Cloning and characterization of a new β-Glucosidase from a metagenomic library of Rumen of cattle feeding with Miscanthus sinensis

BMC Biotechnology

Yadan Li¹  Ning Liu³  Hui Yang¹  Fei Zhao¹  Ye Yu²  Yun Tian¹  Xiangyang Lu¹ 

[1] Hunan Agricultural Bioengineering Research Institute, Changsha 410128, China

[2] Departments of Physiology, Biochemistry, and Molecular Cell Biology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

[3] College of Life Science, Hunan Normal University, Changsha 410181, China

关键词: Metagenomic library;    Miscanthus sinensis;    Rumen;    β-glucosidase;   

Others  :  1084544
DOI  :  10.1186/1472-6750-14-85

 received in 2014-07-09, accepted in 2014-09-24,  发布年份 2014

【 摘 要 】

Background

The study on the second generation bio-fuel is a hot area of current research of renewable energy. Among series of key points in this area, the role of β-glucosidase in the degradation of intermediate gluco-oligosaccharides limits the rate of the complete saccharification of lignocellulose.

Results

In this study, a new β-glucosidase gene, unglu135B12, which was isolated from a metagenomic library of rumen of cattle feeding with Miscanthus sinensis by the function-based screening, encodes a 779 amino acid polypeptide that contains a catalytic domain belonging to glycoside hydrolase family 3 (GH3). It was recombinantly expressed, purified and biochemically characterized. The recombinant β-glucosidase, unglu135B12, displayed optimum enzymatic activity at pH 5.0 at 38°C, and showed the highest specific activity of 2.5 × 10³ U/mg under this optimal condition to p-nitrophenyl-β-D-glucopyranoside (pNPG), and its Km and Vmax values were 0.309 mmol/L and 7.292 μmol/min, respectively. In addition, the presence of Ca²⁺, K⁺, Na⁺ slightly improved β-glucosidase activity of unglu135B12 by about 5%, while about 10 ~ 85% loss of β-glucosidase activity was induced by addition of Mn²⁺, Fe³⁺, Zn²⁺, Cu²⁺. Interestingly, unglu135B12 was activated by glucose at the concentration lower than 40 mM.

Conclusions

Our findings indicate that unglu135B12 is a new β-glucosidase derived from rumen of cattle, and it might be a potent candidate for saccharification of lignocellulose in industrial application.

【 授权许可】

   
2014 Li et al.; licensee BioMed Central Ltd.

【 图 表 】

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