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

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. Here we present a microbial platform for high yield production of lipidic nanovesicles, with clickable thiol moieties in their outer corona. These nanovesicles show low size dispersity, are decorated with a dense, perfectly oriented and customizable corona of transmembrane polypeptides. In addition, 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 taking advantage of the diversity of commercially-available maleimide derivatives, engineering bacterial-based proteoliposomes are an attractive eco-friendly alternative that can outperform current liposome preparation methods.