Anti-Candida albicans effect of the protein-carbohydrate fraction obtained from the coelomic fluid of earthworm Dendrobaena veneta

Marta J. Fiołka , Paulina Czaplewska , Katarzyna Macur , Tomasz Buchwald , Jolanta Kutkowska , Roman Paduch , Zbigniew Kaczyński , Jerzy Wydrych , Teresa Urbanik-Sypniewska

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.
Author Marta J. Fiołka
Marta J. Fiołka,,
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, Paulina Czaplewska (IFB / ZLS)
Paulina Czaplewska,,
- Zespół Laboratoriów Specjalistycznych MWB UG i GUMed
, Katarzyna Macur (IFB / ZLS)
Katarzyna Macur,,
- Zespół Laboratoriów Specjalistycznych MWB UG i GUMed
, Tomasz Buchwald
Tomasz Buchwald,,
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, Jolanta Kutkowska
Jolanta Kutkowska,,
-
, Roman Paduch
Roman Paduch,,
-
, Zbigniew Kaczyński (FCh / DBCh / LSB)
Zbigniew Kaczyński,,
- Laboratory of Structural Biochemistry
, Jerzy Wydrych
Jerzy Wydrych,,
-
, Teresa Urbanik-Sypniewska
Teresa Urbanik-Sypniewska,,
-
Journal seriesPlos One, ISSN 1932-6203, (A 35 pkt)
Issue year2019
Vol14
No3
Pages1-30
Publication size in sheets1.45
Article numbere0212869
ASJC Classification1100 General Agricultural and Biological Sciences; 1300 General Biochemistry, Genetics and Molecular Biology; 2700 General Medicine
DOIDOI:10.1371/journal.pone.0212869
URL https://doi.org/10.1371/journal.pone.0212869
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)35
ScoreMinisterial score = 35.0, 24-07-2019, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2017 = 1.111; WoS Impact Factor: 2017 = 2.766 (2) - 2017=3.352 (5)
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