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Cloning and characterization of a novel GH44 family endoglucanase from mangrove soil metagenomic library

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Abstract

A novel endoglucanase gene, mgcel44, was isolated from a mangrove soil metagenomic library by functional-based screening. It encodes a 648-aa peptide with a catalytic domain of glycosyl hydrolase family 44. The deduced amino acid sequence of mgcel44 shares less than 50 % identity with endoglucanases in GenBank database. mgcel44 was cloned and overexpressed in Escherichia coli. The recombinant enzyme, MgCel44, has a molecular mass of 70.8 kDa as determined by SDS-PAGE. Its optimal pH and temperature for activity were 6 and 45 °C, respectively. It was highly active at 25–45 °C and pH 5–8. Its activity was enhanced in 0.5 M NaCl by >1.6-fold and stable up to 1.5 M NaCl. MgCel44 was resistant to several organic solvents and had high activity at 15 % (v/v) solvent after incubating for 24 h at 25 °C.

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Acknowledgments

This study was supported by the National Basic Research Program of China (973 Program, 2010CB833801), National Natural Science Foundation of China (Grant No. 41230962).

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Correspondence to Si Zhang.

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Mai, Z., Su, H., Yang, J. et al. Cloning and characterization of a novel GH44 family endoglucanase from mangrove soil metagenomic library. Biotechnol Lett 36, 1701–1709 (2014). https://doi.org/10.1007/s10529-014-1531-4

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  • DOI: https://doi.org/10.1007/s10529-014-1531-4

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