Afil, a Lectin from Aplysina fistularis, Exhibits Antibiofilm and Synergistic Antibacterial Activity Against Resistant Bacteria.

Lectins from marine sponges have emerged as promising candidates for antimicrobial strategies, particularly against biofilm-forming pathogens. In this study, we report the purification, biochemical characterization, and antibiofilm properties of a new lectin (AfiL) isolated from Aplysina fistularis. AfiL exhibited typical features of sponge lectins, including a β-sheet-rich secondary structure and a predominant oligomeric state in solution. Dynamic light scattering (DLS) analyses confirmed that AfiL predominantly exists as a well-defined oligomer at acidic and neutral pH. Sequence analysis revealed similarity to a putative collectin-like protein from sponge Desydea avara. AfiL selectively agglutinated Staphylococcus aureus strains, correlating with its preferential binding to lipoteichoic acid (LTA). The lectin demonstrated significant antibiofilm activity against S. aureus, S. epidermidis, and Escherichia coli strains, and exhibited synergistic or additive effects when combined with conventional antibiotics against a Methicillin-resistant S. aureus. Isothermal titration calorimetry (ITC) revealed a strong interaction between AfiL and porcine stomach mucin (Kd = 1.71 × 10-6 M), consistent with multivalent carbohydrate recognition. Overall, our findings highlight the potential of AfiL as a novel antibiofilm agent with species-specific modulatory effects on antibiotic activity and provide new insights into the functional versatility of sponge-derived lectins in microbial control strategies.