricultural And Life Sciences The University Of Toky
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Kitaoka M
Enzyme Laboratory National Food Research Institute
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Kitaoka M
Chubu Univ. Aichi Jpn
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Kitaoka Motomitsu
National Food Res. Inst.
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Kitaoka Motomitsu
Enzyme Laboratory National Food Research Institute National Agriculture And Food Research Organizati
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Honda Yuji
National Food Research Institute, National Agriculture and Food Research Organization
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Hidaka Masafumi
National Food Research Institute, National Agriculture and Food Research Organization
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Katayama Takane
Ishikawa Prefectural University
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Yamamoto Kenji
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
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Honda Yuji
National Food Research Institute 2-1-12 Kannondai
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Hidaka Masafumi
Department Of Biotechnology The University Of Tokyo
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Kitaoka Motomitsu
National Food Research Institute National Agriculture And Food Research Organization
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HONDA Yuji
Ishikawa Prefectural University
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Yamamoto Kenji
Division Of Integrated Life Science Graduate School Of Biostudies Kyoto University
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Yokozeki Kenzo
Research Institute For Bioresources And Biotechnology Ishikawa Prefectural University
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Fushinobu Shinya
Department Of Biotechnology Graduate School Of Agricultural And Life Sciences The University Of Tokyo
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Kitaoka Motomitsu
National Agriculture And Food Research Organization National Food Research Institute
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