RT Journal Article T1 Insight into novel biochemical features and function of novel antifungal GH16-β-glucanase from Bacillus velezensis A1 Vela-Corcia, David A1 De-Vicente-Moreno, Antonio A1 Pérez-García, Alejandro A1 Romero-Hinojosa, Diego Francisco K1 Celulasa K1 Plantas - Enfermedades criptogámicas AB β-Glucanases play a critical role in degrading fungal cell wall β-glucans, compromising fungal integrity and development. Here we characterize BvlzGluc, a novel β-glucanase from Bacillus velezensis CECT 8237 with potent and specific activity against β-1,3-glucans. The enzyme hydrolyzes yeast-derived β-glucan but is inactive against β-1,6- and β-1,3/1,4-linked polysaccharides, such as laminarin and lichenin, indicating strict substrate specificity. Mass spectrometry of Botrytis cinerea cell wall extracts revealed that BvlzGluc cleaves high-molecular-weight β-glucans into smaller oligosaccharides, supporting its classification as an endo-β-1,3-glucanase. Kinetic analysis yielded a Km of 48 μM and a Vmax of 0.026 μmol/min. Structural modeling showed a GH16 jelly roll fold lacking a classical catalytic groove, and site-directed mutagenesis of two predicted binding residues (Y36 and T216) confirmed their functional relevance. CD spectroscopy revealed that T216A disrupts folding, while Y36A retains structure but shows reduced activity. Despite its compact architecture and divergence from canonical motifs, BvlzGluc demonstrates efficient antifungal function. Its physicochemical simplicity and specificity make it a promising candidate for scalable production and targeted applications in crop protection. This work expands our understanding of bacterial glucanases and underscores the value of structurally minimal enzymes in biocontrol strategies. PB Elsevier YR 2026 FD 2026 LK https://hdl.handle.net/10630/45130 UL https://hdl.handle.net/10630/45130 LA eng NO D. Vela-Corcia, A. de Vicente, A. Pérez-García, D. Romero, Insight into novel biochemical features and function of novel antifungal GH16-β-glucanase from Bacillus velezensis, International Journal of Biological Macromolecules, Volume 340, Part 1, 2026, 150106, ISSN 0141-8130, https://doi.org/10.1016/j.ijbiomac.2026.150106. NO Funding for open access charge: Universidad de Málaga / CBUA DS RIUMA. Repositorio Institucional de la Universidad de Málaga RD 25 feb 2026