Bacteria‐Based Production of Thiol‐Clickable, Genetically Encoded Lipid Nanovesicles

Abstract

Despite growing research efforts on the preparation of (bio)functional liposomes, synthetic capsules cannot reach the densities of protein loading and the control over peptide display that is achieved by natural vesicles. Herein, a microbial platform for high‐yield production of lipidic nanovesicles with clickable thiol moieties in their outer corona is reported. These nanovesicles show low size dispersity, are decorated with a dense, perfectly oriented, and customizable corona of transmembrane polypeptides. Furthermore, this approach enables encapsulation of soluble proteins into the nanovesicles. Due to the mild preparation and loading conditions (absence of organic solvents, pH gradients, or detergents) and their straightforward surface functionalization, which takes advantage of the diversity of commercially available maleimide derivatives, bacteria‐based proteoliposomes are an attractive eco‐friendly alternative that can outperform currently used liposomes.

Stable anchoring of bacteria-based protein nanoparticles for surface enhanced cell guidance

“…GFP IB-like pNPs produced by E. coli and L. lactis were covalently attached to maleimide-terminated SAMs through their cysteine aminoacids by means of a Michael addition interfacial reaction using the mCP technique, as recently shown for other nano-objects. [46][47][48] More specifically, the exposed thiol groups of the cysteine residues of GFP at the surface of the pNPs reacted with the maleimide groups of the SAM forming covalent bonds, thus providing a more stable binding in comparison with a simple physisorption procedure (Fig. 1).…”

Red Blood Cell Extracellular Vesicle-Based Drug Delivery: Challenges and Opportunities

“…The types of natural lipid NVs include exosomes, virosomes, bacterial ghosts, and erythrocyte ghosts (60). Conversely, synthetic NVs were created to mimic the physicochemical properties of liposomes (61). Liposomes contain a lipid bilayer surrounding an aqueous core to allow the encapsulation and protection of hydrophilic molecules such as miRNA or DNA (62).…”

The nanostructured secretome

“…This may imply the use of various synthetic components and synthesis techniques, which range from the exogenous loading of drug molecules through electroporation to the production of extracellular vesicle by direct cell extrusion. In secNP modification, very often tweaking and engineering overlap and/or combine, 101 as we will concisely present in the next subsections.…”