Bacterial membrane vesicle functions, laboratory methods, and applications

Çelik, PınarAytar; Derkuş, Burak; Erdoğan, Kübra; Barut, Dilan; Enuh, Blaise Manga; Yıldırım, Yalın; Pecha, Simon; Çabuk, Ahmet

doi:10.1016/j.biotechadv.2021.107869

Cited by 39 publications

(67 citation statements)

References 220 publications

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“…Only 4% of the studies on prokaryotic membrane vesicles conducted between 1972 and 2021 concern MVs of Gram-positive bacteria 17 : indeed, it has been assumed for a long time that Gram-positive bacteria could not produce vesicles, because of the lack of an outer membrane layer and the presence of thick cell walls 1 . In recent years, Gram-positive MVs have gained more attention and their release was demonstrated in both pathogenic and non-pathogenic strains, such as Staphylococcus spp., Bacillus spp., Lactobacillus spp., and Streptomyces spp.…”

Section: Introductionmentioning

confidence: 99%

Unravelling the DNA sequences carried by Streptomyces coelicolor membrane vesicles

Faddetta

Vassallo

Duca

et al. 2022

Sci Rep

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Membrane vesicles (MVs) are spherical particles with nanoscale dimensions and characterized by the presence of diverse cargos, such as nucleic acids, proteins, lipids, and cellular metabolites. Many examples of (micro)organisms producing MVs are reported in literature. Among them, bacterial MVs are of particular interest because they are now considered as the fourth mechanism of horizontal gene transfer. Streptomyces bacteria are well-known for their ecological roles and ability to synthesize bioactive compounds, with Streptomyces coelicolor being the model organism. It was previously demonstrated that it can produce distinct populations of MVs characterized by different protein and metabolite cargos. In this work we demonstrated for the first time that MVs of S. coelicolor carry both DNA and RNA and that their DNA content represents the entire chromosome of the bacterium. These findings suggest that MV DNA could have a role in the evolution of Streptomyces genomes and that MVs could be exploited in new strain engineering strategies.

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Section: Introductionmentioning

confidence: 99%

Unravelling the DNA sequences carried by Streptomyces coelicolor membrane vesicles

Faddetta

Vassallo

Duca

et al. 2022

Sci Rep

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“…However, we experienced some unavoidable limitations across this study. Although the BCA assay is commonly used method to quantify MVs (Aytar Celik et al 2022 ; Bitto et al 2021 ), using this method to unify the concentration of two kinds of MVs solutions may have some influence on the experimental results because the protein content of the single MV in S. mutans and S. mutans Δ gtfBC mutant may be different. Furthermore, both in vitro and in vivo interactions of S. mutans MVs with mixed biofilms of S. gordonii and S. sanguinis is unclear.…”

Section: Discussionmentioning

confidence: 99%

Streptococcus mutans membrane vesicles inhibit the biofilm formation of Streptococcus gordonii and Streptococcus sanguinis

Cui

et al. 2022

AMB Expr

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Streptococcus mutans, whose main virulence factor is glucosyltransferase (Gtf), has a substantial impact on the development of dental caries. S. mutans membrane vesicles (MVs), which are rich in Gtfs, have been shown to affect biofilm formation of other microorganisms. Streptococcus gordonii and Streptococcus sanguinis are initial colonizers of tooth surfaces, which provide attachment sites for subsequent microorganisms and are crucial in the development of oral biofilms. S. mutans and S. gordonii, as well as S. mutans and S. sanguinis, have a complex competitive and cooperative relationship, but it is unclear whether S. mutans MVs play a role in these interspecific interactions. Therefore, we co-cultured S. mutans MVs, having or lacking Gtfs, with S. gordonii and S. sanguinis. Our results showed that S. mutans MVs inhibited biofilm formation of S. gordonii and S. sanguinis but did not affect their planktonic growth; contrastingly, S. mutans ΔgtfBC mutant MVs had little effect on both their growth and biofilm formation. Additionally, there were fewer and more dispersed bacteria in the biofilms of the S. mutans MV-treated group than that in the control group. Furthermore, the expression levels of the biofilm-related virulence factors GtfG, GtfP, and SpxB in S. gordonii and S. sanguinis were significantly downregulated in response to S. mutans MVs. In conclusion, the results of our study showed that S. mutans MVs inhibited biofilm formation of S. gordonii and S. sanguinis, revealing an important role for MVs in interspecific interactions.

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“…35,38 Moreover, inhibiting PG layer construction would disrupt the cell wall and alter the cell morphology, thus leading to outer membrane wrinkling and OMV formation. 33,39 The specific components of outer membrane are inportant for OMV formation. The exterior leaflet of lipopolysaccharide (LPS) is a key composition of the outer membrane structure, whose abundance in OMVs is higher than that in the outer membrane.…”

Section: Gene Target Identification and Modularmentioning

confidence: 99%

Engineering Outer Membrane Vesicles to Increase Extracellular Electron Transfer of Shewanella oneidensis

Lan

et al. 2023

ACS Synth. Biol.

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Outer membrane vesicles (OMVs) of Gram-negative bacteria play an essential role in cellular physiology. The underlying regulatory mechanism of OMV formation and its impact on extracellular electron transfer (EET) in the model exoelectrogenShewanella oneidensis MR-1 remain unclear and have not been reported. To explore the regulatory mechanism of OMV formation, we used the CRISPR-dCas9 gene repression technology to reduce the crosslink between the peptidoglycan (PG) layer and the outer membrane, thus promoting the OMV formation. We screened the target genes that were potentially beneficial to the outer membrane bulge, which were classified into two modules: PG integrity module (Module 1) and outer membrane component module (Module 2). We found that downregulation of the penicillin-binding protein-encoding gene pbpC for peptidoglycan integrity (Module 1) and the N-acetyl-d-mannosamine dehydrogenase-encoding gene wbpP involved in lipopolysaccharide synthesis (Module 2) exhibited the highest production of OMVs and enabled the highest output power density of 331.3 ± 1.2 and 363.8 ± 9.9 mW m–2, 6.33- and 6.96-fold higher than that of the wild-typeS. oneidensis MR-1 (52.3 ± 0.6 mW m–2), respectively. To elucidate the specific impacts of OMV formation on EET, OMVs were isolated and quantified for UV–visible spectroscopy and heme staining characterization. Our study showed that abundant outer membrane c-type cytochromes (c-Cyts) including MtrC and OmcA and periplasmic c-Cyts were exposed on the surface or inside of OMVs, which were the vital constituents responsible for EET. Meanwhile, we found that the overproduction of OMVs could facilitate biofilm formation and increase biofilm conductivity. To the best of our knowledge, this study is the first to explore the mechanism of OMV formation and its correlation with EET of S. oneidensis, which paves the way for further study of OMV-mediated EET.

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Bacterial membrane vesicle functions, laboratory methods, and applications

Cited by 39 publications

References 220 publications

Unravelling the DNA sequences carried by Streptomyces coelicolor membrane vesicles

Unravelling the DNA sequences carried by Streptomyces coelicolor membrane vesicles

Streptococcus mutans membrane vesicles inhibit the biofilm formation of Streptococcus gordonii and Streptococcus sanguinis

Engineering Outer Membrane Vesicles to Increase Extracellular Electron Transfer of Shewanella oneidensis

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