CA2866977C – March 19, 2019 – AQUEOUS PROCESS FOR PREPARING PROTEIN ISOLATE AND HYDROLYZED PROTEIN FROM AN OILSEED

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Inventors :

ROZENSZAIN, LUIS; BEYE, GARRISON

Owner :

A23J1 / 14; A23J3 / 16; C07K1 / 14; C07K1 / 34; C11B1 / 10; C12P13 / 04; C12P21 / 06

Application Number :

CA2866977

Document Number :

CA2866977C

Priority Date :

January 1, 1970

Filing Date :

April 4, 2012

Date of Grant/ Publication :

March 19, 2019

Class :

SIEBTE PMI VERWALTUNGS GMBH

Abstract

The present disclosure relates to an aqueous process for the preparation of a protein isolate and a hydrolyzed protein concentrate from an oilseed meal, optionally comprising: mixing an oilseed meal with an aqueous solvent to form a slurry; optionally treating the slurry with phytase y; separating the slurry with a solid/liquid separation to form: a liquid phase, comprising the aqueous solvent, soluble protein and oil; and a solid phase comprising insoluble protein; separating the liquid phase to form: an oil phase; and an aqueous protein phase; subjecting the aqueous protein phase to membrane filtration to obtain a protein solution; and drying the protein solution to obtain the protein isolate subjecting the insoluble protein to enzymatic hydrolysis, and subjecting the hydrolyzed protein to membrane filtration to obtain an amino acid and peptide solution; and drying the amino acid and peptide solution to obtain the hydrolyzed protein concentrate.

Claim(s)

1. A process for producing a protein isolate from an oilseed meal, comprising: providing an oil seed meal; mixing an oilseed meal with an aqueous solvent to form a slurry; optionally treating the slurry with phytase at a temperature and a pH suitable for phytase activity; separating the slurry with a solid/liquid separation to form: a liquid phase, comprising the aqueous solvent, soluble protein and oil; and a solid phase; separating the liquid phase to form: an oil phase; and an aqueous protein phase; subjecting the aqueous protein phase to membrane filtration to obtain a protein solution; and drying the protein solution to obtain the protein isolate.;
2. The process according to claim 1, wherein the process further comprises mixing the solid phase with an aqueous solvent and repeating the solid/liquid separation from one to four times.;
3. The process according to claim 1 or 2, wherein the ratio of the oilseed meal to the aqueous solvent is between 1:6 to 1:15 (w/w) of meal to solvent.;
4. The process according to claim 3, wherein the ratio of the oilseed meal to the aqueous solvent is 1:8 to 1:12.5 (w/w) of meal to solvent.;
5. The process according to any one of claims 1 to 4, wherein the temperature suitable for phytase activity is between 20° and 65°C and the pH suitable for phytase activity is between 4.5 and 7.5.;
6. The process according to claim 5, wherein the temperature suitable for phytase activity is 50°C-55°C and the pH suitable for phytase activity is between 5.8-6.2.;
7. The process according to any one of claim 1 to 6, wherein the solid liquid separation is conducted by centrifugation.;
8. The process according to claim 7, wherein the solid liquid separation of the mixture is conducted by a decanter centrifuge.;
9. The process according to claim 8, wherein the decanter centrifuge is operated at a g-force between 2,000-4,000g.;
10. The process according to claim 9, wherein the g-force is 3,000g.;
11. The process according to claim 7, wherein the centrifugation comprises a decanter centrifuge followed by a first and second disk stack centrifuge in series.;
12. The process according to claim 11, wherein the first disk stack centrifuge is operated at a g-force between 6,000-9,000g, and the second disk stack centrifuge is operated at a g-force between 8,000g-12,000g.;
13. The process according to claim 12, wherein the first disk stack centrifuge is operated at a g-force of 8,300g, and the second disk stack centrifuge is operated at a g-force of 10,000g.;
14. The process according to any one of claim 1 to 13, wherein the separation of the liquid phase is conducted by centrifugation.;
15. The process according to claim 14, wherein the separation of the liquid phase is conducted by a skimming centrifuge, a disk stack centrifuge, a 2-phase centrifuge or a 3-phase centrifuge or a combination thereof.;
16. The process according to claim 15, wherein the separation of the liquid phase is conducted by a disk stack centrifuge.;
17. The process according to claim 16, wherein the centrifuge is operated at a g-force between 2,000-10,000g.;
18. The process according to claim 17, wherein the centrifuge is operated at a g-force of 6,000g.;
19. The process according to any one of claims 1 to 18, wherein the aqueous protein phase is subjected to ultrafiltration followed by diafiltration to obtain a retentate suitable for spray drying comprising the protein solution.;
20. The process according to any one of claims 1 to 19, wherein the aqueous solvent comprises water.;
claim 21: 21. The process according to claim 20 wherein the aqueous solvent consists of water.;
claim 22: 22. The process according to any one of claims 1 to 21, wherein the protein isolate comprises less than 2% (w/w) of oil.;
claim 23: 23. The process according to any one of claims 1 to 22, wherein the protein isolate comprises at least 90% protein (w/w) on a dry weight basis.;
claim 24: 24. The process according to any one of claims 1 to 23, wherein the oilseed meal comprises a canola, rapeseed, mustard seed, broccoli seed, flax seed, cotton seed, hemp seed, safflower seed, sesame seed or soybean meal.;
claim 25: 25. The process according to claim 24, wherein the oilseed meal comprises canola meal.;
claim 26: 26. The process according to any one of claims 1 to 25, wherein the process is a continuous process.;
claim 27: 27. The process according to any one of claims 1 to 26, wherein the solid phase is dried and solvent extracted to isolate oil.;
claim 28: 28. The process according to any one of claims 1 to 27, wherein the slurry is further treated with cellulose.;
claim 29: 29. The process according to any one of claims 1 to 28, wherein membrane filtration comprises one or more steps of microfiltration, ultrafiltration and diafiltration.;
claim 30: 30. The process according to any one of claims 1 to 29, wherein the oil seed meal is provided by cold pressing.;
claim 31: 31. The process according to any one of claims 1 to 30 where in the temperature in the cold pressing does not exceed 85°C.;
claim 32: 32. A protein isolate prepared according to the process of any one of claims 1 to 32.;
claim 33: 33. A protein isolate having substantially the same amount of soluble protein as in the oil seed or pressed cake from which the isolate is recovered.;
claim 34: 34. A protein isolate having an amino acid distribution as listed in Table 19.;
claim 35: 35. A protein isolate having a molecular weight distribution as listed in Table 21.;
claim 36: 36. A protein isolate comprising: i) a first portion of proteins having a molecular weight of above about 300 kDa, wherein the first portion represents between 40 and 65% of the total protein: ii) a second portion of proteins having a molecular weight of between about 30-10 kDa, wherein the second portion represents between 10 and 40% of the total protein; and ?) a third portion of proteins having a molecular weight of between about 10-5 kDa, wherein the third portion represents between 5 and 30% of the total protein.;
claim 37: 37. A protein isolate prepared according to the process of any one of claims 1 to 31 having a soluble protein concentration of at least 90%.;
claim 38: 38. A process for producing soluble amino acids and peptides from a composition comprising insoluble proteins comprising: providing an oil seed meal; mixing the oilseed meal with an aqueous solvent to form a slurry; optionally treating the slurry with phytase at a temperature and a pH suitable for phytase activity; separating the slurry using solid/liquid separation procedures to form: a liquid phase, comprising the aqueous solvent, soluble protein and oil; and a solid phase, comprising residual aqueous solvent, insoluble protein and oil washing the solid phase with water and subjecting the washed solid phase to centrifugation to form: a liquid extract; and an insoluble protein phase; dispersing the insoluble protein phase in water to form an aqueous dispersion of insoluble protein, and adjusting the temperature and pH of the aqueous dispersion to a condition suitable for enzymatic action of one or more of an endopeptidases or protease, adding a first endopeptidases or protease enzyme to the temperature and pH adjusted dispersion of insoluble protein, stirring the enzyme-containing dispersion of insoluble protein for a predetermined period of time to form hydrolyzed protein in said dispersion, and subjecting the hydrolyzed protein dispersion to a separation process to obtain: a solution of soluble amino acids and peptides; a solid phase comprising insoluble protein and an oil.;
claim 39: 39. The process of claim 38 further including drying the solution of hydrolyzed soluble protein and peptides to obtain a dry soluble protein and peptide composition substantially free of water and oil.;
claim 40: 40. The process of claim 38 or 39, wherein the oil seed meal comprises a dry, partially defatted pressed cake meal, or a wet oily meal, wherein the oil seed meal has at least about 9? oil.;
claim 41: 41. The process according to any one of claims 38 to 40, wherein the process further comprises mixing the solid phase with water and repeating a solid/liquid separation from one to four times.;
claim 42: 42. The process of any one of claims 38 to 41, wherein the temperature suitable for enzyme activity is 60°C and the suitable pH is about 8.3.;
claim 43: 43. The process according to claim 42 wherein a first enzyme is an endopeptidase and the endopeptidase is stirred with the dispersion of insoluble protein.;
claim 44: 44. The process according to claim 43 wherein the endopeptidase is stirred with the dispersion of insoluble protein for about 4 hours.;
claim 45: 45. The process according to claim 43 or 44 wherein, following stirring, the temperature of the enzyme containing slurry is reduced to about 50°C, a second endopeptidase or exopeptidase or protease is added to the enzyme containing slurry and said slurry is stirred.;
claim 46: 46. The process of claim 45 wherein the stirring of the slurry is continued for about an additional four hours.;
claim 47: 47. The process according to any one of claims 38 to 46, wherein the separation process comprises one or more filtration procedures including one or more of ultrafiltration, nanofiltration and diafiltration.;
claim 48: 48. The process according to claim 47, wherein the separation process comprises one or more filtration procedures including one or more of ultrafiltration, nanofiltration and diafiltration.;
claim 49: 49. The process according to any one of claims 38 to 48 further including contacting the solution of soluble amino acids and peptides with carbon prior to a filtration step.;
claim 50: 50. The process of any one of claims 38 to 49, wherein the oil seed meal is subjected to a process to remove fiber prior to mixing the oilseed meal with the aqueous solvent to form the slurry.;
claim 51: 51. The process of any one of claims 38 to 50 wherein the solution of soluble amino acids and peptides is subjected to nanofiltration to obtain a retentate suitable for spray drying.;
claim 52: 52. The process according to any one of claims 38 to 51, wherein the soluble amino acids and peptides are provided as dry hydrolysate products having from about 79% to about 95% protein on a dry weight basis.;
claim 53: 53. The process of claim 52 wherein the dry hydrolyzates have an amino acid range as set forth in the following table: Range Amino acid {g./100g product) Aspartic acid 6-8 Glutamic acid 15-20 Serine 3-5 Glycine 4-5 Histidine 2-4 Arginine 3-6 Threonine 3-4 Alanine 3-5 Proline 5-6 Tyrosine 2-4 Valine 4-6 Methionine 2-3 Cystine 2-3 isoleucine 3-5 Leucine 6-8 Phenylalanine 3-5 Lysine 4-6 Tryptophan 1-2.;
claim 54: 54. The process of claim 53 wherein the dry hydrolyzates have an amino acid profile as set forth in the following table:;
claim 55: 55. The process of any one of claims 52 to 54 wherein the dry hydrolyzates have a amino acid and peptide molecular weight profile as set forth in the following table:;
claim 56: 56. The process of claim 52, wherein the dry hydrolyzates have a peptide molecular weight profile comprising: i) a first portion of hydrolyzed protein monomers, wherein the first portion represents between 25 and 50% of the total protein; ?) a second portion of hydrolyzed protein dimers, wherein the second portion represents between 35 and 65% of the total protein; and iii) a third portion of hydrolyzed protein having a molecular weight of between about 3-0.6 kDa, wherein the third portion represents between 5 and 25% of the total protein; and optionally iv) a fourth portion of hydrolyzed protein having a molecular weight of between 3-5 kDa, wherein the fourth portion represents between 0 and 5% of the total protein.;
claim 57: 57. The process of any one of claims 38 to 56, wherein separation process comprises one or more steps of microfiltration, ultrafiltration and diafiltration.;
claim 58: 58. The process of any one of claims 38 to 57, wherein the oil seed meal is provided by cold pressing.;
claim 59: 59. The process of any one of claims 38 to 58, wherein the endopeptidase is provided at a concentration of up to about 1.5% of the oil seed meal and the exopeptidase or protease is provided at a concentration up to about 0.4% of the oil seed meal;
claim 60: 60. Dry hydrolysate products derived from an oily pressed canola meal, said dry hydrolysate products having from about 79% to about 95% amino acid and peptides on a dry weight basis and amino acid profiles as set forth in the following table;;
claim 61: 61. Dry hydrolysate products derived from an oily pressed canola meal as claimed in claim 60, said dry hydrolysate products having from about 79% to about 95% amino acid and peptides on a dry weight basis and amino acid profiles as set forth in the following table;
claim 62: 62. The dry hydrolysate products of claim 60 or 61 wherein the dry hydrolyzates have an amino acid and peptide molecular weight profile as set forth in the following table:;
claim 63: 63. The dry hydrolysate products of claim 60 wherein the dry hydrolyzates have a peptide molecular weight profile comprising: i) a first portion of hydrolyzed protein monomers, wherein the first portion represents between 25 and 50% of the total protein; ii) a second portion of hydrolyzed protein dimers, wherein the second portion represents between 35 and 65% of the total protein; and iii) a third portion of hydrolyzed protein having a molecular weight of between about 3-0.6 kDa, wherein the third portion represents between 5 and 25% of the total protein; and optionally iv) a fourth portion of hydrolyzed protein having a molecular weight of between 3-5 kDa, wherein the fourth portion represents between 0 and 5% of the total protein.;
claim 64: 64. A process for forming a dry soluble amino acid and peptide mixture from a pressed cake of canola meal, said pressed cake containing at least about 9% oil comprising a) separating insoluble proteins in said pressed cake from the fiber, soluble proteins, and oil therein, b) forming a slurry of the insoluble proteins in water, c) adjusting the temperature and pH of said slurry to a condition suitable for enzymatic activity of a first hydrolyzing enzyme, reacting said slurry with said first hydrolyzing enzyme for a preset period of time, d) adjusting the temperature and pH of said slurry to a condition suitable for enzymatic activity of a second hydrolyzing enzyme, reacting said slurry with said second hydrolyzing enzyme for a preset period, e) the first and second hydrolyzing enzymes reacting with the insoluble proteins to form soluble amino acids and peptides, f) separating the soluble amino acids and peptides from residual insoluble materials, and g) drying the separated soluble amino acids and peptides

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