Abstract
A novel antifungal protein, Mr = ca. 40 kDa, was isolated from pumpkin rind and designated Pr-1. When purified by anion exchange chromatography and HPLC, it inhibited growth of several fungi including Botrytis cinerea, Fusarium oxysporum, Fusarium solani and Rhizoctonia solani, as well as the yeast, Candida albicans, at 10–20 μM. It did not inhibit growth of Escherichia coli or Staphylococcus aureus even at 200 μM. Laser scanning microscopy of fungal cells exposed to rhodamine-labeled Pr-1 revealed that the protein accumulated and was localized on the cell surface. Uptake of the vital stain, SYTOX Green, was enhanced when fungal conidia were treated with Pr-1 suggesting that the protein has membrane permeabilization activity. Pr-1 was thermostable at 70°C and did not lyse human red blood cells at 128 μM suggesting that the protein may be useful as an antifungal agent with little, if any human cytotoxicity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barbieri L, Polito L, Bolognesi A, Ciani M, Pelosi E, Farini V, Jha AK, Sharma N, Vivanco JM, Chambery A, Parente A, Stirpe F (2006) Ribosome-inactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata. Biochim Biophys Acta 1760:783–792
Cheong NE, Choi YO, Kim WY, Bae IS, Cho MJ, Hwang I, Kim JW, Lee SY (1997) Purification and characterization of an antifungal PR-5 protein from pumpkin leaves. Mol Cells 7:214–219
Fritig B, Heitz T, Legrand M (1998) Antimicrobial proteins in induced plant defense. Curr Opin Immunol 10:16–22
Gomes VM, Carvalho AO, Da Cunha M, Keller MN, Bloch C Jr, Deolindo P, Alves EW (2005) Purification and characterization of a novel peptide with antifungal activity from Bothrops jararaca venom. Toxicon 45:817–827
Kaomek M, Mizuno K, Fujimura T, Sriyotha P, Cairns JR (2003) Cloning, expression, and characterization of an antifungal chitinase from Leucaena leucocephala de Wit. Biosci Biotechnol Biochem 67:667–676
Kim MH, Park SC, Kim JY, Lee SY, Lim HT, Cheong H, Hahm KS, Park Y (2006) Purification and characterization of a heat-stable serine protease inhibitor from the tubers of new potato variety “Golden Valley”. Biochem Biophys Res Commun 346:681–686
Ng TB (2004) Antifungal proteins and peptides of leguminous and non-leguminous origins. Peptides 25:1215–1222
Ng TB, Parkash A (2002) Hispin, a novel ribosome inactivating protein with antifungal activity from hairy melon seeds. Protein Expr Purif 26:211–217
Park SC, Lee JR, Shin SO, Jung JH, Lee YM, Son H, Park Y, Lee SY, Hahm KS (2007a) Purification and characterization of an antifungal protein, C-FKBP, from Chinese cabbage. J Agric Food Chem 55:5277–5281
Park Y, Park SC, Park HK, Shin SY, Kim Y, Hahm KS (2007b) Structure–activity relationship of HP (2-20) analog peptide: enhanced antimicrobial activity by N-terminal random coil region deletion. Biopolymers 88:199–207
Park SC, Lee JR, Kim JY, Lee SS, Park Y, Cheong GW, Lee SY, Hahm KS (2007c) Molecular and functional characterization of a cyclophilin with antifungal activity from Chinese cabbage. Biochem Biophys Res Commun 353:672–678
Park SC, Lee JR, Shin SO, Park Y, Lee SY, Hahm KS (2007d) Characterization of a heat-stable protein with antimicrobial activity from Arabidopsis thaliana. Biochem Biophys Res Commun 362:562–567
Park SC, Kim MH, Hossain MA, Shin SY, Kim Y, Stella L, Wade JD, Park Y, Hahm KS (2008) Amphipathic alpha-helical peptide, HP (2-20), and its analogues derived from Helicobacter pylori: pore formation mechanism in various lipid compositions. Biochim Biophys Acta 1778:229–241
Sawano Y, Miyakawa T, Yamazaki H, Tanokura M, Hatano K (2007) Purification, characterization, and molecular gene cloning of an antifungal protein from Ginkgo biloba seeds. Biol Chem 388:273–280
Vassiliou AG, Neumann GM, Condron R, Polya GM (1998) Purification and mass spectrometry-assisted sequencing of basic antifungal proteins from seeds of pumpkin (Cucurbita maxima). Plant Sci 134:141–162
Wang HX, Ng TB (2003) Isolation of cucurmoschin, a novel antifungal peptide abundant in arginine, glutamate and glycine residues from black pumpkin seeds. Peptides 24:969–972
Ye XY, Wang H, Ng TB (1999) First chromatographic isolation of an antifungal thaumatin-like protein from French bean legumes and demonstration of its antifungal activity. Biochem Biophys Res Commun 263:130–134
Yi SY, Hwang BK (1997) Purification and antifungal activity of a basic 34 kDa beta-1, 3-glucanase from soybean hypocotyls inoculated with Phytophthora sojae f. sp. glycines. Mol Cells 7:408–413
Acknowledgement
This work was supported by the Korean Research Foundation Grant funded by the Korean Government [KRF-2008-359-C00022].
Author information
Authors and Affiliations
Corresponding authors
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Park, SC., Lee, J.R., Kim, JY. et al. Pr-1, a novel antifungal protein from pumpkin rinds. Biotechnol Lett 32, 125–130 (2010). https://doi.org/10.1007/s10529-009-0126-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-009-0126-y