Bacterial nanotubes and their role as bacterial nanowires in Pseudomonas aeruginosa biofilms

<p>Microbial nanowires are nanofilaments that could offer an extracellular electron transfer (EET) pathway linking the bacterial respiratory chain to external surfaces, such as oxidized metals in the environment and engineered electrodes in renewable energy devices. Filaments proposed to function as nanowires have been reported in multiple bacteria, yet it remains largely unclear about the composition and electron transfer mechanism of bacterial nanowires. Pseudomonas aeruginosa is an environmental and electrochemically active bacterium. In this study, we found nanotube-like extracellular filaments in P. aeruginosa biofilms, which were bacterial membrane extensions similar to the nanowires reported in Shewanella oneidensis. Remarkably, conductive probe atomic force microscope showed measurable conductivity of these extracellular filaments, suggesting that they may function as nanowires in P. aeruginosa. Our results also indicated that the electron shuttle pyocyanin significantly affected the conductivity of P. aeruginosa nanowires, suggesting that the electron transfer mechanism of P. aeruginosa nanowires was different from S. oneidensis. Furthermore, factors that impact biofilm formation, such as flagella, type IV pili, and exopolysaccharides, were not essential for nanowires formation, while affect the formation and length of nanowires of P. aeruginosa. Taken together, this is the first report that investigated the role of electron shuttle on the conductivity of nanowires and factors that affected nanowires formation.</p>

Structure of a cytochrome-based bacterial nanowire

Electrically conductive pili from Geobacter species, termed bacterial “nanowires”, are intensely studied for their biological significance and potential in the development of new materials. We have characterized a unique nanowire from conductive G. sulfurreducens pili preparations by cryo-electron microscopy composed solely of the c-type cytochrome OmcS. We present here, at 3.4 Å resolution, a novel structure of a cytochrome-based filament and discuss its possible role in long-range biological electron transport.Summary sentenceCryo-electron microscopy reveals the remarkable assembly of a c-type cytochrome into filaments comprising a heme-based bacterial nanowire.

Ultrastructure of Shewanella oneidensis MR-1 nanowires revealed by electron cryotomography

Bacterial nanowires have garnered recent interest as a proposed extracellular electron transfer (EET) pathway that links the bacterial electron transport chain to solid-phase electron acceptors away from the cell. Recent studies showed that Shewanella oneidensis MR-1 produces outer membrane (OM) and periplasmic extensions that contain EET components and hinted at their possible role as bacterial nanowires. However, their fine structure and distribution of cytochrome electron carriers under native conditions remained unclear, making it difficult to evaluate the potential electron transport (ET) mechanism along OM extensions. Here, we report high-resolution images of S. oneidensis OM extensions, using electron cryotomography (ECT). We developed a robust method for fluorescence light microscopy imaging of OM extension growth on electron microscopy grids and used correlative light and electron microscopy to identify and image the same structures by ECT. Our results reveal that S. oneidensis OM extensions are dynamic chains of interconnected outer membrane vesicles (OMVs) with variable dimensions, curvature, and extent of tubulation. Junction densities that potentially stabilize OMV chains are seen between neighboring vesicles in cryotomograms. By comparing wild type and a cytochrome gene deletion mutant, our ECT results provide the likely positions and packing of periplasmic and outer membrane proteins consistent with cytochromes. Based on the observed cytochrome packing density, we propose a plausible ET path along the OM extensions involving a combination of direct hopping and cytochrome diffusion. A mean-field calculation, informed by the observed ECT cytochrome density, supports this proposal by revealing ET rates on par with a fully packed cytochrome network.

Multifunctional microspherical magnetic and pH responsive carriers for combination anticancer therapy engineered by droplet-based microfluidics

Drug loaded luminescent porous silicon diatoms and magnetic bacterial nanowires were encapsulated within pH sensitive polymer forming biodegradable microcapsules using droplet-based microfluidics for targeting colorectal cancer.