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Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment
Enzymatic breakdown is an attractive cellulose utilisation method with a low environmental load. Its high temperature operation could promote saccharification and lower contamination risk. Here we report a hyper-thermostable cellobiohydrolase (CBH), named HmCel6A and its variant HmCel6A-3SNP that we...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8940973/ https://www.ncbi.nlm.nih.gov/pubmed/35318423 http://dx.doi.org/10.1038/s42003-022-03195-1 |
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author | Takeda, Migiwa Baba, Seiki Okuma, Jiro Hirose, Yoshitsugu Nishimura, Asuka Takata, Masaki Oda, Kohei Shibata, Daisuke Kumasaka, Takashi Kondo, Yasuhiro |
author_facet | Takeda, Migiwa Baba, Seiki Okuma, Jiro Hirose, Yoshitsugu Nishimura, Asuka Takata, Masaki Oda, Kohei Shibata, Daisuke Kumasaka, Takashi Kondo, Yasuhiro |
author_sort | Takeda, Migiwa |
collection | PubMed |
description | Enzymatic breakdown is an attractive cellulose utilisation method with a low environmental load. Its high temperature operation could promote saccharification and lower contamination risk. Here we report a hyper-thermostable cellobiohydrolase (CBH), named HmCel6A and its variant HmCel6A-3SNP that were isolated metagenomically from hot spring sediments and expressed in Escherichia coli. They are classified into glycoside hydrolases family 6 (GH6). HmCel6A-3SNP had three amino acid replacements to HmCel6A (P88S/L230F/F414S) and the optimum temperature at 95 °C, while HmCel6A did it at 75 °C. Crystal structure showed conserved features among GH6, a (β/α)(8)-barrel core and catalytic residues, and resembles TfCel6B, a bacterial CBH II of Thermobifida fusca, that had optimum temperature at 60 °C. From structure-function studies, we discuss unique structural features that allow the enzyme to reach its high thermostability level, such as abundance of hydrophobic and charge-charge interactions, characteristic metal bindings and disulphide bonds. Moreover, structure and surface plasmon resonance analysis with oligosaccharides suggested that the contribution of an additional tryptophan located at the tunnel entrance could aid in substrate recognition and thermostability. These results may help to design efficient enzymes and saccharification methods for cellulose working at high temperatures. |
format | Online Article Text |
id | pubmed-8940973 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-89409732022-04-08 Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment Takeda, Migiwa Baba, Seiki Okuma, Jiro Hirose, Yoshitsugu Nishimura, Asuka Takata, Masaki Oda, Kohei Shibata, Daisuke Kumasaka, Takashi Kondo, Yasuhiro Commun Biol Article Enzymatic breakdown is an attractive cellulose utilisation method with a low environmental load. Its high temperature operation could promote saccharification and lower contamination risk. Here we report a hyper-thermostable cellobiohydrolase (CBH), named HmCel6A and its variant HmCel6A-3SNP that were isolated metagenomically from hot spring sediments and expressed in Escherichia coli. They are classified into glycoside hydrolases family 6 (GH6). HmCel6A-3SNP had three amino acid replacements to HmCel6A (P88S/L230F/F414S) and the optimum temperature at 95 °C, while HmCel6A did it at 75 °C. Crystal structure showed conserved features among GH6, a (β/α)(8)-barrel core and catalytic residues, and resembles TfCel6B, a bacterial CBH II of Thermobifida fusca, that had optimum temperature at 60 °C. From structure-function studies, we discuss unique structural features that allow the enzyme to reach its high thermostability level, such as abundance of hydrophobic and charge-charge interactions, characteristic metal bindings and disulphide bonds. Moreover, structure and surface plasmon resonance analysis with oligosaccharides suggested that the contribution of an additional tryptophan located at the tunnel entrance could aid in substrate recognition and thermostability. These results may help to design efficient enzymes and saccharification methods for cellulose working at high temperatures. Nature Publishing Group UK 2022-03-22 /pmc/articles/PMC8940973/ /pubmed/35318423 http://dx.doi.org/10.1038/s42003-022-03195-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access 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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Takeda, Migiwa Baba, Seiki Okuma, Jiro Hirose, Yoshitsugu Nishimura, Asuka Takata, Masaki Oda, Kohei Shibata, Daisuke Kumasaka, Takashi Kondo, Yasuhiro Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title | Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title_full | Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title_fullStr | Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title_full_unstemmed | Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title_short | Metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
title_sort | metagenomic mining and structure-function studies of a hyper-thermostable cellobiohydrolase from hot spring sediment |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8940973/ https://www.ncbi.nlm.nih.gov/pubmed/35318423 http://dx.doi.org/10.1038/s42003-022-03195-1 |
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