Department of Immunobiology, Institute of Biology and Biochemistry, Maria Curie-Skłodowska University, Lublin, Poland.
J Appl Microbiol. 2012 Nov;113(5):1106-19. doi: 10.1111/j.1365-2672.2012.05407.x. Epub 2012 Aug 21.
The aim of our research was to isolate the compounds from the metabolites of Raoultella ornithinolytica with the activity against Candida albicans and to analyse the action of the compounds on the metabolic activity and morphology of the fungus cells.
The effect of active protein fractions on the cell morphology, growth, and metabolic activity of C. albicans was analysed under a light microscope with Nomarski contrast and after staining with calcofluor-white. The LIVE/DEAD Yeast Viability Kit F-7030 FUN 1 was used for determination of C. albicans metabolic activity. The biomolecules obtained after isolation by ion exchange chromatography were further fractionated by Sephadex G-50 medium gel filtration. Then, after molecular sieve, the fractions were analysed by FTIR and SERS Spectroscopy. A subfraction was isolated from the antifungal protein fraction above 100 kDa. The active subfraction identified as the glyco-protein complex caused a decrease in the metabolic activity and morphological changes of C. albicans cells.
The glyco-protein complex obtained from metabolites of bacteria Raoultella ornithinolytica possesses antifungal activity against C. albicans and shows minimal toxicity (1%) against fibroblasts.
Studies on the glyco-protein complex obtained from earthworm gut bacteria R. ornithinolytica can lead to their application in biological fungicide and pharmaceutical industry.
我们的研究目的是从鸟氨酸罗伊氏菌的代谢产物中分离出具有抗白色念珠菌活性的化合物,并分析这些化合物对真菌细胞代谢活性和形态的作用。
在相差显微镜下,用钙荧光白染色,分析活性蛋白部分对白色念珠菌细胞形态、生长和代谢活性的影响。使用 LIVE/DEAD 酵母活力试剂盒 F-7030 FUN 1 测定白色念珠菌的代谢活性。通过离子交换层析分离得到的生物分子,进一步用 Sephadex G-50 中凝胶过滤进行分级。然后,经过分子筛,用 FTIR 和 SERS 光谱分析各馏分。从 100 kDa 以上的抗真菌蛋白部分分离出一个亚馏分。鉴定为糖蛋白复合物的活性亚馏分,导致白色念珠菌细胞代谢活性下降和形态发生变化。
从鸟氨酸罗伊氏菌代谢产物中获得的糖蛋白复合物对白色念珠菌具有抗真菌活性,对成纤维细胞的毒性最小(1%)。
对从蚯蚓肠道细菌罗伊氏菌中获得的糖蛋白复合物的研究,可能导致其在生物杀菌剂和制药工业中的应用。