A novel multifunctional β-N-acetylhexosaminidase revealed through metagenomics of an oil-spilled mangrove

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

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7 Scopus citations

Abstract

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.

Original languageEnglish (US)
Article number62
JournalBioengineering
Volume4
Issue number3
DOIs
StatePublished - Sep 2017
Externally publishedYes

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Keywords

  • 3D modeling
  • Bioprospection
  • Enzyme characterization
  • Hydrolases
  • Microbial communities

ASJC Scopus subject areas

  • Bioengineering

Cite this

Soares, F. L., Marcon, J., E Silva, M. D. C. P., Khakhum, N., Cerdeira, L. T., Ottoni, J. R., Domingos, D. F., Taketani, R. G., de Oliveira, V. M., Lima, A. O. D. S., Azevedo, J. L., Rodrigues, J. L. M., & Andreote, F. D. (2017). A novel multifunctional β-N-acetylhexosaminidase revealed through metagenomics of an oil-spilled mangrove. Bioengineering, 4(3), [62]. https://doi.org/10.3390/bioengineering4030062