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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 from
Canavalia ensiformis seeds and evaluate their antimicrobial activity
against fungi. The crude extract was subjected to anion-exchange
chromatography in a DEAE-Sepharose column, resulting in two
fractions called D1 and D2. The D1 basic fraction was subjected to
reverse-phase chromatography on a C2/C18 column, where it was
separated into six new fractions named (RP1, RP2, RP3, RP4, RP5 and
RP6). The entire purification process was monitored using tricine gel
electrophoresis. In D1 fraction a lipid transfer protein (LTP) immune
related protein were also detected by ELISA. The D1 fraction inhibited
the growth of the yeasts Candida albicans, Candida tropicalis and
Pichia membranifaciens and promoted changes in cellular
agglomeration in some of the yeasts tested, including the formation of
pseudohyphae in C. albicans. In the "spot assay", different degrees of
sensitivity were observed, indicating that the yeast C. albicans has a
fungicidal effect and that the other tested strains have a fungistatic effect. Membrane
permeabilization 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.


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