Growing Membranes <i>In Vitro</i> by Continuous Phospholipid Biosynthesis from Free Fatty Acids

Exterkate, Marten; Caforio, Antonella; Stuart, Marc C. A.; Driessen, Arnold J. M.

doi:10.1021/acssynbio.7b00265

Cited by 70 publications

(96 citation statements)

References 39 publications

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“…[98] In the latter study, the enzymes and precursors were not encapsulated and synthesis occurred outside the liposomes. [98] In the latter study, the enzymes and precursors were not encapsulated and synthesis occurred outside the liposomes.…”

Section: Growth Via Uptake Of Synthesized Membrane Componentsmentioning

confidence: 99%

Directed Growth of Biomimetic Microcompartments

et al. 2019

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twilight zone), it is widely recognized that the formation of micro-and nanocompartments is an essential ingredient of all forms of life. This spatial constraint serves numerous purposes, including the segregation and protection from the environment (to allow for individuality and maintenance), the establishment of gradients (to enable and make use of outof-equilibrium conditions), and the role of 2D and 3D confinement for self-organization phenomena, all of which serve to overcome the overall "dilution problem." In order to fulfill another cornerstone of life-proliferation-compartments also need to change with time by fusion, growth, and division. In particular, the dynamic growth of compartments is an essential prerequisite for enabling selfreproduction as a fundamental life process, both in simplistic systems such as droplets or fatty acid-based vesicles, as well as for lipid vesicle compartments with membranes that resemble the biomembranes of today's cells. In this context, we argue that growth deserves more attention, not only because growth precedes division but also because of the difficulty to realize growth compared to division, especially in the case of lipid vesicles, where budding and division has been observed in response to various factors. This growth aspect has also been recognized in basic theoretical models of living systems such as Ganti's chemoton, [1,2] for which the increase of membrane area (referred to as membrane formation) was postulated to be one of three subsystems, characterizing living entities from a chemical viewpoint. The autopoietic theory was also centered on the membrane but focused on self-maintenance rather than on (self-)reproduction. [3,4] However, such a self-maintaining system could also enter a self-reproduction mode, manifested by growth, if homeostatic misbalance leads to excess membrane formation as shown in a thought experiment by Luisi. [3] In this progress report, we review the existing approaches for growth of compartments in the context of bottom-up synthetic biology and protobiology. We consider mainly micrometer-sized compartments due to their characteristic cellular dimensions; this feature also ensures that the area and curvature of the interface or the bounding membrane has less influence on the enclosed solution. Microcompartments of various origins and chemistries have been used as protocell models, and many studies have addressed growth and division simultaneously in an ambitious effort to mimic self-reproduction. Contemporary biological cells are sophisticated and highly compartmentalized. Compartmentalization is an essential principle of prebiotic life as well as a key feature in bottom-up synthetic biology research.In this review, the dynamic growth of compartments as an essential prerequisite for enabling self-reproduction as a fundamental life process is discussed. The micrometer-sized compartments are focused on due to their cellular dimensions. Two types of compartments are considered, membraneless droplets and membrane-bound microcompartments. Gr...

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Section: Growth Via Uptake Of Synthesized Membrane Componentsmentioning

confidence: 99%

Directed Growth of Biomimetic Microcompartments

et al. 2019

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“…[74] Startingf rom oleic acid and glycerol, the intermediate CDP-DAGi sf ormed in four enzymatic steps, after which further conversion yields either DOPE or DOPG ( Figure 6). This lipid composition supports high rates of transport of the bacterialt ransporters that we have studied;f or eukaryotic membrane proteins as terol and some specific lipids might be required (see Section 4.1);w ed on ot consider this here.…”

Section: Lipid Synthesis For Compartmentalizationmentioning

confidence: 99%

Cell Fuelling and Metabolic Energy Conservation in Synthetic Cells

et al. 2019

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We are aiming for a blue print for synthesizing (moderately complex) subcellular systems from molecular components and ultimately for constructing life. However, without comprehensive instructions and design principles, we rely on simple reaction routes to operate the essential functions of life. The first forms of synthetic life will not make every building block for polymers de novo according to complex pathways, rather they will be fed with amino acids, fatty acids and nucleotides. Controlled energy supply is crucial for any synthetic cell, no matter how complex. Herein, we describe the simplest pathways for the efficient generation of ATP and electrochemical ion gradients. We have estimated the demand for ATP by polymer synthesis and maintenance processes in small cell‐like systems, and we describe circuits to control the need for ATP. We also present fluorescence‐based sensors for pH, ionic strength, excluded volume, ATP/ADP, and viscosity, which allow the major physicochemical conditions inside cells to be monitored and tuned.

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“…There also have been various attempts to reconstitute parts of the natural phospholipid synthesis pathway in the context of artificial cell membranes . Phospholipid synthesis involves membrane‐inserted enzymes such as sn‐glycerol‐3‐phosphate acyltransferase (GPAT, PlsB in E.coli ) and lysophosphatidic acid acyltransferase (LPAAT, PlsC in E.coli ), which convert glycerol‐3‐phosphate (G3P) into phosphatidic acid (PA), which can be further transformed into phosphocholine (PC) and phosphoethanolamine (PE) lipids.…”

Section: Growth and Division Of Cell‐sized Compartmentsmentioning

confidence: 99%

“…Phospholipid synthesis involves membrane‐inserted enzymes such as sn‐glycerol‐3‐phosphate acyltransferase (GPAT, PlsB in E.coli ) and lysophosphatidic acid acyltransferase (LPAAT, PlsC in E.coli ), which convert glycerol‐3‐phosphate (G3P) into phosphatidic acid (PA), which can be further transformed into phosphocholine (PC) and phosphoethanolamine (PE) lipids. Using PlsB, PlsC and the water‐soluble enzyme FadD10, which catalyzes the adenylation of long‐chain fatty acids into fatty acid adenylates, membrane expansion of liposomes has been demonstrated via cell‐free phospholipid production starting from oleic acid . Recently, a minimal biochemical route towards the creation of phospholipids was demonstrated also involving FadD10, in which FA adenylates were shown to spontaneously react with amine‐functionalized lysolipids to form phospholipids .…”

Section: Growth and Division Of Cell‐sized Compartmentsmentioning

confidence: 99%

Genetically Encoded Membranes for Bottom‐Up Biology

2019

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The creation of self‐replicating cell‐mimicking systems – artificial cells – is one of the major goals of bottom‐up synthetic biology. An essential aspect of such systems is the realization of membranous compartments which can grow and divide in synchrony with the internal dynamics of the cells. Amphiphiles capable of forming membranes may be either externally provided to feed the growing compartments, or generated in situ through chemical processes. In the context of autonomously self‐replicating systems, genetically encoded membranes are of particular interest. Herein, we discuss typical approaches taken for the creation of cell‐like microcompartments via self‐assembly of amphiphiles. We specifically address some of the challenges associated with the generation of phospholipid or peptide‐based membranes via genetic and enzymatic processes.

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Growing Membranes In Vitro by Continuous Phospholipid Biosynthesis from Free Fatty Acids

Cited by 70 publications

References 39 publications

Directed Growth of Biomimetic Microcompartments

Directed Growth of Biomimetic Microcompartments

Cell Fuelling and Metabolic Energy Conservation in Synthetic Cells

Genetically Encoded Membranes for Bottom‐Up Biology

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