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Cloning, expression, and characterization of a thermostable GH7 endoglucanase from Myceliophthora thermophila capable of high-consistency enzymatic liquefaction

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Abstract

An endoglucanase gene from the thermophilic fungus Myceliophthora thermophila, belonging to the glycoside hydrolase family 7, was functionally expressed in methylotrophic yeast Pichia pastoris. The putative endoglucanase from the genomic DNA was successfully cloned in P. pastoris X-33 and the recombinant enzyme was purified to its homogeneity (65 kDa) and subsequently characterized. Substrate specificity analysis revealed that the enzyme exhibits high activity on substrates containing β-1,4-glycosidic bonds such as carboxymethyl cellulose, barley β-glucan, and cello-oligosaccharides, as well as activity on xylan-containing substrates, including arabinoxylan and oat spelt xylan. MtEG7a was proved to liquefy rapidly and efficiently pretreated wheat straw, indicating its key role to the initial step of hydrolysis of high-solids lignocellulose substrates. High thermostability of the endoglucanase reflects potential commercial significance of the enzyme.

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Acknowledgments

Anthi Karnaouri’s work was supported by grant from the Hellenic State Scholarships Foundation. The research leading to these results has received funding from the European Community’s 7th Framework Programme (FP7/2007-2013), under grant agreement 213139—the HYPE project. The authors would like to thank Thomas Paschos for his help in liquefaction experiments.

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Authors and Affiliations

  1. Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, 5 Iroon Polytechniou Str, Zografou Campus, 15700, Athens, Greece

    Anthi C. Karnaouri & Evangelos Topakas

  2. Biochemical and Chemical Process Engineering, Division of Sustainable Process Engineering, Department of Civil, Environmental and Natural Resources Engineering, Luleå University of Technology, SE-97187, Luleå, Sweden

    Paul Christakopoulos

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  1. Anthi C. Karnaouri
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  2. Evangelos Topakas
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Correspondence to Paul Christakopoulos.

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Anthi C. Karnaouri and Evangelos Topakas equally contributed to this article.

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Karnaouri, A.C., Topakas, E. & Christakopoulos, P. Cloning, expression, and characterization of a thermostable GH7 endoglucanase from Myceliophthora thermophila capable of high-consistency enzymatic liquefaction. Appl Microbiol Biotechnol 98, 231–242 (2014). https://doi.org/10.1007/s00253-013-4895-9

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