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脂肽生物表面活性剂黏性素增强荧光假单胞菌SBW25生物膜的扩散。

Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms.

作者信息

Bonnichsen Lise, Bygvraa Svenningsen Nanna, Rybtke Morten, de Bruijn Irene, Raaijmakers Jos M, Tolker-Nielsen Tim, Nybroe Ole

机构信息

1​ Section for Microbial Ecology and Biotechnology, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.

2​ Costerton Biofilm Center, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

出版信息

Microbiology (Reading). 2015 Dec;161(12):2289-97. doi: 10.1099/mic.0.000191. Epub 2015 Sep 28.

Abstract

Pseudomonads produce several lipopeptide biosurfactants that have antimicrobial properties but that also facilitate surface motility and influence biofilm formation. Detailed studies addressing the significance of lipopeptides for biofilm formation and architecture are rare. Hence, the present study sets out to determine the specific role of the lipopeptide viscosin in Pseudomonas fluorescens SBW25 biofilm formation, architecture and dispersal, and to relate viscA gene expression to viscosin production and effect. Initially, we compared biofilm formation of SBW25 and the viscosin-deficient mutant strain SBW25ΔviscA in static microtitre assays. These experiments demonstrated that viscosin had little influence on the amount of biofilm formed by SBW25 during the early stages of biofilm development. Later, however, SBW25 formed significantly less biofilm than SBW25ΔviscA. The indication that viscosin is involved in biofilm dispersal was confirmed by chemical complementation of the mutant biofilm. Furthermore, a fluorescent bioreporter showed that viscA expression was induced in biofilms 4 h prior to dispersal. Subsequent detailed studies of biofilms formed in flow cells for up to 5 days revealed that SBW25 and SBW25ΔviscA developed comparable biofilms dominated by well-defined, mushroom-shaped structures. Carbon starvation was required to obtain biofilm dispersal in this system. Dispersal of SBW25 biofilms was significantly greater than of SBW25ΔviscA biofilms after 3 h and, importantly, carbon starvation strongly induced viscA expression, in particular for cells that were apparently leaving the biofilm. Thus, the present study points to a role for viscosin-facilitated motility in dispersal of SBW25 biofilms.

摘要

假单胞菌能产生多种具有抗菌特性的脂肽生物表面活性剂,这些生物表面活性剂还能促进表面运动并影响生物膜的形成。针对脂肽在生物膜形成和结构方面重要性的详细研究很少。因此,本研究旨在确定脂肽类毒素在荧光假单胞菌SBW25生物膜形成、结构和分散中的具体作用,并将viscA基因表达与类毒素的产生及作用联系起来。最初,我们在静态微孔试验中比较了SBW25和缺乏类毒素的突变菌株SBW25ΔviscA的生物膜形成情况。这些实验表明,在生物膜发育的早期阶段,类毒素对SBW25形成的生物膜量影响很小。然而,后来SBW25形成的生物膜明显少于SBW25ΔviscA。突变体生物膜的化学互补证实了类毒素参与生物膜分散的迹象。此外,一种荧光生物报告基因显示,在生物膜分散前4小时viscA表达被诱导。随后对流动小室中形成长达5天的生物膜进行的详细研究表明,SBW25和SBW25ΔviscA形成了类似的生物膜,其主要由明确的蘑菇状结构主导。在该系统中需要碳饥饿才能实现生物膜的分散。3小时后,SBW25生物膜的分散程度明显大于SBW25ΔviscA生物膜,重要的是,碳饥饿强烈诱导viscA表达,特别是对于那些明显离开生物膜的细胞。因此,本研究指出类毒素促进的运动在SBW25生物膜分散中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80b2/4811653/e4403e1d0041/mic000191-f1.jpg

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