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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| Format: | Article |
| Language: | English |
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MDPI AG
2017-07-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/4/3/62 |
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| author | Fábio Lino Soares Joelma Marcon Michele de Cássia Pereira e Silva Nittaya Khakhum Louise Teixeira Cerdeira Júlia Ronzella Ottoni Daniela Ferreira Domingos Rodrigo Gouvea Taketani Valéria Maia de Oliveira André Oliveira de Souza Lima João Lucio Azevedo Jorge Luiz Mazza Rodrigues Fernando Dini Andreote |
| author_facet | Fábio Lino Soares Joelma Marcon Michele de Cássia Pereira e Silva Nittaya Khakhum Louise Teixeira Cerdeira Júlia Ronzella Ottoni Daniela Ferreira Domingos Rodrigo Gouvea Taketani Valéria Maia de Oliveira André Oliveira de Souza Lima João Lucio Azevedo Jorge Luiz Mazza Rodrigues Fernando Dini Andreote |
| author_sort | Fábio Lino Soares |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-12T06:47:22Z |
| format | Article |
| id | doaj.art-91ba5dab2f3a477b961829d6749a38f1 |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-12T06:47:22Z |
| publishDate | 2017-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-91ba5dab2f3a477b961829d6749a38f12023-09-03T00:30:15ZengMDPI AGBioengineering2306-53542017-07-01436210.3390/bioengineering4030062bioengineering4030062A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled MangroveFábio Lino Soares0Joelma Marcon1Michele de Cássia Pereira e Silva2Nittaya Khakhum3Louise Teixeira Cerdeira4Júlia Ronzella Ottoni5Daniela Ferreira Domingos6Rodrigo Gouvea Taketani7Valéria Maia de Oliveira8André Oliveira de Souza Lima9João Lucio Azevedo10Jorge Luiz Mazza Rodrigues11Fernando Dini Andreote12Center of Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, São Paulo 13400-970, BrazilLuiz de Queiroz College of Agriculture, University of São Paulo, Piracicaba, São Paulo 13418-900, BrazilLuiz de Queiroz College of Agriculture, University of São Paulo, Piracicaba, São Paulo 13418-900, BrazilDepartment of Biology, University of Texas, Arlington, TX 76019, USAFleury Group, Jabaquara, São Paulo, 04344-070, BrazilCenter for Chemical, Biological and Agricultural Research (CPQBA), University of Campinas, Campinas, São Paulo 13148-218, BrazilCenter for Chemical, Biological and Agricultural Research (CPQBA), University of Campinas, Campinas, São Paulo 13148-218, BrazilEmbrapa Environment, Jaguariúna, São Paulo 13820-000, BrazilCenter for Chemical, Biological and Agricultural Research (CPQBA), University of Campinas, Campinas, São Paulo 13148-218, BrazilDepartment of Biological Sciences, University of Vale do Itajai, Itajai, Santa Catarina 88302-202, BrazilLuiz de Queiroz College of Agriculture, University of São Paulo, Piracicaba, São Paulo 13418-900, BrazilDepartment of Land, Air and Water Resources, University of California, Davis, CA 95616, USALuiz de Queiroz College of Agriculture, University of São Paulo, Piracicaba, São Paulo 13418-900, BrazilThe 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.https://www.mdpi.com/2306-5354/4/3/62bioprospectionenzyme characterizationhydrolasesmicrobial communities3D modeling |
| spellingShingle | Fábio Lino Soares Joelma Marcon Michele de Cássia Pereira e Silva Nittaya Khakhum Louise Teixeira Cerdeira Júlia Ronzella Ottoni Daniela Ferreira Domingos Rodrigo Gouvea Taketani Valéria Maia de Oliveira André Oliveira de Souza Lima João Lucio Azevedo Jorge Luiz Mazza Rodrigues Fernando Dini Andreote A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove Bioengineering bioprospection enzyme characterization hydrolases microbial communities 3D modeling |
| 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 | bioprospection enzyme characterization hydrolases microbial communities 3D modeling |
| url | https://www.mdpi.com/2306-5354/4/3/62 |
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