Anti-Candida albicans effect of the protein-carbohydrate fraction obtained from the coelomic fluid of earthworm Dendrobaena veneta
Anti-Candida albicans effect of the protein-carbohydrate fraction obtained from the coelomic fluid of earthworm Dendrobaena veneta
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DOI：
10.1371/journal.pone.0212869
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发表时间：
2019-03-11
期刊：
影响因子：
3.7
通讯作者：
Urbanik-Sypniewska, Teresa
中科院分区：
文献类型：
--
作者：
Fiolka, Marta J.;Czaplewska, Paulina;Urbanik-Sypniewska, Teresa
从蚯蚓 Dendrobaena veneta 的体腔液 (CF) 中分离出抗真菌活性组分 (AAF)。该研究的目的是分析 AAF 的抗真菌活性并对该组分进行化学表征。该活性组分对临床分离的白色念珠菌、白色念珠菌 ATCC 10231 和克柔念珠菌 ATCC 6258 表现出抗真菌活性。它有效降低了白色念珠菌细胞的代谢活性，并在孵育 48 小时后影响其形态。扫描电子显微镜 (SEM) 图像显示活性组分导致细胞壁完整性丧失。 Calcoflor White 染色显示 AAF 诱导的白色念珠菌细胞壁结构发生变化。用所研究的级分处理后，真菌细胞通过凋亡和坏死而死亡。天然条件下的电泳显示 AAF 中存在两种化合物，而 SDS/PAGE 凝胶电泳显示多种蛋白质和碳水化合物化合物。使用拉曼光谱、MALDI TOF/TOF 和 ESI LC-MS 分析活性部分。拉曼分析证实了蛋白质的存在并确定了它们的二级结构。 MALDI TOF/TOF 分析有助于检测分析馏分中质量分别为 7694.9 m/z、12292.3 m/z、21628.3 m/z 和 42923.2 m/z 的四种主要化合物。通过核磁共振（NMR）和气相色谱（GC-MS）证实制剂中存在碳水化合物化合物。 AAF 的 ATR-FTIR 光谱与蛋清溶菌酶的光谱高度相似。 AAF对正常皮肤成纤维细胞（HSF）没有表现出内毒性和细胞毒性；因此，今后可用于治疗皮肤粘膜念珠菌病。鉴于其高效和选择性的作用，该级分似乎是一种有前途的制剂，具有针对白色念珠菌的抗真菌活性。
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.