title: Trans-glycosylation capacity of a highly glycosylated multi-specific β-glucosidase from Fusarium solani.
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.
authors.affiliation: Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia. manelboudabous@gmail.com.
authors.forename: Manel
authors.initials: M
authors.lastname: Boudabbous
authors.affiliation: Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia.
authors.forename: Ines
authors.initials: I
authors.lastname: Ben Hmad
authors.affiliation: Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia.
authors.forename: Walid
authors.initials: W
authors.lastname: Saibi
authors.affiliation: Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia.
authors.forename: Mariem
authors.initials: M
authors.lastname: Mssawra
authors.affiliation: Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia.
authors.forename: Hafedh
authors.initials: H
authors.lastname: Belghith
authors.affiliation: Laboratory of Molecular Biology of Eucaryotes, Centre of Biotechnology of Sfax (CBS), University of Sfax, BP 1177, 3018, Sfax, Tunisia.
authors.forename: Ali
authors.initials: A
authors.lastname: Gargouri
mesh_terms: D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D005670:Fusarium / Q000201:enzymology*; D006031:Glycosylation; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D005656:Fungal Proteins; D001617:beta-Glucosidase
keywords: Hyperglycosylation; Transglycosylation; β-Glucosidase