Isolation and Characterization of a Glycosyl Hydrolase Family 16 ß-Agarase from a Mangrove Soil Metagenomic Library.
Int J Mol Sci
; 17(8)2016 Aug 19.
Article
em En
| MEDLINE
| ID: mdl-27548158
ABSTRACT
A mangrove soil metagenomic library was constructed and a ß-agarase gene designated as AgaML was isolated by functional screening. The gene encoded for a 659-amino-acids polypeptide with an estimated molecular mass of 71.6 kDa. The deduced polypeptide sequences of AgaML showed the highest identity of 73% with the glycoside hydrolase family 16 ß-agarase from Microbulbifer agarilyticus in the GenBank database. AgaML was cloned and highly expressed in Escherichia coli BL21(DE3). The purified recombinant protein, AgaML, showed optimal activity at 50 °C and pH 7.0. The kinetic parameters of Km and Vmax values toward agarose were 4.6 mg·mL(-1) and 967.5 µM·min(-1)·mg(-1), respectively. AgaML hydrolyzed the ß-1,4-glycosidic linkages of agar to generate neoagarotetraose (NA4) and neoagarohexaose (NA6) as the main products. These characteristics suggest that AgaML has potential application in cosmetic, pharmaceuticals and food industries.
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1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Glicosídeo Hidrolases
Idioma:
En
Revista:
Int J Mol Sci
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
China