Constructing artificial respiratory chain in polymer compartments: Insights into the interplay betweenbo3oxidase and the membrane

Marušič, Nika; Otrin, Lado; Zhao, Ziliang; Lira, Rafael B.; Kyrilis, Fotis L.; Hamdi, Farzad; Kastritis, Panagiotis L.; Vidaković‐Koch, Tanja; Ivanov, Ivan; Sundmacher, Kai; Dimova, Rumiana

doi:10.1073/pnas.1919306117

Cited by 50 publications

(118 citation statements)

References 87 publications

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“…In 2018, we achieved increased stability of cytochrome bo 3 reconstituted in hybrid vesicles up to 500 days [197]. Similarly, recent work from Dimova group showed that functional integration of cytochrome bo 3 oxidase in synthetic membranes made of PDMS-g-PEO was capable of lumen acidification and such reconstituted system showed to increase the active lifetime and resistance to free radicals [198]. In 2017, Otrin et al [199] demonstrated a similar ability to store gradients by reconstituting cytochrome bo 3 together with an ATP synthase in hybrid vesicles constituted of the same copolymer, PDMS-g-PEO.…”

Section: Extending the Lifetime Of Membrane Enzymesmentioning

confidence: 63%

Membrane Protein Modified Electrodes in Bioelectrocatalysis

2020

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Transmembrane proteins involved in metabolic redox reactions and photosynthesis catalyse a plethora of key energy-conversion processes and are thus of great interest for bioelectrocatalysis-based applications. The development of membrane protein modified electrodes has made it possible to efficiently exchange electrons between proteins and electrodes, allowing mechanistic studies and potentially applications in biofuels generation and energy conversion. Here, we summarise the most common electrode modification and their characterisation techniques for membrane proteins involved in biofuels conversion and semi-artificial photosynthesis. We discuss the challenges of applications of membrane protein modified electrodes for bioelectrocatalysis and comment on emerging methods and future directions, including recent advances in membrane protein reconstitution strategies and the development of microbial electrosynthesis and whole-cell semi-artificial photosynthesis.

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Section: Extending the Lifetime Of Membrane Enzymesmentioning

confidence: 63%

Membrane Protein Modified Electrodes in Bioelectrocatalysis

2020

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“…Even after three complete light-dark cycles, we observed only little decrease in the pH gradient. Compared to previous reports where proton pumps were reconstituted in lipid vesicles, [7,27] we could achieve faster and higher pH gradients using genetically engineered E. coli. Moreover, the use of E. coli circumvented the need for cumbersome protein purification and reconstitution to prepare proteoliposomes, [28] which highlights a key advantage of merging top-down and bottom-up synthetic biology.…”

Section: Top-down Engineering Of E Coli For Light-harvestingmentioning

confidence: 87%

Actuation of synthetic cells with proton gradients generated by light-harvesting E. coli

Jahnke

Ritzmann

Fichtler

et al. 2020

Preprint

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Bottom-up and top-down approaches to synthetic biology each employ distinct methodologies with the common aim to harness new types of living systems. Both approaches, however, face their own challenges towards biotechnological and biomedical applications. Here, we realize a strategic merger to convert light into proton gradients for the actuation of synthetic cellular systems. We genetically engineer E. coli to overexpress the light-driven inward-directed proton pump xenorhodopsin and encapsulate them as organelle mimics in artificial cell-sized compartments. Exposing the compartments to light-dark cycles, we can reversibly switch the pH by almost one pH unit and employ these pH gradients to trigger the attachment of DNA structures to the compartment periphery. For this purpose, a DNA triplex motif serves as a nanomechanical switch responding to the pH-trigger of the E. coli. By attaching a polymerized DNA origami plate to the DNA triplex motif, we obtain a cytoskeleton mimic that considerably deforms lipid vesicles in a pH-responsive manner. We foresee that the combination of bottom-up and top down approaches is an efficient way to engineer synthetic cells as potent microreactors.

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“…[ 130–135 ] A comprehensive comparison between lipid and polymer vesicles from physical properties to cellular function reconstructions can be found in refs. [ 136,137 ] .…”

Section: Compartmentalization Via Interfaces Of Multiphase Systemsmentioning

confidence: 99%

“…[ 161 ] Surprisingly, hybrid vesicles with intermediate lipid/copolymer ratios can be more permeable to ion transport than pure lipid and polymer vesicles, arising from dynamic equilibrium of vesicles and tubular micelle formations. [ 137,163 ] Hybrid vesicles also allow the accommodation of hydrophobic proteins and nanoparticles more easily than pure lipid vesicles, [ 44,167 ] and hydrophilic nanoparticles cannot be accommodated since they appears either in the interior or the outlier of the bilayer of the vesicle. The size and surface charge of the hydrophobic nanoparticle affect the topology of the bilayer vesicle differently.…”

Section: Compartmentalization Via Interfaces Of Multiphase Systemsmentioning

confidence: 99%

“…[ 135 ] A recent discovery showed that the proton permeability of hybrid membrane is higher than that of liposomes and polymersomes, and the proton permeability of microscopically heterogenous phase‐separated hybrid membranes is higher than that of homogeneous ones. [ 137 ] Unexpectedly, the insertion of bo 3 oxidase, chemically driven proton pump, into liquid–copolymer hybrid membrane can cause the membrane rearrangement and resealing, which decreases the proton permeability of the hybrid membrane. [ 137 ]…”

Section: Cell‐like Biological Functions At Different Levelsmentioning

confidence: 99%

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Interface Engineering in Multiphase Systems toward Synthetic Cells and Organelles: From Soft Matter Fundamentals to Biomedical Applications

et al. 2020

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products of evolution over billions of years. They are featured by multiple hierarchical compartments performing specialized and exquisitely coordinated functions. The biological and genetic complexity of cells and subcellular components make understanding their physicochemical foundation piece by piece challenging. [1-5] Moreover, the mystery of how the very first cell, protocell, comes into exist in early earth scenario is unveiled yet. [6] Motivated by understanding protocell and biological cells, synthetic cells are being constructed. Started form the simplest form, aqueous droplet enclosed by a bilayer structure, researchers adopt a bottom-up approach to study machineries of biological cells, and explore possible forms of the protocell in early world conditions. [7] Synthetic cells are important to bridge the gap between cellular organism and nonliving matter. They refer to synthetic compartments that encapsulate a wide variety of molecules and mimic one or multiple cellular functions. By segregation of contents in different compartments, synthetic cells are now engineered to perform multiple processes and functions, including segregating genetic information, genetic circuits, protein synthesis, metabolism, growth, reproduce, recognition, intercellular crosstalk, and adaptivity to surroundings. [8-17] In these simplified cell models, cellular processes and signal pathways are reconstituted, providing insights for cell biology and offering new opportunities for designing smart cell-like carriers of therapeutics. [18-26] In addition, the hypothesis of "RNA world" has inspired the investigation of synthetic compartments as protocells, in which nucleotide permeation, compartmental division, the synthesis of macromolecular polynucleotide, and the polymerization of ribonucleic acids (RNA) are explored. [7,27,28] The beginning of synthesizing cell-like structures starts from amphiphiles self-assembled at liquid/liquid interfaces to form enclosed compartments. [29-32] Recently, techniques such as microfluidics facilitate the fabrication of complex compartments with high-order hierarchy with excellent control. [33-36] Formulations of compartmentalization can be diverse, there are reviews mainly focused on one particular type of synthetic compartments, such as lipid vesicles (liposomes), [22,30,37,38] polymer vesicles (polymersomes), [39-43] lipid-polymer hybrid Synthetic cells have a major role in gaining insight into the complex biological processes of living cells; they also give rise to a range of emerging applications from gene delivery to enzymatic nanoreactors. Living cells rely on compartmentalization to orchestrate reaction networks for specialized and coordinated functions. Principally, the compartmentalization has been an essential engineering theme in constructing cell-mimicking systems. Here, efforts to engineer liquid-liquid interfaces of multiphase systems into membrane-bounded and membraneless compartments, which include lipid vesicles, polymer vesicles, colloidosomes, hybrids, and coacervate droplets,...

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Constructing artificial respiratory chain in polymer compartments: Insights into the interplay betweenbo₃oxidase and the membrane

Cited by 50 publications

References 87 publications

Membrane Protein Modified Electrodes in Bioelectrocatalysis

Membrane Protein Modified Electrodes in Bioelectrocatalysis

Actuation of synthetic cells with proton gradients generated by light-harvesting E. coli

Interface Engineering in Multiphase Systems toward Synthetic Cells and Organelles: From Soft Matter Fundamentals to Biomedical Applications

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