Biochemical characterization of two novel ß-glucosidase genes by metagenome expression cloning.
Bioresour Technol
; 102(3): 3272-8, 2011 Feb.
Article
em En
| MEDLINE
| ID: mdl-20971635
ABSTRACT
Two novel ß-glucosidase genes designated as bgl1D and bgl1E, which encode 172- and 151-aa peptides, respectively, were cloned by function-based screening of a metagenomic library from uncultured soil microorganisms. Sequence analyses indicated that Bgl1D and Bgl1E exhibited lower similarities with some putative ß-glucosidases. Functional characterization through high-performance liquid chromatography demonstrated that purified recombinant Bgl1D and Bgl1E proteins hydrolyzed D-glucosyl-ß-(1-4)-D-glucose to glucose. Using p-nitrophenyl-ß-D-glucoside as substrate, K(m) was 0.54 and 2.11 mM, and k(cat)/K(m) was 1489 and 787 mM(-1) min(-1) for Bgl1D and Bgl1E, respectively. The optimum pH and temperature for Bgl1D was pH 10.0 and 30°C, while the optimum values for Bgl1E were pH 10.0 and 25°C. Bgl1D exhibited habitat-specific characteristics, including higher activity in lower temperature and at high concentrations of AlCl(3) and LiCl. Bgl1D also displayed remarkable activity across a broad pH range (5.5-10.5), making it a potential candidate for industrial applications.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Microbiologia do Solo
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Beta-Glucosidase
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Metagenoma
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article