Biochemical characterization of a Kunitz inhibitor from Inga edulis seeds with antifungal activity against Candida spp.

Biochemical characterization of a Kunitz inhibitor from Inga edulis seeds with antifungal activity against Candida spp.

We describe the characterization of IETI, the first trypsin inhibitor purified from Inga edulis, a tree widely distributed in Brazil. Two-step chromatography was used to purify IETI, a protein composed of a single peptide chain of 19,685.10 Da. Amino-terminal sequencing revealed that IETI shows homology with the Kunitz family, as substantiated by its physical?chemical features, such as its thermal (up to 70 °C) and wide-range pH stability (from 2 to 10), and the value of its dissociation constant (6.2 nM). IETI contains a single reactive site for trypsin, maintained by a disulfide bridge; in the presence of DTT, its inhibitory activity was reduced in a time- and concentration-dependent manner. IETI presented activity against Candida ssp., including C. buinensis and C. tropicalis. IETI inhibitory activity triggered yeast membrane permeability, affecting cell viability, thus providing support for the use of IETI in further studies for the control of fungal infections.

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Bibliographic Details
Main Authors: DIB, H. X., OLIVEIRA, D. G. L. de, OLIVEIRA, C. F. R. de, TAVEIRA, G. B., MELLO, E. de O., VERBISCK, N. V., CHANG, M. R., CORRÊA JUNIOR, D., GOMES, V. M., MACEDO, M. L. R.
Other Authors: Heloisa Xavier Dib, Universidade Federal do Mato Grosso do Sul - UFMS/Faculdade de Ciências de Alimentos e Nutrição/Laboratório de Purificação de Proteínas e suas Funções Biológicas; Daniella Gorete Lourenço de Oliveira, Universidade Federal do Mato Grosso do Sul - UFMS/Faculdade de Ciências de Alimentos e Nutrição/Laboratório de Purificação de Proteínas e suas Funções Biológicas; Caio Fernando Ramalho de Oliveira, Universidade Federal do Mato Grosso do Sul - UFMS/Faculdade de Ciências de Alimentos e Nutrição/Laboratório de Purificação de Proteínas e suas Funções Biológicas; Gabriel Bonan Taveira, Universidade Estadual do Norte Fluminense - UENF/Laboratório de Fisiologia e Bioquímica de Microrganismos; Erica de Oliveira Mello, Universidade Estadual do Norte Fluminense - UENF/Laboratório de Fisiologia e Bioquímica de Microrganismos; NEWTON VALERIO VERBISCK, CNPGC; Marilene Rodrigues Chang, Universidade Federal do Mato Grosso do Sul - UFMS/Faculdade de Ciências de Alimentos e Nutrição/Laboratório de Pesquisas Microbiológicas; Dario Corrêa Junior; Valdirene Moreira Gomes, Universidade Estadual do Norte Fluminense - UENF/Laboratório de Fisiologia e Bioquímica de Microrganismos; Maria Lígia Rodrigues Macedo, Universidade Federal do Mato Grosso do Sul - UFMS/Faculdade de Ciências de Alimentos e Nutrição/Laboratório de Purificação de Proteínas e suas Funções Biológicas.
Format: Artigo de periódico biblioteca
Language:pt_BR
por
Published: 2019-02-18
Subjects:Candida buinensis, Trypsin inhibitors, Membrane permeability, Candida tropicalis,
Online Access:http://www.alice.cnptia.embrapa.br/alice/handle/doc/1106185
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Summary:We describe the characterization of IETI, the first trypsin inhibitor purified from Inga edulis, a tree widely distributed in Brazil. Two-step chromatography was used to purify IETI, a protein composed of a single peptide chain of 19,685.10 Da. Amino-terminal sequencing revealed that IETI shows homology with the Kunitz family, as substantiated by its physical?chemical features, such as its thermal (up to 70 °C) and wide-range pH stability (from 2 to 10), and the value of its dissociation constant (6.2 nM). IETI contains a single reactive site for trypsin, maintained by a disulfide bridge; in the presence of DTT, its inhibitory activity was reduced in a time- and concentration-dependent manner. IETI presented activity against Candida ssp., including C. buinensis and C. tropicalis. IETI inhibitory activity triggered yeast membrane permeability, affecting cell viability, thus providing support for the use of IETI in further studies for the control of fungal infections.

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