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Expression and Characterization of a Novel Antifungal Exo-β-1,3-glucanase from Chaetomium cupreum

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Abstract

A novel β-1,3-glucanase gene, designated Ccglu17A, was cloned from the biological control fungus Chaetomium cupreum Ame. Its 1626-bp open reading frame encoded 541 amino acids. The corresponding amino acid sequence showed highest identity (67 %) with a glycoside hydrolase family 17 β-1,3-glucanase from Chaetomium globosum. The recombinant protein Ccglu17A was successfully expressed in Pichia pastoris, and the enzyme was purified to homogeneity with 10.1-fold purification and 47.8 % recovery yield. The protein’s molecular mass was approximately 65 kDa, and its maximum activity appeared at pH 5.0 and temperature 45 °C. Heavy metal ions Fe2+, Mn2+, Cu2+, Co2+, Ag+, and Hg2+ had inhibitory effects on Ccglu17A, but Ba2+ promoted the enzyme’s activity. Ccglu17A exhibited high substrate specificity, almost exclusively catalyzing β-1,3-glycosidic bond cleavage in various polysaccharoses to liberate glucose. The enzyme had a Km of 2.84 mg/mL and Vmax of 10.7 μmol glucose/min/mg protein for laminarin degradation under optimal conditions. Ccglu17A was an exoglucanase with transglycosylation activity based on its hydrolytic properties. It showed potential antifungal activity with a degradative effect on cell walls and inhibitory action against the germination of pathogenic fungus. In conclusion, Ccglu17A is the first functional exo-1,3-β-glucanase to be identified from C. cupreum and has potential applicability in industry and agriculture.

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Acknowledgements

The authors wish to thank the Chinese government for the financial support of this study under the National High Technology Research and Development Program (2011AA10A205) and “Twelfth Five-Year Plan” National Science and Technology Program on Rural Area (2014BAL02B00).

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Authors and Affiliations

  1. School of Life Science and Technology, Harbin Institute of Technology, Nangang District, Harbin, People’s Republic of China

    Cheng Jiang, Jinzhu Song, Hua Cong & Qian Yang

  2. School of Chemical Engineering and Technology, Harbin Institute of Technology, Nangang District, Harbin, People’s Republic of China

    Junzheng Zhang

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  1. Cheng Jiang
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  2. Jinzhu Song
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  3. Hua Cong
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  4. Junzheng Zhang
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Correspondence to Junzheng Zhang.

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Jiang, C., Song, J., Cong, H. et al. Expression and Characterization of a Novel Antifungal Exo-β-1,3-glucanase from Chaetomium cupreum . Appl Biochem Biotechnol 182, 261–275 (2017). https://doi.org/10.1007/s12010-016-2325-z

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  • DOI: https://doi.org/10.1007/s12010-016-2325-z

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