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A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove.

Fábio Lino SoaresJoelma MarconMichele de Cássia Pereira E SilvaNittaya KhakhumLouise Teixeira CerdeiraJúlia Ronzella OttoniDaniela Ferreira DomingosRodrigo Gouvea TaketaniValéria Maia de OliveiraAndré Oliveira de Souza LimaJoão Lúcio AzevedoJorge Luiz Mazza RodriguesFernando Dini Andreote
Published in: Bioengineering (Basel, Switzerland) (2017)
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.
Keyphrases
  • genome wide
  • amino acid
  • copy number
  • heavy metals
  • hydrogen peroxide
  • gene expression
  • drug delivery
  • drinking water
  • case report
  • human health
  • single molecule
  • climate change
  • silver nanoparticles