TY - JOUR
T1 - A novel multifunctional β-N-acetylhexosaminidase revealed through metagenomics of an oil-spilled mangrove
AU - Soares, Fábio Lino
AU - Marcon, Joelma
AU - E Silva, Michele de Cássia Pereira
AU - Khakhum, Nittaya
AU - Cerdeira, Louise Teixeira
AU - Ottoni, Júlia Ronzella
AU - Domingos, Daniela Ferreira
AU - Taketani, Rodrigo Gouvea
AU - de Oliveira, Valéria Maia
AU - Lima, André Oliveira de Souza
AU - Azevedo, João Lucio
AU - Rodrigues, Jorge Luiz Mazza
AU - Andreote, Fernando Dini
PY - 2017/9
Y1 - 2017/9
N2 - 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.
AB - 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.
KW - 3D modeling
KW - Bioprospection
KW - Enzyme characterization
KW - Hydrolases
KW - Microbial communities
UR - http://www.scopus.com/inward/record.url?scp=85053135402&partnerID=8YFLogxK
U2 - 10.3390/bioengineering4030062
DO - 10.3390/bioengineering4030062
M3 - Article
AN - SCOPUS:85053135402
SN - 2306-5354
VL - 4
JO - Bioengineering
JF - Bioengineering
IS - 3
M1 - 62
ER -