Abstract
Light‐driven ATP regeneration systems combining ATP synthase and bacteriorhodopsin have been proposed as an energy supply in the field of synthetic biology. Energy is required to power biochemical reactions within artificially created reaction compartments like protocells, which are typically based on either lipid or polymer membranes. The insertion of membrane proteins into different hybrid membranes is delicate, and studies comparing these systems with liposomes are needed. Here we present a detailed study of membrane protein functionality in different hybrid compartments made of graft polymer PDMS‐g‐PEO and diblock copolymer PBd‐PEO. Activity of more than 90 % in lipid/polymer‐based hybrid vesicles could prove an excellent biocompatibility. A significant enhancement of long‐term stability (80 % remaining activity after 42 days) could be demonstrated in polymer/polymer‐based hybrids.
Highlights
Introduction by the proton-gradient driven enzyme FOF1-ATP synthase
An artificial light-driven ATP regeneration module has been built through bottom-up assembly of purified transmembrane proteins
Bacteriorhodopsin, a light-driven proton pump, establishes a proton gradient to drive the synthesis of ATP by FOF1ATP synthase from Escherichia coli. The reconstitution of both enzymes into phosphatidylcholine (PC) unilamellar vesicles is optimized for performance using a Triton X-100 mediated reconstitution into preformed liposomes, similar to the method described by Fischer et al.[11d]. The ATP production rate in the natural phospholipid environment is later used as a benchmark for the evaluation of biocompatibility of both transmembrane proteins in different hybrid vesicles
Summary
Introduction by the proton-gradient driven enzyme FOF1-ATP synthase. For mimicking natural photophosphorylation, a series of artificial systems have been constructed to capture the energy of light and move protons across the membrane.[9]. In the framework of bottom-up synthetic biology, new exciting possibilities for the combination of natural and synthetic based materials arise. In this regard, Choi and Montemagno[12] reported the first successful incorporation of bacteriorhodopsin (bR) and ATP synthase from thermophilic. The proposed approach employs mixed hybrid vesicles from both copolymers and lipids, enabling fine-tuning of the membrane physical properties.[14] A recent example demonstrates the functional incorporation of cytochrome bo quinol oxidase (bo oxidase) in such hybrid vesicles, consisting of diblock copolymer polybutadiene-b-poly(ethylene oxide)
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