Methods for isolating compounds
The present invention in its broadest aspect relates to a method for reducing glycoalkaloid content and turbidity of an aqueous phase comprising compounds selected from two or more of PA, PI, PPO, LipO, pectin, lipid, glycoalkaloid and phenolic compounds of which at least one compound is selected from PA, PT, LipO and PPO; a) providing an aqueous phase comprising compounds selected from two or more of PA, PI, PPO, LipO, pectin, lipid, glycoalkaloid and phenolic compounds of which at least one compound is selected from PA, PT, LipO and PPO; and b) performing one or more steps to reduce the concentration of solanine in the dry matter of the aqueous phase with at least 15 percent, such as at least 20%, such as at least 25% and to achieve an optical density at 620 nm of the remaining aqueous phase of less than 0.7; such as less than 0.5; such as less than 0.3; such as less than 0.2; such as less than 0.1; and thereby obtaining an aqueous phase having reduced glycoalkaloid content and turbidity compared to an untreated aqueous phase.
1. A method for isolating a first group of compounds selected from one or more of patatin protein (PA), protease inhibitor protein (PI), lipoxygenase (LipO) and polyphenol oxidase (PPO) from a second group of compounds selected from one or more of PA, PI, LipO, PPO, glycoalkaloid and phenolic compounds, said method comprising:
a) providing an aqueous phase comprising compounds selected from two or more of PA, PI, PPO, LipO, glycoalkaloid and phenolic compounds of which at least one compound is selected from one or more of PA, PI, LipO and PPO;
b) contacting the aqueous phase with a mobile solubilized ligand at physico-chemical conditions allowing formation of a complex between the ligand and the compounds selected from one or more of PA, PI, LipO and PPO;
c) allowing the complex to separate from the aqueous supernatant phase, optionally by changing said physico-chemical conditions in the composition to reduce the stability of the complex; and
d) isolating the complex separated from the aqueous supernatant phase by precipitation.
2. The method of claim 1 , wherein the separated complex comprises a combination of PA and PI and PPO and wherein the complex separates from the aqueous supernatant phase by precipitation.
3. The method of claim 2 , wherein the dry weight ratio PA:PI in the precipitate is higher than the dry weight PA:PI ratio for PA and PI remaining dissolved in the aqueous supernatant phase.
4. The method of claim 3 , further comprising dissolving the precipitated complex of step d) in an aqueous solvent and isolating PA from one or more compounds selected from PI and PPO by a mechanical separation process concentrating the PA in the retentate.
5. The method of claim 3 , further comprising isolating the precipitated complex by a mechanical separation process concentrating one or more of PA, PI and PPO in the retentate.
6. The method of claim 3 , further comprising:
a) contacting the aqueous supernatant phase with a further mobile solubilized ligand at physico-chemical conditions in the aqueous supernatant phase allowing formation of a complex between the ligand and compounds selected from one or more of PI, and PPO;
b) allowing the complex to separate from the aqueous supernatant phase, optionally by changing the physico-chemical conditions in the composition to reduce the solubility of the complex so that the complex separates from the aqueous supernatant phase; and
c) isolating the complex.
7. The method of claim 2 , wherein the sum of PA and PI remaining dissolved in the aqueous supernatant phase is less than 20%, optionally less than 10 wt. % of the total PA and PI.
8. The method of claim 7 , further comprising dissolving the precipitated complex in an aqueous solvent and isolating PA from one or more compounds selected from PI, and PPO by a mechanical separation process concentrating the PA in the retentate.
9. The method of claim 7 , further comprising isolating the precipitated complex by mechanical separation process concentrating the one or more or PA, PI, and PPO in the retentate.
10. The method of claim 2 , wherein the dry weight ratios PI:PA or PPO:PA in the precipitate is higher than the dry weight ratios PI:PA or PPO:PA for PA, Pi and PPO remaining dissolved in the aqueous supernatant phase.
11. The method of claim 10 , further comprising concentrating PA in the aqueous supernatant phase by mechanical separation process concentrating PA in the retentate, optionally combined with diafiltration.
12. The method of claim 10 further comprising:
a) contacting the aqueous supernatant phase with a further mobile solubilized ligand at physico-chemical conditions in aqueous supernatant phase allowing formation of a complex between the ligand and PA;
b) allowing the complex to separate from the aqueous supernatant phase, optionally by changing said physico-chemical conditions in the composition to reduce the solubility of the complex so that the complex separates from the aqueous supernatant phase; and
c) isolating the complex.
13. The method of claim 10 , further comprising adsorbing dissolved PA in the aqueous supernatant phase on an immobilized solid carrier at conditions where the carrier will bind PA.
14. The method of claim 1 , further comprising pre-treating the aqueous phase of a) by absorbing one or more of PI, LipO and PPO on an immobilized solid carrier at conditions where the carrier will bind the one or more of PI, LipO or PPO.
15. The method of claim 1 , wherein the PA is complexed to the ligand in a PA: ligand dry weight ratio of at least 4:1.
16. The method of claim 1 , wherein the PI is complexed to the ligand in a PI: ligand dry weight ratio of at least 3:1.
17. The method of claim 1 , wherein the PPO is complexed to the ligand in a PPO: ligand dry weight ratio of at least 2:1.
18. The method of claim 1 , wherein the aqueous phase of step a) comprise an aqueous solution liberated when disintegrating a portion of a plant optionally a tuber portion of a plant of the genus Solanum , optionally of the species S. tuberosum.
19. A method for isolating one or more of glycoalkaloid, LipO and phenolic compounds said method comprising:
a) providing an aqueous phase comprising one or more of glycoalkaloid, LipO and phenolic compounds and at least one protein;
b) contacting the aqueous phase with a mobile solubilized ligand at physico-chemical conditions allowing formation of a complex between the ligand and the protein;
c) allowing the complex to separate from the aqueous supernatant phase, optionally by changing said physico-chemical conditions in the composition to reduce the solubility of the complex; and
d) isolating the one or more of glycoalkaloid, LipO and phenolic compounds comprised in the aqueous phase from the complex.
20. A method for isolating one or more of glycoalkaloid, LipO and phenolic compounds said method comprising:
a) providing an aqueous phase comprising one or more of glycoalkaloid, LipO and phenolic compounds and at least one protein;
b) contacting the aqueous phase with a mobile solubilized ligand at physico-chemical conditions allowing formation of a complex between the ligand and the one or more glycoalkaloid, LipO and phenolic compounds;
c) allowing the complex to separate from the aqueous supernatant phase, optionally by changing said physico-chemical conditions in the composition to reduce the solubility of the complex; and
d) isolating the one or more of glycoalkaloid, LipO and phenolic compounds comprised in the aqueous phase from the complex.