Journal
ELECTRONIC JOURNAL OF BIOTECHNOLOGY
Volume 24, Issue -, Pages 56-62Publisher
UNIV CATOLICA DE VALPARAISO
DOI: 10.1016/j.ejbt.2016.09.004
Keywords
Protein modeling; Site-directed mutagenesis; Cellulases; Recombinant endoglucanase gene; Catalytic domain of glycoside hydrolase; Carbohydrate-binding module; High pH stability; Tolerance to deleterious chemicals; Tolerance to heavy metals; Tolerance to detergents
Categories
Funding
- Key Laboratory of Biotic Environment and Ecology Safety in Anhui Province
- Anhui Provincial Natural Science Foundation [1208085QC56]
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Background: Endoglucanase, one of three type cellulases, can randomly cleave internal beta-1,4-linkages in cellulose polymers. Thus, it could be applied in agricultural and industrial processes. Results: A novel endoglucanase gene (JqCel5A) was cloned from Jonesia quinghaiensis and functionally expressed in Escherichia coli Rosetta (DE3). It contained 1722 bp and encoded a 573-residue polypeptide consisting of a catalytic domain of glycoside hydrolase family 5 (GH5) and a type 2 carbohydrate-binding module (CBM2), together with a predicted molecular mass of 61.79 kD. The purified JqCel5A displayed maximum activity at 55 degrees C and pH 7.0, with 21.7 U/mg, 26.19 U/mg and 4.81 U/mg towards the substrate carboxymethyl cellulose, barley glucan and filter paper, respectively. Interestingly, JqCel5A exhibited high pH stability over a broad pH range of pH (3-11), and had good tolerance to a wide variety of deleterious chemicals including heavy metals and detergent. The catalytic mechanism of JqCel5A was also investigated by site mutagenesis and homology-modeling in this study. Conclusions: It was believed that these properties might make JqCel5A to be potentially used in the suitable industrial catalytic condition, which has a broad pH fluctuation and/or chemical disturbance. (C) 2016 Pontificia Universidad Catalica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.
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