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タイトル: NMR Analysis on Molecular Interaction of Lignin with Amino Acid Residues of Carbohydrate-Binding Module from Trichoderma reesei Cel7A
著者: Tokunaga, Yuki
Nagata, Takashi  kyouindb  KAKEN_id
Suetomi, Takashi
Oshiro, Satoshi
Kondo, Keiko
Katahira, Masato
Watanabe, Takashi
著者名の別形: 徳永, 有希
永田, 崇
大城, 理志
近藤, 敬子
片平, 正人
渡邊, 隆司
キーワード: Hydrolases
Solution-state NMR
発行日: 13-Feb-2019
出版者: Springer Nature
誌名: Scientific Reports
巻: 9
論文番号: 1977
抄録: Lignocellulosic biomass is anticipated to serve as a platform for green chemicals and fuels. Nonproductive binding of lignin to cellulolytic enzymes should be avoided for conversion of lignocellulose through enzymatic saccharification. Although carbohydrate-binding modules (CBMs) of cellulolytic enzymes strongly bind to lignin, the adsorption mechanism at molecular level is still unclear. Here, we report NMR-based analyses of binding sites on CBM1 of cellobiohydrolase I (Cel7A) from a hyper-cellulase-producing fungus, Trichoderma reesei, with cellohexaose and lignins from Japanese cedar (C-MWL) and Eucalyptus globulus (E-MWL). A method was established to obtain properly folded TrCBM1. Only TrCBM1 that was expressed in freshly transformed E. coli had intact conformation. Chemical shift perturbation analyses revealed that TrCBM1 adsorbed cellohexaose in highly specific manner via two subsites, flat plane surface and cleft, which were located on the opposite side of the protein surface. Importantly, MWLs were adsorbed at multiple binding sites, including the subsites, having higher affinity than cellohexaose. G6 and Q7 were involved in lignin binding on the flat plane surface of TrCBM1, while cellohexaose preferentially interacted with N29 and Q34. TrCBM1 used much larger surface area to bind with C-MWL than E-MWL, indicating the mechanisms of adsorption toward hardwood and softwood lignins are different.
記述: セルラーゼとリグニンの相互作用をはじめて分子レベルで包括的に解明 --バイオマス変換や酵素科学に貢献--. 京都大学プレスリリース. 2019-02-14.
著作権等: © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
URI: http://hdl.handle.net/2433/236471
DOI(出版社版): 10.1038/s41598-018-38410-9
関連リンク: http://www.kyoto-u.ac.jp/ja/research/research_results/2018/190213_1.html
出現コレクション:学術雑誌掲載論文等

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