Characterization of a novel intracellular beta-1,3-Glucanase from Sclerotinia sclerotiorum and investigation of its biopesticide potential for application in sustainable agriculture

Mohamed, Safa Ben Hadj and Ezzine, Aymen and Tóth, Liliána and Galgóczi, László Norbert and Marzouki, Mohamed Najib and Smaali, Issam (2025) Characterization of a novel intracellular beta-1,3-Glucanase from Sclerotinia sclerotiorum and investigation of its biopesticide potential for application in sustainable agriculture. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 319 (4). No. 145795. ISSN 0141-8130

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Abstract

beta-1,3-glucanases are pivotal enzymes that have the capacity to degrade glucans in fungal cell walls. The present study focused on the characterization of a new intracellular beta-1,3-glucanase (SscleGH64) from the filamentous fungus Sclerotinia sclerotiorum. In silico and phylogenetic analysis permitted the classification of SscleGH64 within family 64 of glycoside hydrolases (GH64) and the GluB subfamily. Recombinant SscleGH64 was expressed in Escherichia coli BL21 and purified using a single-step process involving amberlite CG50 resin. The optimal conditions for SscleGH64 activity were determined to be pH 8 and 60 °C. The Km, Vmax and Kcat values of the purified enzyme for laminarin were 0.75 mg mL-1, 19.77 U mg-1 and 304 s-1, respectively. SscleGH64 hydrolyzed yeast and oats glucans, indicating its endo-type beta-1,3-glucanase activity, a property that is consistent with its predicted structural characteristics. SscleGH64 exhibited antifungal potential by the capacity to impede the germination of conidia, the proliferation of mycelia, and the formation of appressoria in phytopathogenic fungi, including Aspergillus niger, Botrytis cinerea, Cladosporium herbarum, and Fusarium oxysporum. Notably, SscleGH64 mitigated successfully the symptoms associated with B. cinerea infection in tomato leaves. Our findings suggest that SscleGH64 is a promising candidate for the inhibition of fungal diseases and the preservation of postharvest fruits. Copyright © 2025 Elsevier B.V. All rights reserved.

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