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Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides
Molinate is a recalcitrant thiocarbamate used to control grass weeds in rice fields. The recently described molinate hydrolase, from Gulosibacter molinativorax ON4(T), plays a key role in the only known molinate degradation pathway ending in the formation of innocuous compounds. Here we report the c...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4407906/ https://www.ncbi.nlm.nih.gov/pubmed/25905461 http://dx.doi.org/10.1371/journal.pone.0123430 |
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author | Leite, José P. Duarte, Márcia Paiva, Ana M. Ferreira-da-Silva, Frederico Matias, Pedro M. Nunes, Olga C. Gales, Luís |
author_facet | Leite, José P. Duarte, Márcia Paiva, Ana M. Ferreira-da-Silva, Frederico Matias, Pedro M. Nunes, Olga C. Gales, Luís |
author_sort | Leite, José P. |
collection | PubMed |
description | Molinate is a recalcitrant thiocarbamate used to control grass weeds in rice fields. The recently described molinate hydrolase, from Gulosibacter molinativorax ON4(T), plays a key role in the only known molinate degradation pathway ending in the formation of innocuous compounds. Here we report the crystal structure of recombinant molinate hydrolase at 2.27 Å. The structure reveals a homotetramer with a single mononuclear metal-dependent active site per monomer. The active site architecture shows similarities with other amidohydrolases and enables us to propose a general acid-base catalysis mechanism for molinate hydrolysis. Molinate hydrolase is unable to degrade bulkier thiocarbamate pesticides such as thiobencarb which is used mostly in rice crops. Using a structural-based approach, we were able to generate a mutant (Arg187Ala) that efficiently degrades thiobencarb. The engineered enzyme is suitable for the development of a broader thiocarbamate bioremediation system. |
format | Online Article Text |
id | pubmed-4407906 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-44079062015-05-04 Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides Leite, José P. Duarte, Márcia Paiva, Ana M. Ferreira-da-Silva, Frederico Matias, Pedro M. Nunes, Olga C. Gales, Luís PLoS One Research Article Molinate is a recalcitrant thiocarbamate used to control grass weeds in rice fields. The recently described molinate hydrolase, from Gulosibacter molinativorax ON4(T), plays a key role in the only known molinate degradation pathway ending in the formation of innocuous compounds. Here we report the crystal structure of recombinant molinate hydrolase at 2.27 Å. The structure reveals a homotetramer with a single mononuclear metal-dependent active site per monomer. The active site architecture shows similarities with other amidohydrolases and enables us to propose a general acid-base catalysis mechanism for molinate hydrolysis. Molinate hydrolase is unable to degrade bulkier thiocarbamate pesticides such as thiobencarb which is used mostly in rice crops. Using a structural-based approach, we were able to generate a mutant (Arg187Ala) that efficiently degrades thiobencarb. The engineered enzyme is suitable for the development of a broader thiocarbamate bioremediation system. Public Library of Science 2015-04-23 /pmc/articles/PMC4407906/ /pubmed/25905461 http://dx.doi.org/10.1371/journal.pone.0123430 Text en © 2015 Leite et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Leite, José P. Duarte, Márcia Paiva, Ana M. Ferreira-da-Silva, Frederico Matias, Pedro M. Nunes, Olga C. Gales, Luís Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title | Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title_full | Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title_fullStr | Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title_full_unstemmed | Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title_short | Structure-Guided Engineering of Molinate Hydrolase for the Degradation of Thiocarbamate Pesticides |
title_sort | structure-guided engineering of molinate hydrolase for the degradation of thiocarbamate pesticides |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4407906/ https://www.ncbi.nlm.nih.gov/pubmed/25905461 http://dx.doi.org/10.1371/journal.pone.0123430 |
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