Isolation of chromoplastid carotenoids from fruits
AssigneeUniversita Degli Studi di Cagliari
InventorsDario Piano, Domenica Farci
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Abstract
The present invention relates to a method for the production of chromoplastid carotenoids that can be isolated from fruits. The procedure of the invention allows to selectively isolate, on the basis of the size (Size Exclusion Chromatography) and in pure form, the carotenoid in the form of regularly shaped and sized aggregates. The process includes the following steps: i) production, in suspension, of homogeneous fragments of the chromoplastidial membranes from the fruits; ii) solution isolation of the membrane components by solubilization with detergents; iii) selective and high-purity isolation of the carotenoid in the form of regular paracrystalline aggregates by a screening technique such as size exclusion chromatography (SEC—Size Exclusion Chromatography).
Claims15
A process for obtaining a chromoplastid carotenoid from a plant fruit comprising the following steps:
- i) preventive washing and crushing of the plant fruit;
- (ii) fragmenting mechanically the washed and crushed fruit in a first buffered solution;
- (iii) centrifuging to generate a water-soluble cytosolic fraction and a solid fraction to separate out the water-soluble cytosolic fraction;
- (iv) recovering the separated solid fraction comprising a chromoplastidial membrane after the centrifugation, and dispersion of the separated solid fraction in a second buffered solution,
- (v) adding to the second buffered solution having dispersed therein the separate solid fraction an alkylsaccharide detergent, thereby solubilizing the chromoplastid carotenoid, and mixing and subsequent centrifuging to generate a supernatant solution;
- and
- (vi) removing the supernatant solution after the centrifugation of step (v), and
- (vii) isolating the chromoplastid carotenoid from the removed supernatant solution by a process selected from the group consisting of:
- Size Exclusion Chromatography, Fast Protein Liquid Chromatography, ultracentrifugation, nano- and ultrafiltration.
A process for separating a chromoplastid carotenoid from a plant material comprising the following steps:
- i) preventive washing and crushing of the plant material;
- (ii) fragmenting mechanically the washed and crushed plant material in a first buffered solution;
- (iii) centrifuging to generate a water-soluble cytosolic fraction and a solid fraction to separate out the water-soluble cytosolic fraction;
- (iv) recovering the separated solid fraction comprising a chromoplastidial membrane after the centrifugation, and dispersion of the separated solid fraction in a second buffered solution,
- (v) adding to the second buffered solution having dispersed therein the separate solid fraction an alkylsaccharide detergent, and mixing the resultant solution, thereby solubilizing the chromoplastid carotenoid,
- (vi) centrifuging the solution generated after missing in step (v) to generate a supernatant solution;
- and
- (vi) removing the supernatant solution after the centrifugation step (vi).
Dependencies are inferred from the claim text.