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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
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.
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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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