Anti-Candida Albicans Effect of the Protein-carbohydrate Fraction Obtained from the Coelomic Fluid of Earthworm Dendrobaena Veneta

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2019 Mar 12

PMID 30856188

Citations 15

Authors

Marta J Fiolka

Paulina Czaplewska

Katarzyna Macur

Tomasz Buchwald

Jolanta Kutkowska

Roman Paduch

Zbigniew Kaczynski

Jerzy Wydrych

Teresa Urbanik-Sypniewska

Affiliations

Soon will be listed here.

Abstract

An antifungal active fraction (AAF) from the coelomic fluid (CF) of the earthworm Dendrobaena veneta was isolated. The aim of the study was to analyze the antifungal activity of the AAF and to carry out chemical characterization of the fraction. The active fraction showed antifungal activity against a clinical C. albicans isolate, C. albicans ATCC 10231, and C. krusei ATCC 6258. It effectively reduced the metabolic activity of C. albicans cells and influenced their morphology after 48 hours of incubation. Scanning electron microscopy (SEM) images revealed loss of integrity of the cell wall induced by the active fraction. Calcofluor White staining showed changes in the structure of the C. albicans cell wall induced by the AAF. The fungal cells died via apoptosis and necrosis after the treatment with the studied fraction. Electrophoresis under native conditions revealed the presence of two compounds in the AAF, while SDS/PAGE gel electrophoresis showed several protein and carbohydrate compounds. The active fraction was analyzed using Raman spectroscopy, MALDI TOF/TOF, and ESI LC-MS. The Raman analysis confirmed the presence of proteins and determined their secondary structure. The MALDI TOF/TOF analysis facilitated detection of four main compounds with a mass of 7694.9 m/z, 12292.3 m/z, 21628.3 m/z, and 42923.2 m/z in the analyzed fraction. The presence of carbohydrate compounds in the preparation was confirmed by nuclear magnetic resonance (NMR) and gas chromatography (GC-MS). The ATR-FTIR spectrum of the AAF exhibited high similarity to the spectrum of egg white lysozyme. The AAF showed no endotoxicity and cytotoxicity towards normal skin fibroblasts (HSF); therefore, it can be used for the treatment of skin and mucous membrane candidiasis in the future. Given its efficient and selective action, the fraction seems to be a promising preparation with antifungal activity against C. albicans.

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References

Sardi J, Scorzoni L, Bernardi T, Fusco-Almeida A, Mendes Giannini M . Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2012; 62(Pt 1):10-24. DOI: 10.1099/jmm.0.045054-0. View

Ibrahim H, Yamada M, Matsushita K, Kobayashi K, Kato A . Enhanced bactericidal action of lysozyme to Escherichia coli by inserting a hydrophobic pentapeptide into its C terminus. J Biol Chem. 1994; 269(7):5059-63. View

Mereghetti P, Corsetto P, Cremona A, Rizzo A, Doglia S, Ami D . A Fourier transform infrared spectroscopy study of cell membrane domain modifications induced by docosahexaenoic acid. Biochim Biophys Acta. 2014; 1840(10):3115-22. DOI: 10.1016/j.bbagen.2014.07.003. View

Liu Y, Sun Z, Wang C, Li S, Liu Y . Purification of a novel antibacterial short peptide in earthworm Eisenia foetida. Acta Biochim Biophys Sin (Shanghai). 2004; 36(4):297-302. DOI: 10.1093/abbs/36.4.297. View

Mukaremera L, Lee K, Mora-Montes H, Gow N . Yeast, Pseudohyphal, and Hyphal Morphogenesis Differentially Affects Immune Recognition. Front Immunol. 2017; 8:629. PMC: 5461353. DOI: 10.3389/fimmu.2017.00629. View

Hua Z, Wang Y, Cao H, Pu L, Cui Y . Purification of a protein from coelomic fluid of the earthworm Eisenia foetida and evaluation of its hemolytic, antibacterial, and antitumor activities. Pharm Biol. 2011; 49(3):269-75. DOI: 10.3109/13880209.2010.508498. View

Kim J, Sudbery P . Candida albicans, a major human fungal pathogen. J Microbiol. 2011; 49(2):171-7. DOI: 10.1007/s12275-011-1064-7. View

Sebaa S, Hizette N, Boucherit-Otmani Z, Courtois P . Dose‑dependent effect of lysozyme upon Candida albicans biofilm. Mol Med Rep. 2017; 15(3):1135-1142. PMC: 5367326. DOI: 10.3892/mmr.2017.6148. View

Monheit J, Cowan D, Moore D . Rapid detection of fungi in tissues using calcofluor white and fluorescence microscopy. Arch Pathol Lab Med. 1984; 108(8):616-8. View

10.

De Colibus L, Sonnen A, Morris K, Siebert C, Abrusci P, Plitzko J . Structures of lysenin reveal a shared evolutionary origin for pore-forming proteins and its mode of sphingomyelin recognition. Structure. 2012; 20(9):1498-507. PMC: 3526787. DOI: 10.1016/j.str.2012.06.011. View

11.

Lewandowska A, Macur K, Czaplewska P, Liss J, Lukaszuk K, Oldziej S . Qualitative and Quantitative Analysis of Proteome and Peptidome of Human Follicular Fluid Using Multiple Samples from Single Donor with LC-MS and SWATH Methodology. J Proteome Res. 2017; 16(8):3053-3067. DOI: 10.1021/acs.jproteome.7b00366. View

12.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

13.

Reisfeld R, Lewis U, Williams D . Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962; 195:281-3. DOI: 10.1038/195281a0. View

14.

Kozielski M, Buchwald T, Szybowicz M, Blaszczak Z, Piotrowski A, Ciesielczyk B . Determination of composition and structure of spongy bone tissue in human head of femur by Raman spectral mapping. J Mater Sci Mater Med. 2011; 22(7):1653-61. PMC: 3127018. DOI: 10.1007/s10856-011-4353-0. View

15.

Mazzoni C, Zarov P, RAMBOURG A, Mann C . The SLT2 (MPK1) MAP kinase homolog is involved in polarized cell growth in Saccharomyces cerevisiae. J Cell Biol. 1993; 123(6 Pt 2):1821-33. PMC: 2290862. DOI: 10.1083/jcb.123.6.1821. View

16.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

17.

Lefevre T, Paquet-Mercier F, Rioux-Dube J, Pezolet M . Review structure of silk by raman spectromicroscopy: from the spinning glands to the fibers. Biopolymers. 2011; 97(6):322-36. DOI: 10.1002/bip.21712. View

18.

Fiolka M, Grzywnowicz K, Chlebiej K, Szczuka E, Mendyk E, Keller R . Anti-Candida albicans action of the glyco-protein complex purified from metabolites of gut bacterium Raoultella ornithinolytica isolated from earthworms Dendrobaena veneta. J Appl Microbiol. 2012; 113(5):1106-19. DOI: 10.1111/j.1365-2672.2012.05407.x. View

19.

Fiolka M, Zagaja M, Hulas-Stasiak M, Wielbo J . Activity and immunodetection of lysozyme in earthworm Dendrobaena veneta (Annelida). J Invertebr Pathol. 2011; 109(1):83-90. DOI: 10.1016/j.jip.2011.10.002. View

20.

Zhang R, Wu L, Eckert T, Burg-Roderfeld M, Rojas-Macias M, Lutteke T . Lysozyme's lectin-like characteristics facilitates its immune defense function. Q Rev Biophys. 2017; 50:e9. DOI: 10.1017/S0033583517000075. View