PDF(Sci-hub)
Abstract：
Willaertia magna C2c Maky is a free-living amoeba that has demonstrated its ability to inhibit the intracellular multiplication of some Legionella pneumophila strains, which are pathogenic bacteria inhabiting the aquatic environment. The Amoeba, an industry involved in the treatment of microbiological risk in the water and plant protection sectors, has developed a natural biocide based on the property of W. magna to manage the proliferation of the pathogen in cooling towers. In axenic liquid medium, amoebas are usually cultivated in adhesion on culture flask. However, we implemented a liquid culture in suspension using bioreactors in order to produce large quantities of W. magna. In order to investigate the culture condition effects on W. magna, we conducted a study based on microscopic, proteomics and lipidomics analyzes. According to the culture condition, amoeba exhibited two different phenotypes. The differential proteomics study showed that amoebas seemed to promote the lipid metabolism pathway in suspension culture, whereas we observed an upregulation of the carbohydrate pathway in adherent culture. Furthermore, we observed an over-regulation of proteins related to the cytoskeleton for W. magna cells grown in adhesion. Regarding the lipid analysis, suspension and adhesion cell growth showed comparable lipid class compositions. However, the differential lipid analysis revealed differences that confirmed cell phenotype differences observed by microscopy and predicted by proteomics. Overall, this study provides us with a better insight into the biology and molecular processes of W. magna in different culture lifestyles.
摘要：
WillaertiamagnaC2cMaky是一种自由生活的变形虫，已证明其能够抑制某些嗜肺军团菌菌株的细胞内繁殖，是居住在水生环境中的致病菌。变形虫,在水和植物保护部门参与处理微生物风险的行业，已开发出一种基于W.magna特性的天然杀菌剂，以管理冷却塔中病原体的扩散。在无菌液体培养基中，变形虫通常在培养瓶上粘附培养。然而,为了生产大量的W.magna，我们使用生物反应器在悬浮液中进行液体培养。为了研究培养条件对W.magna的影响，我们进行了一项基于微观的研究，蛋白质组学和脂质组学分析。根据培养条件,变形虫表现出两种不同的表型。差异蛋白质组学研究表明，阿米巴似乎促进了悬浮培养中的脂质代谢途径，而我们观察到粘附培养中碳水化合物途径的上调。此外，我们观察到粘附生长的W.magna细胞的细胞骨架相关蛋白的过度调节。关于脂质分析，悬浮和粘附细胞生长显示出相当的脂质类组成。然而,差异脂质分析显示差异证实了显微镜观察和蛋白质组学预测的细胞表型差异。总的来说,这项研究为我们提供了更好的了解W.magna在不同文化生活方式中的生物学和分子过程。
