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A Novel Three Domains Glycoside Hydrolase Family 3 from Sclerotinia sclerotiorum Exhibits β-Glucosidase and Exoglucanase Activities: Molecular, Biochemical, and Transglycosylation Potential Analysis

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Abstract

The filamentous fungus Sclerotinia sclerotiorum produces a complete set of cellulolytic enzymes. We report here the purification and the biochemical characterization of a new β-glucosidase from S. sclerotiorum which belongs to the family 3 of glycoside hydrolases and that was named as SsBgl3. After two size-exclusion chromatography steps, purified protein bands of 80 and 90 kDa from SDS-PAGE were subjected to a mass spectrometry analysis. The results displayed four peptides from the upper band belonging to a polypeptide of 777 amino acids having a calculated molecular weight of 83.7 kDa. Biochemical analysis has been carried out to determine some properties. We showed that this SsBgl3 protein displayed both β-glucosidase and exoglucanase activities with optimal activity at 55 °C and at pH 5. The transglycosylation activity was investigated using gluco-oligosaccharides TLC analysis. The molecular modeling and comparison with different crystal structures of β-glucosidases showed that SsBgl3 putative protein present three domains. They correspond to an (α/β)8 domain TIM barrel, a five-stranded α/β sandwich domain (both of which are important for active-site organization), and a C-terminal fibronectin type III domain. Enzyme engineering will be soon investigated to identify the key residues for the catalytic reactions.

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Acknowledgments

The Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB) in the National Institute of Applied Sciences and Technology of Tunis, University of Carthage financed this work. The Tunisian Ministry of High Education, Scientific Research and Technology is gratefully acknowledged for the financial support of the training program. The laboratory of polymers, biopolymers and surfaces (PBS-UMR 6270) of Rouen is also gratefully acknowledged.

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Authors and Affiliations

  1. Laboratoire d’Ingénierie des Protéines et des Molécules Bioactives (LIP-MB), LR11ES24, National Institute of Applied Sciences and Technology, University of Carthage, 1080, Tunis Cedex, Tunisia

    Haifa Chahed, Aymen Ezzine & M. Nejib Marzouki

  2. Laboratoire Polymères Biopolymères Surfaces (PBS), UMR 6270 CNRS, Université de Rouen, 76821, Mont-Saint-Aignan Cedex, France

    Mohamed Amine Ben Mlouka, Christophe Rihouey, Julie Hardouin & Thierry Jouenne

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  1. Haifa Chahed
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  2. Aymen Ezzine
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Correspondence to Haifa Chahed.

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Chahed, H., Ezzine, A., Mlouka, M.A.B. et al. A Novel Three Domains Glycoside Hydrolase Family 3 from Sclerotinia sclerotiorum Exhibits β-Glucosidase and Exoglucanase Activities: Molecular, Biochemical, and Transglycosylation Potential Analysis. Mol Biotechnol 57, 993–1002 (2015). https://doi.org/10.1007/s12033-015-9892-z

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