PLANT-DERIVED VESICLES INCORPORATING TRANS-MEMBRANE PROTEINS
The disclosure relates to a vesicle in a liquid composition, the vesicle including plant-derived transmembrane proteins. The disclosed embodiments further relate to methods of producing the vesicles and separation membranes of such vesicles. The present disclosure provides for the preparation of a separation membrane using vesicle components originating exclusively or mainly from natural sources.
1 . A vesicle in a liquid composition, said vesicle comprising plant-derived transmembrane proteins, wherein the vesicle-forming material comprises plant plasma membrane components.
2 . The vesicle according to claim 1 , wherein the vesicle forming material consists of plant plasma membrane components.
3 . The vesicle according to claim 1 , wherein the plant plasma membrane components originate from the same plant as the plant-derived transmembrane proteins.
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7 . The vesicle according to claim 1 , wherein the liquid composition comprises a reactive end group functionalised PDMS (polydimethylsilane).
8 . The vesicle according to claim 1 , wherein said reactive end group functionalised PDMS is functionalized with one or more of amine, carboxylic acid, and/or hydroxy group(s), and wherein the number of monomers is in the range of 30 to 50.
9 . The vesicle according to claim 1 , wherein the reactive end group functionalised PDMS is poly(dimethylsiloxane), bis(3-aminopropyl).
10 . The vesicle according to claim 1 , wherein the plant-derived components comprise plant plasma membrane phospholipids.
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15 . The vesicle according to claim 1 , wherein the liquid composition does not comprise a detergent or a surfactant.
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17 . The vesicle according to claim 1 for use in a separation membrane.
18 . A method of preparing the vesicles in a liquid composition according to claim 1 , comprising extracting the plant plasma membrane components and the plant-derived transmembrane proteins from a plant using ultracentrifugation and/or two-phase partitioning.
19 . The method according to claim 18 , comprising mixing the extracted plant-derived transmembrane proteins with a basic formulation obtained by dissolving polyethylene glycol-hydroxystearate in phosphate-buffered saline solution.
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22 . The method according to claim 18 , wherein the total protein concentration of the liquid composition is below 10 mg/ml.
23 . The method according to claim 22 , wherein the total water channel transmembrane protein concentration of the liquid composition is between 0.01 μg/ml and 100 μg/ml.
24 . A separation membrane comprising the vesicle according to claim 1 .
25 . The separation membrane according to claim 24 , wherein the separation membrane comprises an active layer incorporating the vesicle and a porous support membrane.
26 . The separation membrane according to claim 25 , wherein the active layer comprises the vesicle incorporated in a thin film composite (TFC) layer formed on a porous substrate membrane.
27 . The separation membrane according to claim 26 , wherein the TFC layer is formed by interfacial polymerisation between a di-amine or tri-amine monomer compound and an acyl halide monomer compound.
28 . The separation membrane according to claim 27 , wherein the vesicles are immobilized and/or chemically bound to the TFC layer by naturally occurring free reactive groups, such as amino groups, hydroxyl groups and carboxyl groups, on the surface of the vesicles.
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31 . A method of preparing a thin film composite layer immobilizing vesicles incorporating a transmembrane protein on a porous substrate membrane, comprising
a. providing a mixture of vesicles in a liquid composition according to any one of the claims 1 to 17 and a di-amine or tri-amine compound,
b. covering the surface of a porous support membrane with the mixture of a,
c. applying a hydrophobic solution comprising an acyl halide compound, and
d. allowing the aqueous solution and the hydrophobic solution to perform an interfacial polymerization reaction to form the thin film composite layer.
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33 . The method according to claim 31 , wherein the proportion of the di-amine or tri-amine compound to acyl halide compound is from 0:1 to 30:1 by weight.
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