Process of obtaining thylakoids from photosynthetic organisms; plant fractions obtained from the process; pure thylakoids; and methods of use of thylakoids as ROS scavengers, photo-protectors, biosensors, biofilters and bioreactors
View Patent ↗This invention relates to a process by which an extract comprising integral thylakoids is obtained. The resulting extract is a potent dynamic antioxidant useful as a ROS (reactive oxygen species) scavenger. This extract is intended to be used for the treatment or prevention of diseases involving the generation of ROS, such as inflammatory diseases or cancer. This extract also finds a use as a solar screen because of its capacity to capture UV radiations and to dissipate the solar energy into heat.
1. An extract from a photosynthetic organism obtainable by a method comprising:
(a) providing a suspension of photosynthetic organism constituents that contain thylakoids;
(b) disrupting the constituents while recovering thylakoids under light conditions which minimize light flux, in a medium comprising sugar or sugar alcohol and having a pH above 2 and below 10, the medium being added in a volume calculated upon the following equation:
(Volume of medium+organism constituents water content)/(Organism constituents dry weight)>10
to obtain a composition consisting essentially of thylakoids, cell debris/membranes and a liquid phase;
(c) separating and isolating an extract from the cell debris/membranes and the liquid phase, wherein the separated and isolated extract consists essentially of pure stabilized integral thylakoids, the thylakoids comprising organized photosynthetic pigments in their fundamental state; and
(d) eliminating any electron donor from the extract.
2. The extract according to claim 1 , wherein the equation is:
(Volume of medium+organism constituents water content)/(Organism constituents dry weight)=25−150.
3. The extract according to claim 1 , wherein the pH is between 5 and 8.
4. The extract according to claim 1 , wherein the pH is between 6 and 7.5.
5. The extract of claim 1 , wherein said organism is a plant.
6. The extract according to claim 5 , wherein the suspension of step (a) is obtained by mechanically dispersing plant constituents or tissues in said medium.
7. The extract according to claim 5 , wherein step (a) is preceded by a step of submitting a plant to a conditioning parameter selected from light, osmotic stress, heat, cold, freezing, dryness, hormones, chemical and biological inducers.
8. The extract according to claim 1 , wherein step (a) is preceded by a step of conditioning said organism in a light environment of a wavelength comprised between about 500 and 600 nm, and step (b) is performed under the same light conditions.
9. The extract according to claim 1 , wherein the medium comprises sorbitol in a concentration of about 0.2 to 1.5 M.
10. The extract according to claim 1 , wherein the medium comprises sorbitol in a concentration of about 0.2 to 0.4 M.
11. The extract according to claim 1 , wherein said medium comprises a buffer selected from a Tris buffer, an acetate buffer and an ascorbate buffer.
12. The extract according to claim 1 , wherein said medium comprises about 20 mM Tris or acetate or ascorbate buffer having a pH of 7.5 and about 350 mM sorbitol or sucrose or fructose.
13. The extract according to claim 1 , wherein the separation is based on a difference of sedimentation coefficient of each of the thylakoids, the cell debris/membranes, and the liquid phase.
14. The extract according to claim 13 , wherein the separation comprises centrifugation in a tube equipped with a filter in a superior portion of the tube, the filter having a porosity onto which the cell debris/membranes deposit while the thylakoids and the liquid phase pass through the filter, the thylakoids forming a pellet in an inferior portion of the tube.
15. The extract according to claim 1 , wherein water is eliminated by exchanging it against an amphoteric solvent or surfactant after step (c).
16. The extract according to claim 15 , wherein water is eliminated by exchanging it against propylene glycol.
17. The extract according to claim 1 that is in a state selected from a dry state, a partly or totally solubilized state, and a suspension.
18. A composition comprising the extract according to claim 1 .