isolation-characterization-cyanobacterial-extracellular

Isolation and Characterization of Cyanobacterial Extracellular Vesicles

Cyanobacteria are a diverse group of photosynthetic, Gram-negative bacteria that play critical roles in global ecosystems and serve as essential ...

Universidade do Porto

In bacteria, the active transport of material from the interior to the exterior of the cell, or secretion, represents a very important mechanism of adaptation to the surrounding environment. The secretion of various types of biomolecules is mediated by a series of multiprotein complexes that cross the bacterial membrane(s), each complex dedicated to the secretion of specific substrates. In addition, biological material may also be released from the bacterial cell in the form of vesicles. Extracellular vesicles (EVs) are bilayered, nanoscale structures, derived from the bacterial cell envelope, which contain membrane components as well as soluble products. In cyanobacteria, the knowledge regarding EVs is lagging far behind compared to what is known about, for example, other Gram-negative bacteria. Here, we present a summary of the most important findings regarding EVs in Gram-negative bacteria, discussing aspects of their composition, formation processes and biological roles, and highlighting a number of technological applications tested. This lays the groundwork to raise awareness that the release of EVs by cyanobacteria likely represents an important, and yet highly disregarded, survival strategy. Furthermore, we hope to motivate future studies that can further elucidate the role of EVs in cyanobacterial cell biology and physiology.

Extracellular Vesicles: An Overlooked Secretion System in Cyanobacteria.

Hospital-acquired infections are still a major concern worldwide, being frequently related to bacterial biofilm formation on medical devices, and thus difficult to eradicate with conventional antimicrobial treatments. Therefore, infection-preventive solutions based on natural polymers are being investigated. Recently, a marine cyanobacterium-derived polymeric coating (CyanoCoating) has demonstrated great anti-adhesive potential when immobilized onto gold model substrates. In this work, we took this technology a step closer to an industrial application by covalently immobilizing CyanoCoating onto medical grade polyurethane (PU). This immobilization was developed through the introduction of linkable moieties onto a PU inert surface using different pre-treatments. Besides the application of the polydopamine (pDA) linker layer, other processes frequently found in industrial settings, such as atmospheric plasma (using O2 or N2 as reactive gases) and ozone surface activations, were evaluated. From all the pre-treatments tested, the ozone activation was the most promising since the obtained coating not only revealed a homogeneous distribution, but also significantly reduced the adhesion of two relevant etiological bacteria in static conditions (the Gram-positive Staphylococcus aureus and the Gram-negative Escherichia coli). Moreover, it also impaired E. coli biofilm formation under simulated urinary tract dynamic conditions, reinforcing the potential of CyanoCoating as an antibiotic-free alternative to mitigate medical device-associated infections, particularly in the urinary tract.

Surface activation of medical grade polyurethane for the covalent immobilization of an anti-adhesive biopolymeric coating.

Cyanobacteria are a group of photosynthetic prokaryotes that contribute to primary production on a global scale. These microorganisms release vesicles to the extracellular environment, spherical nanosized structures, derived essentially from the outer membrane. Even though earlier works in model Gram-negative bacteria have hypothesized that outer membrane stability is crucial in vesicle formation, the mechanisms determining vesicle biogenesis in cyanobacteria remain unknown. Here, we report on the identification of six candidate genes encoding outer membrane proteins harboring SLH/OprB-domains in the genome of the model cyanobacterium  sp. PCC 6803. Using a genetics-based approach, one gene was found to encode an essential protein (Slr1841), while the remaining five are not essential for growth under standard conditions. Vesicle production was monitored, and it was found that a mutant in the gene encoding the second most abundant SLH/OprB protein in  sp. PCC 6803 outer membrane (Slr1908) produces more vesicles than any of the other tested strains. Moreover, the Slr1908-protein was also found to be important for iron uptake. Altogether, our results suggest that proteins containing the SLH/OprB-domains may have dual biological role, related to micronutrient uptake and to outer membrane stability, which, together or alone, seem to be involved in cyanobacterial vesicle biogenesis.

Article	Year
Extracellular Vesicles: An Overlooked Secretion System in Cyanobacteria. Life (Basel, Switzerland)\| PubMed ID: 32751844	2020
Surface activation of medical grade polyurethane for the covalent immobilization of an anti-adhesive biopolymeric coating. Journal of materials chemistry. B\| PubMed ID: 33871523	2021
The Role of Outer Membrane Protein(s) Harboring SLH/OprB-Domains in Extracellular Vesicles' Production in sp. PCC 6803. Plants (Basel, Switzerland)\| PubMed ID: 34961227	2021

Jorge Matinha-Cardoso

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