Chitinases catalyze the hydrolysis of β-1,4-glycosidic bonds in chitin, a structural biopolymer synthesized by numerous organisms. Although these enzymes have been widely investigated, studies focusing on insect-derived chitinases remain limited. In this study, three recombinant chitinases from the leaf-cutter ant
Atta sexdens were
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Chitinases catalyze the hydrolysis of β-1,4-glycosidic bonds in chitin, a structural biopolymer synthesized by numerous organisms. Although these enzymes have been widely investigated, studies focusing on insect-derived chitinases remain limited. In this study, three recombinant chitinases from the leaf-cutter ant
Atta sexdens were cloned, expressed in
Pichia pastoris, and biochemically characterized. The enzymes-AsChtII-C2B3 (one catalytic and three chitin-binding domains), AsChtII-C3C4 (two catalytic domains), and AsChtII-C5B1 (one catalytic and one binding domain), exhibited optimal activity at pH 4–5 and 50 °C using colloidal chitin as substrate. Chitinase activity on colloidal α-chitin was confirmed by
1H NMR (proton nuclear magnetic resonance) spectroscopy, revealing GlcNAc concentrations of 0.41, 0.48, and 0.56 mmol L
−1 for AsChtII-C3C4, AsChtII-C2B3, and AsChtII-C5B1, respectively. Their antifungal activities were evaluated against the human pathogens
Candida albicans and
Aspergillus fumigatus, as well as the phytopathogen
Lasiodiplodia theobromae. Distinct inhibition profiles were observed: AsChtII-C5B1 (150 µg/mL) showed the highest activity against
C. albicans (87.6% inhibition), while AsChtII-C3C4 (25 µg/mL) was most effective against
A. fumigatus (60% inhibition). Notably, only AsChtII-C2B3 inhibited
L. theobromae growth, inducing severe hyphal deformations observed by scanning electron microscopy (SEM). These findings demonstrate that recombinant
A. sexdens chitinases exhibit species-specific antifungal properties, underscoring their potential as biotechnological tools for medical and agricultural applications.
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