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A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove
The use of culture-independent approaches, such as metagenomics, provides complementary access to environmental microbial diversity. Mangrove environments represent a highly complex system with plenty of opportunities for finding singular functions. In this study we performed a functional screening...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615308/ https://www.ncbi.nlm.nih.gov/pubmed/28952541 http://dx.doi.org/10.3390/bioengineering4030062 |
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author | Soares, Fábio Lino Marcon, Joelma Pereira e Silva, Michele de Cássia Khakhum, Nittaya Cerdeira, Louise Teixeira Ottoni, Júlia Ronzella Domingos, Daniela Ferreira Taketani, Rodrigo Gouvea de Oliveira, Valéria Maia Lima, André Oliveira de Souza Azevedo, João Lucio Rodrigues, Jorge Luiz Mazza Andreote, Fernando Dini |
author_facet | Soares, Fábio Lino Marcon, Joelma Pereira e Silva, Michele de Cássia Khakhum, Nittaya Cerdeira, Louise Teixeira Ottoni, Júlia Ronzella Domingos, Daniela Ferreira Taketani, Rodrigo Gouvea de Oliveira, Valéria Maia Lima, André Oliveira de Souza Azevedo, João Lucio Rodrigues, Jorge Luiz Mazza Andreote, Fernando Dini |
author_sort | Soares, Fábio Lino |
collection | PubMed |
description | The use of culture-independent approaches, such as metagenomics, provides complementary access to environmental microbial diversity. Mangrove environments represent a highly complex system with plenty of opportunities for finding singular functions. In this study we performed a functional screening of fosmid libraries obtained from an oil contaminated mangrove site, with the purpose of identifying clones expressing hydrolytic activities. A novel gene coding for a β-N-acetylhexosaminidase with 355 amino acids and 43KDa was retrieved and characterized. The translated sequence showed only 38% similarity to a β-N-acetylhexosaminidase gene in the genome of Veillonella sp. CAG:933, suggesting that it might constitute a novel enzyme. The enzyme was expressed, purified, and characterized for its enzymatic activity on carboxymethyl cellulose, p-Nitrophenyl-2acetamide-2deoxy-β-d-glucopyranoside, p-Nitrophenyl-2acetamide-2deoxy-β-d-galactopyranoside, and 4-Nitrophenyl β-d-glucopyranoside, presenting β-N-acetylglucosaminidase, β-glucosidase, and β-1,4-endoglucanase activities. The enzyme showed optimum activity at 30 °C and pH 5.5. The characterization of the putative novel β-N-acetylglucosaminidase enzyme reflects similarities to characteristics of the environment explored, which differs from milder conditions environments. This work exemplifies the application of cultivation-independent molecular techniques to the mangrove microbiome for obtaining a novel biotechnological product. |
format | Online Article Text |
id | pubmed-5615308 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-56153082017-09-28 A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove Soares, Fábio Lino Marcon, Joelma Pereira e Silva, Michele de Cássia Khakhum, Nittaya Cerdeira, Louise Teixeira Ottoni, Júlia Ronzella Domingos, Daniela Ferreira Taketani, Rodrigo Gouvea de Oliveira, Valéria Maia Lima, André Oliveira de Souza Azevedo, João Lucio Rodrigues, Jorge Luiz Mazza Andreote, Fernando Dini Bioengineering (Basel) Article The use of culture-independent approaches, such as metagenomics, provides complementary access to environmental microbial diversity. Mangrove environments represent a highly complex system with plenty of opportunities for finding singular functions. In this study we performed a functional screening of fosmid libraries obtained from an oil contaminated mangrove site, with the purpose of identifying clones expressing hydrolytic activities. A novel gene coding for a β-N-acetylhexosaminidase with 355 amino acids and 43KDa was retrieved and characterized. The translated sequence showed only 38% similarity to a β-N-acetylhexosaminidase gene in the genome of Veillonella sp. CAG:933, suggesting that it might constitute a novel enzyme. The enzyme was expressed, purified, and characterized for its enzymatic activity on carboxymethyl cellulose, p-Nitrophenyl-2acetamide-2deoxy-β-d-glucopyranoside, p-Nitrophenyl-2acetamide-2deoxy-β-d-galactopyranoside, and 4-Nitrophenyl β-d-glucopyranoside, presenting β-N-acetylglucosaminidase, β-glucosidase, and β-1,4-endoglucanase activities. The enzyme showed optimum activity at 30 °C and pH 5.5. The characterization of the putative novel β-N-acetylglucosaminidase enzyme reflects similarities to characteristics of the environment explored, which differs from milder conditions environments. This work exemplifies the application of cultivation-independent molecular techniques to the mangrove microbiome for obtaining a novel biotechnological product. MDPI 2017-07-09 /pmc/articles/PMC5615308/ /pubmed/28952541 http://dx.doi.org/10.3390/bioengineering4030062 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Soares, Fábio Lino Marcon, Joelma Pereira e Silva, Michele de Cássia Khakhum, Nittaya Cerdeira, Louise Teixeira Ottoni, Júlia Ronzella Domingos, Daniela Ferreira Taketani, Rodrigo Gouvea de Oliveira, Valéria Maia Lima, André Oliveira de Souza Azevedo, João Lucio Rodrigues, Jorge Luiz Mazza Andreote, Fernando Dini A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title | A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title_full | A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title_fullStr | A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title_full_unstemmed | A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title_short | A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove |
title_sort | novel multifunctional β-n-acetylhexosaminidase revealed through metagenomics of an oil-spilled mangrove |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615308/ https://www.ncbi.nlm.nih.gov/pubmed/28952541 http://dx.doi.org/10.3390/bioengineering4030062 |
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