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Purification of an Antifungal Peptide from Seeds of Brassica oleracea var. gongylodes and Investigation of Its Antifungal Activity and Mechanism of Action.


ABSTRACT: In this study, a 8.5-kDa antifungal peptide designated as BGAP was purified from the crude extract of the seeds of Brassica oleracea var. gongylodes by employing a protocol that comprised cation exchange chromatography on SP-Sepharose, cation exchange chromatography on Mono S and gel filtration chromatography on Superdex peptide. BGAP showed the highest amino acid sequence similarity to defensin peptides by mass spectrometric analysis. BGAP showed a broad spectrum of antifungal activity with a half maximal inhibitory concentration at 17.33 ?g/mL, 12.37 ?g/mL, 16.81 ?g/mL, and 5.60 ?g/mL toward Colletotrichum higginsianum, Exserohilum turcicum, Magnaporthe oryzae and Mycosphaerella arachidicola, respectively. The antifungal activity of BGAP remained stable (i) after heat treatment at 40-100 °C for 15 min; (ii) after exposure to solutions of pH 1-3 and 11-13 for 15 min; (iii) after incubation with solutions containing K?, Ca2+, Mg2+, Mn2+ or Fe3+ ions at the concentrations of 20-150 mmol/L for 2 h; and (iv) following treatment with 10% methyl alcohol, 10% ethanol, 10% isopropanol or 10% chloroform for 2 h. Fluorescence staining experiments showed that BGAP brought about an increase in cell membrane permeability, a rise in reactive oxygen species production, a decrease in mitochondrial membrane potential, and an accumulation of chitin at the hyphal tips of Mycosphaerella arachidicola.

SUBMITTER: Wang C 

PROVIDER: S-EPMC6480268 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Purification of an Antifungal Peptide from Seeds of <i>Brassica oleracea</i> var. <i>gongylodes</i> and Investigation of Its Antifungal Activity and Mechanism of Action.

Wang Caicheng C   Zhang Yao Y   Zhang Weiwei W   Yuan Susu S   Ng Tzibun T   Ye Xiujuan X  

Molecules (Basel, Switzerland) 20190404 7


In this study, a 8.5-kDa antifungal peptide designated as BGAP was purified from the crude extract of the seeds of <i>Brassica oleracea</i> var. <i>gongylodes</i> by employing a protocol that comprised cation exchange chromatography on SP-Sepharose, cation exchange chromatography on Mono S and gel filtration chromatography on Superdex peptide. BGAP showed the highest amino acid sequence similarity to defensin peptides by mass spectrometric analysis. BGAP showed a broad spectrum of antifungal act  ...[more]

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