In vitro gene expression and detergent-free reconstitution of active proteorhodopsin in lipid vesicles

Abstract

In situ generation of proton gradients across membranes is a key component for energy generation within cells and is therefore an important feature for the design of energy converting artificial cells. Here, we establish a stepwise method for the in vitro expression and detergent-free reconstitution of proteorhodopsin into the membrane of lipid vesicles. This represents a novel technique towards the bottom-up synthesis of energy-generating artificial cells. Impact statement Our results offer the potential for straightforward, additive-free, and molecularly simple routes to building complex bioreactors based on in vitro transcription–translation systems and lipid vesicles.