Abstract

PARTIAL CHARACTERIZATION OF A IMMUNE-RELATED LIPID TRANSFER PROTEIN (LTP) AND ACTIVITY AGAINST YEASTS OF MEDICAL INTEREST FROM PROTEINS OF CANAVALIA ENSIFORMES (JACK-BEAN) SEEDS

Marciele Souza da Silva, Suzanna de Fátima Ferreira Ribeiro,  Gabriel Bonan Taveira, Érica de Oliveira Mello, André de Oliveira Carvalho, Eduardo Alves Gamosa de Oliveira, Antonia Elenir Amâncio Oliveira, Viviane Veiga do Nascimento, Valdirene Moreira Gomes*

Abstract

The aim of this work was to characterize antimicrobial peptides fromCanavalia ensiformis seeds and evaluate their antimicrobial activityagainst fungi. The crude extract was subjected to anion-exchangechromatography in a DEAE-Sepharose column, resulting in twofractions called D1 and D2. The D1 basic fraction was subjected toreverse-phase chromatography on a C2/C18 column, where it wasseparated into six new fractions named (RP1, RP2, RP3, RP4, RP5 andRP6). The entire purification process was monitored using tricine gelelectrophoresis. In D1 fraction a lipid transfer protein (LTP) immunerelated protein were also detected by ELISA. The D1 fraction inhibitedthe growth of the yeasts Candida albicans, Candida tropicalis andPichia membranifaciens and promoted changes in cellularagglomeration in some of the yeasts tested, including the formation ofpseudohyphae in C. albicans. In the "spot assay", different degrees ofsensitivity were observed, indicating that the yeast C. albicans has afungicidal effect and that the other tested strains have a fungistatic effect. Membranepermeabilization was assessed, and when observed with a fluorescence microscope, all yeast species showed fluorescence in the presence of the D1 fraction. It was also shown that this fraction inhibits the acidification of the medium of Saccharomyces cerevisiae and C. albicans.

Keywords: Antimicrobial peptides, yeast, LTP, fungi.