Self-programmed enzyme phase separation and multiphase coacervate droplet organization

Karoui, Hedi; Seck, Marianne J.; Martin, Nicolas

doi:10.1039/d0sc06418a

Cited by 51 publications

(42 citation statements)

References 51 publications

(69 reference statements)

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“…The time-programmed phase behavior is currently available by changes in pH, temperature, ionic strength, light (UV), and more recently by enzyme-mediated catalytic activity ( Shin et al, 2017 ; Schuster et al, 2018 ; Martin et al, 2019 ; Wheeler et al, 2019 ; Garabedian et al, 2021 ). Furthermore, by converting chemical fuels, the coacervate droplet could behave like a protocell capable of self-division, making it an ideal model for approaching the dynamic complexity of living cells ( Zwicker et al, 2017 ; Donau et al, 2020 ; Karoui et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Liquid-Liquid Phase Separation: Unraveling the Enigma of Biomolecular Condensates in Microbial Cells

et al. 2021

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Numerous examples of microbial phase-separated biomolecular condensates have now been identified following advances in fluorescence imaging and single molecule microscopy technologies. The structure, function, and potential applications of these microbial condensates are currently receiving a great deal of attention. By neatly compartmentalizing proteins and their interactors in membrane-less organizations while maintaining free communication between these macromolecules and the external environment, microbial cells are able to achieve enhanced metabolic efficiency. Typically, these condensates also possess the ability to rapidly adapt to internal and external changes. The biological functions of several phase-separated condensates in small bacterial cells show evolutionary convergence with the biological functions of their eukaryotic paralogs. Artificial microbial membrane-less organelles are being constructed with application prospects in biocatalysis, biosynthesis, and biomedicine. In this review, we provide an overview of currently known biomolecular condensates driven by liquid-liquid phase separation (LLPS) in microbial cells, and we elaborate on their biogenesis mechanisms and biological functions. Additionally, we highlight the major challenges and future research prospects in studying microbial LLPS.

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Section: Discussionmentioning

confidence: 99%

Liquid-Liquid Phase Separation: Unraveling the Enigma of Biomolecular Condensates in Microbial Cells

et al. 2021

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“…Essential elements of membraneless organelles can be reconstituted in vitro, typically based on disordered proteins with weak-multivalent interactions 21 . Reconstituted and engineered condensates have been shown to be able to partition enzymes 14,[22][23][24][25][26][27][28] , and therefore their activity.…”

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confidence: 99%

Sustained enzymatic activity and flow in crowded protein droplets

et al. 2021

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Living cells harvest energy from their environments to drive the chemical processes that enable life. We introduce a minimal system that operates at similar protein concentrations, metabolic densities, and length scales as living cells. This approach takes advantage of the tendency of phase-separated protein droplets to strongly partition enzymes, while presenting minimal barriers to transport of small molecules across their interface. By dispersing these microreactors in a reservoir of substrate-loaded buffer, we achieve steady states at metabolic densities that match those of the hungriest microorganisms. We further demonstrate the formation of steady pH gradients, capable of driving microscopic flows. Our approach enables the investigation of the function of diverse enzymes in environments that mimic cytoplasm, and provides a flexible platform for studying the collective behavior of matter driven far from equilibrium.

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“…Albeit still largely empirical, sequestration by coacervates is usually highly selective so that solutes can either be accumulated within droplets or excluded from them. [108] Coacervate droplets can also be dynamically formed and dissolved in response to a range of cues, [109] including light [110,111] and enzyme reactions, [112] offering an additional level of control over solute compartmentalization. Significantly, the local up-concentration of reactive species has been exploited to demonstrate enhanced enzyme-based reactions, [113] ribozyme catalysis, [49,50] or template-directed RNA polymerization.…”

Section: Coacervate Microdroplets As Protocell Modelsmentioning

confidence: 99%

Fatty Acid Vesicles and Coacervates as Model Prebiotic Protocells

Martin

Douliez

2021

ChemSystemsChem

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The prebiotic organization of chemicals into compartmentalized ensembles is an essential step to understand the transition from inert molecules to living matter. Compartmentalization is indeed a central property of living systems. Fatty acids represent the simplest prebiotic amphiphiles capable of selfassembling into membrane-bound vesicles, and have therefore emerged as valuable molecules to create models of protocellular compartments. Here, the main experimental findings supporting this idea are reviewed, together with approaches to increase the stability of fatty acid vesicles in adverse pH, salt, or temperature conditions. Recent studies on the self-assembly of fatty acids into membrane-free coacervate microdroplets are then discussed, providing a promising new paradigm for prebiotic compartmentalization. Lastly, it is also argued that the unique possibility of cycling between fatty acid vesicles and coacervates paves the way to an exciting new hypothesis for the emergence of the first living protocells.

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Self-programmed enzyme phase separation and multiphase coacervate droplet organization

Abstract: Self-programmed enzyme phase separation is exploited to assemble dynamic multiphase coacervate droplets via spontaneous polyion self-sorting under non-equilibrium conditions.

Cited by 51 publications

References 51 publications

Liquid-Liquid Phase Separation: Unraveling the Enigma of Biomolecular Condensates in Microbial Cells

Liquid-Liquid Phase Separation: Unraveling the Enigma of Biomolecular Condensates in Microbial Cells

Sustained enzymatic activity and flow in crowded protein droplets

Fatty Acid Vesicles and Coacervates as Model Prebiotic Protocells

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