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Open AccessArticle

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

by Caicheng Wang 1,2,3, Yao Zhang 1,2,3, Weiwei Zhang 1,2,3, Susu Yuan 1,2,3, Tzibun Ng 4 and Xiujuan Ye 1,2,3,*
1
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China
2
Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China
3
Fujian Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
4
School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong 999077, China
*
Author to whom correspondence should be addressed.
Academic Editor: Marcello Locatelli
Molecules 2019, 24(7), 1337; https://doi.org/10.3390/molecules24071337
Received: 16 March 2019 / Revised: 2 April 2019 / Accepted: 2 April 2019 / Published: 4 April 2019
(This article belongs to the Special Issue Natural Product Pharmacology and Medicinal Chemistry)
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 μg/mL, 12.37 μg/mL, 16.81 μg/mL, and 5.60 μg/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. View Full-Text
Keywords: Brassica oleracea var. gongylodes; antifungal peptide; purification Brassica oleracea var. gongylodes; antifungal peptide; purification
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Wang, C.; Zhang, Y.; Zhang, W.; Yuan, S.; Ng, T.; Ye, X. Purification of an Antifungal Peptide from Seeds of Brassica oleracea var. gongylodes and Investigation of Its Antifungal Activity and Mechanism of Action. Molecules 2019, 24, 1337.

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