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Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps
Cellobiohydrolase 1 from Trichoderma reesei (TrCel7A) processively hydrolyses cellulose into cellobiose. Although enzymatic techniques have been established as promising tools in biofuel production, a clear understanding of the motor's mechanistic action has yet to be revealed. Here, we develop...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682103/ https://www.ncbi.nlm.nih.gov/pubmed/26657780 http://dx.doi.org/10.1038/ncomms10149 |
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author | Brady, Sonia K. Sreelatha, Sarangapani Feng, Yinnian Chundawat, Shishir P. S. Lang, Matthew J |
author_facet | Brady, Sonia K. Sreelatha, Sarangapani Feng, Yinnian Chundawat, Shishir P. S. Lang, Matthew J |
author_sort | Brady, Sonia K. |
collection | PubMed |
description | Cellobiohydrolase 1 from Trichoderma reesei (TrCel7A) processively hydrolyses cellulose into cellobiose. Although enzymatic techniques have been established as promising tools in biofuel production, a clear understanding of the motor's mechanistic action has yet to be revealed. Here, we develop an optical tweezers-based single-molecule (SM) motility assay for precision tracking of TrCel7A. Direct observation of motility during degradation reveals processive runs and distinct steps on the scale of 1 nm. Our studies suggest TrCel7A is not mechanically limited, can work against 20 pN loads and speeds up when assisted. Temperature-dependent kinetic studies establish the energy requirements for the fundamental stepping cycle, which likely includes energy from glycosidic bonds and other sources. Through SM measurements of isolated TrCel7A domains, we determine that the catalytic domain alone is sufficient for processive motion, providing insight into TrCel7A's molecular motility mechanism. |
format | Online Article Text |
id | pubmed-4682103 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46821032015-12-29 Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps Brady, Sonia K. Sreelatha, Sarangapani Feng, Yinnian Chundawat, Shishir P. S. Lang, Matthew J Nat Commun Article Cellobiohydrolase 1 from Trichoderma reesei (TrCel7A) processively hydrolyses cellulose into cellobiose. Although enzymatic techniques have been established as promising tools in biofuel production, a clear understanding of the motor's mechanistic action has yet to be revealed. Here, we develop an optical tweezers-based single-molecule (SM) motility assay for precision tracking of TrCel7A. Direct observation of motility during degradation reveals processive runs and distinct steps on the scale of 1 nm. Our studies suggest TrCel7A is not mechanically limited, can work against 20 pN loads and speeds up when assisted. Temperature-dependent kinetic studies establish the energy requirements for the fundamental stepping cycle, which likely includes energy from glycosidic bonds and other sources. Through SM measurements of isolated TrCel7A domains, we determine that the catalytic domain alone is sufficient for processive motion, providing insight into TrCel7A's molecular motility mechanism. Nature Publishing Group 2015-12-10 /pmc/articles/PMC4682103/ /pubmed/26657780 http://dx.doi.org/10.1038/ncomms10149 Text en Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Brady, Sonia K. Sreelatha, Sarangapani Feng, Yinnian Chundawat, Shishir P. S. Lang, Matthew J Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title | Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title_full | Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title_fullStr | Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title_full_unstemmed | Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title_short | Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps |
title_sort | cellobiohydrolase 1 from trichoderma reesei degrades cellulose in single cellobiose steps |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682103/ https://www.ncbi.nlm.nih.gov/pubmed/26657780 http://dx.doi.org/10.1038/ncomms10149 |
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