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Trans-glycosylation capacity of a highly glycosylated multi-specific β-glucosidase from Fusarium solani

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Abstract

An extracellular β-glucosidase from Fusaruim solani cultivated on wheat bran was purified by only two chromatographic steps. The purified enzyme exhibited optimal temperature and pH at 60 °C and pH 5, respectively. The purified β-glucosidase behaves as a very large protein due to its high degree of glycosylation. More interestingly, the endoglycosidase H (Endo H) treatment led to 97.55% loss of its initial activity after 24 h of treatment. Besides, the addition of Tunicamycin (nucleoside antibiotic blocking the N-glycosylation first step) during the culture of the fungus affected seriously the glycosylation of the enzyme. Both treatments (endo H and Tunicamycin) strengthened the idea that the hyperglycosylation is involved in the β-glucosidase activity and thermostability. This enzyme was also shown to belong to class III of β-glucosidases (multi-specific) since it was able to act on either cellobiose, gentiobiose or sophorose which are disaccharide composed of two units of d-glucose connected by β1–4, β1–6 and β1–2 linkage, respectively. The β-glucosidase activity was strongly enhanced by ferrous ion (Fe2+) and high ionic strength (1 M KCl). The purified enzyme exhibited an efficient transglycosylation capacity allowing the synthesis of cellotriose and cellotetraose using cellobiose as donor.

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Acknowledgements

This work was supported by grants from the Ministry of Higher Education, Scientific Research and Technology, Tunisia. The authors would like to thank Ines Belhadj for helpful discussions and advices, Hajer Hassairi and Kamel Walha for their expert help in chromatographic analysis and finally Abdelmajid Dammak for his proof reading of the text.

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Authors and Affiliations

  1. Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia

    Manel Boudabbous, Ines Ben Hmad, Mariem Mssawra, Hafedh Belghith & Ali Gargouri

  2. Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia

    Walid Saibi

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  1. Manel Boudabbous
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  2. Ines Ben Hmad
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Correspondence to Manel Boudabbous.

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Boudabbous, M., Ben Hmad, I., Saibi, W. et al. Trans-glycosylation capacity of a highly glycosylated multi-specific β-glucosidase from Fusarium solani . Bioprocess Biosyst Eng 40, 559–571 (2017). https://doi.org/10.1007/s00449-016-1721-7

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