Increased production of outer membrane vesicles by cultured freshwater bacteria in response to ultraviolet radiation

Silva, Thiago Pereira da; Dias, Felipe F.; Melo, Rossana C. N.

doi:10.1016/j.micres.2016.08.002

Cited by 35 publications

(44 citation statements)

References 42 publications

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“…3k ) might indicate that phages themselves are released inside MVs. Moreover, UV radiation of water samples, a trigger for prophage induction, was demonstrated to greatly stimulate vesicle formation 42 .…”

Section: Discussionmentioning

confidence: 99%

Prophage-triggered membrane vesicle formation through peptidoglycan damage in Bacillus subtilis

et al. 2017

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Bacteria release membrane vesicles (MVs) that play important roles in various biological processes. However, the mechanisms of MV formation in Gram-positive bacteria are unclear, as these cells possess a single cytoplasmic membrane that is surrounded by a thick cell wall. Here we use live cell imaging and electron cryo-tomography to describe a mechanism for MV formation in Bacillus subtilis. We show that the expression of a prophage-encoded endolysin in a sub-population of cells generates holes in the peptidoglycan cell wall. Through these openings, cytoplasmic membrane material protrudes into the extracellular space and is released as MVs. Due to the loss of membrane integrity, the induced cells eventually die. The vesicle-producing cells induce MV formation in neighboring cells by the enzymatic action of the released endolysin. Our results support the idea that endolysins may be important for MV formation in bacteria, and this mechanism may potentially be useful for the production of MVs for applications in biomedicine and nanotechnology.

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Section: Discussionmentioning

confidence: 99%

Prophage-triggered membrane vesicle formation through peptidoglycan damage in Bacillus subtilis

et al. 2017

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“…Reports indicate that different stress conditions, including antibiotics (28), temperature (10), ultraviolet irradiation (28, 33), virulence factors (34), SOS response (35), oxygen stress (36), nutritional deprivation (28) and physical pressure (34), can affect the number of vesicles released from bacteria. In this study, the release of OMVs was stimulated by adding heavy metals to the environment when the bacteria growth rate was unrestricted.…”

Section: Discussionmentioning

confidence: 99%

Outer Membrane Vesicles Mediated Horizontal Transfer of an Aerobic Denitrification Gene betweenEscherichia coli

Luo

Miao

Qiao

2019

Preprint

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16Bacterial genetic material can be horizontally transferred between microorganisms 17 via outer membrane vesicles (OMVs) released by bacteria. Up to now, the application of 18 vesicle-mediated horizontal transfer of "degrading genes" in environmental remediation 19 has not been reported. In this study, the nirS gene from an aerobic denitrification 20 bacterium, Pseudomonas stutzeri, was enclosed in a pET28a plasmid, transformed into 21 Escherichia coli (E. coli) DH5α and expressed in E. coli BL21. The E. coli DH5α 22 released OMVs containing the recombination plasmid pET28a-nirS. Moreover, the 23 amount of released OMVs-protein and DNA in OMVs increase as heavy metal 24 concentrations and temperature increased. When compared with the free pET28a-nirS 25 plasmid's inability to transform, nirS in OMVs could be transferred into E. coli BL21 26 with the transformation frequency of 2.76×10 6 CFU/g when the dosage of OMVs was 27 200 µg under natural conditions, and nirS could express successfully in recipient bacteria. 28 Furthermore, the recipient bacteria that received OMVs could produce 18.16 U ml -1 29 activity of nitrite reductase. Vesicle-mediated HGT of aerobic denitrification genes 30 provides a novel bioaugmentation technology of nitrogen removal. 31 32 Importance 33Previous studies have reported that bacterial genetic material can be horizontally 34 transferred between microorganisms via outer membrane vesicles(OMVs) released by 35 bacteria. However, the application of vesicle-mediated horizontal transfer of "degrading 36 genes" in environmental remediation has not been reported. In this study, we found that 37 OMVs could mediate horizontal transfer of pET28a-nirS plasmid between E. coli under 38 3 natural condition. The transformation frequency reached to 2.76×10 6 , which was higher 39 than that of the free plasmid. Vesicle-mediated HGT of aerobic denitrification genes 40 provides a novel bioaugmentation technology of nitrogen removal. 41 42 44 45 4 Introduction 46High nitrogen concentrations in water result in water eutrophication and pollution. 47Traditional bio-treatment processes for nitrogen removal involve autotrophic and 48 heterotrophic denitrification under aerobic and anoxic conditions, respectively (1). 49Because of their different oxygen requirements, these two steps are separated spatially 50 and temporally. Recently, more researchers have focused on nitrogen removal using 51 aerobic denitrification bacteria. Unlike traditional anaerobic denitrification mechanisms, 52 aerobic denitrification occurs via co-respiration or co-metabolism of O 2 and NO 3 -. 53Additionally, nitrification and denitrification can occur in the same aerobic denitrification 54 system (2, 3). Nitrate reductase (napA), nitrite reductase (nirS), nitric oxide reductase 55 (norB) and nitrous oxide reductase (nosZ) are four key enzymes in the aerobic 56 denitrification process of aerobic denitrifying bacteria. The aerobic denitrification 57 bacteria or microbial consortium can be added into wastewater for promoting the remo...

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“…The identification of specific protein components within OMVs may have great value for understanding the mechanism by which the OMVs interact with and regulate the innate immune responses in the host. Isolation of bacterial OMVs from culture medium has been reported [9][10][11], but bacterial OMVs from fecal samples may provide better insight into the functioning of the gut microbial community. Herein, we present an integrated methodology for enriching OMVs from human and mouse stool samples for identification of proteins carried in these bacterial OMVs.…”

Section: Introductionmentioning

confidence: 99%

A Method for Isolation and Proteomic Analysis of Outer Membrane Vesicles from Fecal Samples by LC-MS/MS

Zhu

et al. 2019

J Proteomics Bioinform

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Outer membrane vesicles (OMVs) are nanosized spheres secreted by bacteria that are similar to the vesicles known as exosomes, which are secreted by most mammalian cell types. In contrast to many studies focusing on optimizing methods for enriching exosomes from biological fluid, few studies have been conducted to investigate outer membrane vesicles from fecal samples. Herein, we have developed a pipeline comprised of membrane filtration and multiple cycles of ultracentrifugation (UC) to isolate OMVs from fecal samples for proteomics analysis, where multiple cycles of UC are required for removal of contaminants. By iTRAQ labeling quantitative proteomics analysis, different filter sizes (0.22 µm and 0.45 µm) were compared in terms of their performance in enriching OMVs and eliminating background fecal material. Using the 0.45 µm filter, a slightly higher protein yield was obtained but no additional contaminating proteins from bacteria were identified compared to those from the 0.22 µm filter. The 0.45 µm filter together with the multiple cycles of UC were thus used to isolate OMVs for proteomics analysis. To our knowledge, this is the first study profiling a large number of OMV proteins from fecal samples. Such capabilities may help provide valuable information in understanding the communication between the host and microbiota, which is critical in preventing cancer and disease development..

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Increased production of outer membrane vesicles by cultured freshwater bacteria in response to ultraviolet radiation

Cited by 35 publications

References 42 publications

Prophage-triggered membrane vesicle formation through peptidoglycan damage in Bacillus subtilis

Prophage-triggered membrane vesicle formation through peptidoglycan damage in Bacillus subtilis

Outer Membrane Vesicles Mediated Horizontal Transfer of an Aerobic Denitrification Gene betweenEscherichia coli

A Method for Isolation and Proteomic Analysis of Outer Membrane Vesicles from Fecal Samples by LC-MS/MS

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