Schrempf Hildgund, Merling Philipp
FB Biology/Chemistry, Applied Genetics of Microorganisms, University Osnabrück, Barbarastr. 13, D-49069, Osnabrück, Germany.
Microb Biotechnol. 2015 Jul;8(4):644-58. doi: 10.1111/1751-7915.12274. Epub 2015 Apr 7.
We selected Streptomyces lividans to elucidate firstly the biogenesis and antimicrobial activities of extracellular vesicles that a filamentous and highly differentiated Gram-positive bacterium produces. Vesicle types range in diameter from 110 to 230 nm and 20 to 60 nm, respectively; they assemble to clusters, and contain lipids and phospholipids allowing their in situ imaging by specific fluorescent dyes. The presence of the identified secondary metabolite undecylprodigiosin provokes red fluorescence of a portion of the heterogeneous vesicle populations facilitating in vivo monitoring. Protuberances containing vesicles generate at tips, and alongside of substrate hyphae, and enumerate during late vegetative growth to droplet-like exudates. Owing to in situ imaging in the presence and absence of a green fluorescent vancomycin derivative, we conclude that protuberances comprising vesicles arise at sites with enhanced levels of peptidoglycan subunits [pentapeptide of lipid II (C55)-linked disaccharides], and reduced levels of polymerized and cross-linked peptidoglycan within hyphae. These sites correlate with enhanced levels of anionic phospholipids and lipids. Vesicles provoke pronounced damages of Aspergillus proliferans, Verticillium dahliae and induced clumping and distortion of Escherichia coli. These harmful effects are likely attributable to the action of the identified vesicular compounds including different enzyme types, components of signal transduction cascades and undecylprodigiosin. Based on our pioneering findings, we highlight novel clues with environmental implications and application potential.
我们选择了变铅青链霉菌,首先来阐明一种丝状且高度分化的革兰氏阳性细菌所产生的细胞外囊泡的生物合成及抗菌活性。囊泡类型的直径分别为110至230纳米和20至60纳米;它们聚集成簇,并含有脂质和磷脂,这使得它们能够通过特定荧光染料进行原位成像。已鉴定出的次生代谢产物十一烷基灵菌红素的存在会引发一部分异质囊泡群体发出红色荧光,便于进行体内监测。含有囊泡的突起在顶端以及底物菌丝旁产生,并在营养生长后期形成液滴状渗出物。由于在存在和不存在绿色荧光万古霉素衍生物的情况下进行了原位成像,我们得出结论,包含囊泡的突起出现在肽聚糖亚基[脂质II(C55)连接的二糖的五肽]水平升高、菌丝内聚合和交联肽聚糖水平降低的部位。这些部位与阴离子磷脂和脂质水平的升高相关。囊泡对增殖曲霉、大丽轮枝菌造成了明显损害,并导致大肠杆菌聚集和变形。这些有害影响可能归因于已鉴定出的囊泡化合物的作用,包括不同类型的酶、信号转导级联的成分和十一烷基灵菌红素。基于我们的开创性发现,我们突出了具有环境意义和应用潜力的新线索。