Biochemical characterization of two novel β-glucosidase genes by metagenome expression cloning
| Autor: | Qin Wang, Xian-Lai Tang, Bo Wu, Shuang-Xi Li, Feng-Feng Luo, Ke Jin, Lan-Lan Wu, Ge-Fei Ma, Zhen-Yu Hao, Chengjian Jiang, Gao-Chao Zhao, Peihong Shen |
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| Rok vydání: | 2011 |
| Předmět: |
Environmental Engineering
Molecular Sequence Data Gene Expression Bioengineering Biology law.invention Hydrolysis chemistry.chemical_compound Glucoside law Amino Acid Sequence Enzyme kinetics Cloning Molecular Waste Management and Disposal Soil Microbiology chemistry.chemical_classification Base Sequence Renewable Energy Sustainability and the Environment beta-Glucosidase Substrate (chemistry) General Medicine Enzyme chemistry Biochemistry DNA glycosylase Expression cloning Recombinant DNA Metagenome |
| Zdroj: | Bioresource Technology. 102:3272-3278 |
| ISSN: | 0960-8524 |
| DOI: | 10.1016/j.biortech.2010.09.114 |
| Popis: | Two novel β-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 β-glucosidases. Functional characterization through high-performance liquid chromatography demonstrated that purified recombinant Bgl1D and Bgl1E proteins hydrolyzed d -glucosyl-β-(1–4)- d -glucose to glucose. Using p-nitrophenyl-β- d -glucoside as substrate, Km was 0.54 and 2.11 mM, and kcat/Km 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 AlCl3 and LiCl. Bgl1D also displayed remarkable activity across a broad pH range (5.5–10.5), making it a potential candidate for industrial applications. |
| Databáze: | OpenAIRE |
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