Identification, cloning, and characterization of a novel chitinase from leaf-cutting ant Atta sexdens: An enzyme with antifungal and insecticidal activity

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Micocci, Kelli C. ; Moreira, Ariele C. ; Sanchez, Amanda D. ; Pettinatti, Jessica L. ; Rocha, Marina C. ; Dionizio, Bruna S. ; Correa, Katia C. S. ; Malavazi, Iran ; Wouters, Felipe C. ; Bueno, Odair C. ; Souza, Dulce Helena F.

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1867, n. 1, p. 10-pg., 2023-01-01.

Resumo

Chitinases are enzymes that degrade chitin, a polysaccharide found in the exoskeleton of insects, fungi, yeast, and internal structures of other vertebrates. Although chitinases isolated from bacteria, fungi and plants have been reported to have antifungal or insecticide activities, chitinases from insects with these activities have been sel-domly reported. In this study, a leaf-cutting ant Atta sexdens DNA fragment containing 1623 base pairs was amplified and cloned into a vector to express the protein (AsChtII-C4B1) in Pichia pastoris. AsChtII-C4B1, which contains one catalytic domain and one carbohydrate-binding module (CBM), was secreted to the extracellular medium and purified by ammonium sulfate precipitation followed by nickel column chromatography. AsChtII-C4B1 showed maximum activity at pH 5.0 and 55 degrees C when tested against colloidal chitin substrate and main-tained >60% of its maximal activity in different temperatures during 48 h. AsChtII-C4B1 decreased the survival of Spodoptera frugiperda larvae fed with an artificial diet that contained AsChtII-C4B1. Our results have indicated that AsChtII-C4B1 has a higher effect on larva-pupa than larva-larva molts. AsChtII-C4B1 activity targets more specifically the growth of filamentous fungus than yeast. This work describes, for the first time, the obtaining a recombinant chitinase from ants and the characterization of its insecticidal and antifungal activities. (AU)

• Processo FAPESP: 12/25299-6 - Estudos integrados para o controle de formigas cortadeiras
• Beneficiário: João Batista Fernandes
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/06198-8 - Estudo funcional de acetilcolinesterase (AChE) e outras enzimas de Atta Sexdens rubropilosa e seu fungo simbionte Leucoagaricus gongylophorus, imobilizações das mesmas em eletrodo e busca de inibidores específicos
• Beneficiário: Kelli Cristina Micocci
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/19694-3 - Identificação do papel das proteínas de choque térmico HsfA e Hsp90 e do regulador SmiA na atividade da via da integridade da parede celular no fungo patogênico oportunista humano Aspergillus fumigatus
• Beneficiário: Iran Malavazi
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 18/06297-9 - Produção e caracterização de proteínas quiméricas contendo domínio catalítico quitinase e módulos para ligação em quitina
• Beneficiário: Dulce Helena Ferreira de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular