# US 10,945,448 B2 · Process for removing phospholipids and off-flavors from proteins and resulting protein product > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,945,448 B2 - Document ID: US10945448B2 - Status: Granted - Kind: B2 - Application number: 12/861,077 - Filed: 2010-08-23 - Published: 2011-02-24 - Granted: 2021-03-16 - Earliest claimed: 2009-08-21 - Assignees: Wisconsin Alumni Research Foundation - Applicants: Srinivasan Damodaran; Akshay Arora - Inventors: Srinivasan Damodaran; Akshay Arora - Web page: https://patentgrep.com/patent/US10945448B2 ## CPC - A23J 3/16: from soybean - A23L 5/273: {using adsorption or absorption agents, resins, synthetic polymers, or ion exchangers} - A23L 11/34: {using chemical treatment, adsorption or absorption} - A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) ## Abstract Described are methods of removing phospholipids and other off-flavor-causing compounds from edible proteins using a cyclodextrin treatment. The methods include treating soy protein with cyclodextrins such as β-cyclodextrin to form cyclodextrin-compound complexes and then separating the resulting complexes from the protein. Optionally, prior to treating the protein with cyclodextrin, the protein is sonicated and then treated with a phospholipase, such as phospholipase A2. Versions of the methods described herein are capable of removing more than 99% of phospholipids from soy protein. ## Claims ### Claim 1 A method for removing compounds that cause off-flavors in proteins, the method comprising: treating a protein-containing solution comprising a phospholipid with a phospholipase to yield a hydrolytic byproduct of the phospholipid in the solution; contacting the protein-containing solution with a cyclodextrin for a time wherein the cyclodextrin binds to the hydrolytic byproduct, thereby yielding a complex comprising the cyclodextrin bound to the hydrolytic byproduct; and then separating the complex from the protein-containing solution. ### Claim 2 (depends on 1) The method of claim 1 wherein the protein-containing solution is derived from oilseeds or oil-bearing cereal grain. ### Claim 3 (depends on 1) The method of claim 1 wherein the contacting step includes contacting the protein-containing solution with an unsubstituted or substituted β-cyclodextrin. ### Claim 4 (depends on 1) The method of claim 1 wherein the cyclodextrin in the contacting step is immobilized on a solid support. ### Claim 5 (depends on 1) The method of claim 1 wherein the treating step includes treating the protein-containing solution with a phospholipase A2. ### Claim 6 (depends on 1) The method of claim 1 wherein the treating step includes treating the protein-containing solution with a phospholipase A2 at a temperature of from about 20° C. to about 50° C. and at a concentration of from about 50 units to about 200 units phospholipase A2 per gram of protein in the protein-containing solution. ### Claim 7 (depends on 1) The method of claim 1 wherein the hydrolytic byproduct comprises a product selected from the group consisting of a fatty acid, a free fatty acid, an unsaturated fatty acid, and a lysophospholipid. ### Claim 8 (depends on 1) The method of claim 1 wherein the phospholipase is selected from the group consisting of a phospholipase A1, a phospholipase A2, a phospholipase B, a phospholipase C, and a phospholipase D. ### Claim 9 (depends on 1) The method of claim 1 wherein the protein-containing solution in the treating step comprises purified, defatted protein. ### Claim 10 A method for removing compounds that cause off-flavors in proteins, the method comprising: sonicating a protein-containing solution comprising a phospholipid; treating the protein-containing solution with a phospholipase to yield a hydrolytic byproduct of the phospholipid in the solution, wherein the sonicating is performed prior to the treating; contacting the protein-containing solution with a cyclodextrin for a time wherein the cyclodextrin binds to the hydrolytic byproduct, thereby yielding a complex comprising the cyclodextrin bound to the hydrolytic byproduct; and then separating the complex from the protein-containing solution. ### Claim 11 (depends on 10) The method of claim 10 wherein the sonicating step includes sonicating the protein-containing solution for at least about 2 minutes, at a temperature of from about 20° C. to about 70° C., and at a frequency of from about 10 kHz to about 30 kHz. ### Claim 12 (depends on 11) The method of claim 11 wherein the sonicating step includes sonicating the protein-containing solution for at least about 5 minutes. ### Claim 13 (depends on 1) The method of claim 1 wherein the separating step includes separating the complex from the protein-containing solution via ultrafiltration, dialysis, or precipitation. ### Claim 14 (depends on 1) The method of claim 1 wherein the separating step includes separating the complex from the protein-containing solution via isoelectric precipitation at a pH of from about 4 to 5. ### Claim 15 (depends on 1) The method of claim 1 wherein the separating step includes separating the complex from the protein-containing solution via ultrafiltration, wherein the ultrafiltration includes diafiltration. ### Claim 16 (depends on 15) The method of claim 15 wherein the diafiltration is performed with a diafiltration ratio of at least 2×. ### Claim 17 (depends on 15) The method of claim 15 wherein the diafiltration is performed with a diluent of water that includes a cyclodextrin. ### Claim 18 (depends on 10) The method of claim 10 wherein the protein-containing solution includes soy protein. ### Claim 19 (depends on 10) The method of claim 10 wherein the contacting step includes contacting the protein-containing solution with an unsubstituted or substituted β-cyclodextrin. ### Claim 20 (depends on 10) The method of claim 10 wherein the cyclodextrin is immobilized on a solid support. ### Claim 21 (depends on 10) The method of claim 10 wherein the treating step includes treating the protein-containing solution with a phospholipase A2 at a temperature of from about 20° C. to about 50° C. and at a concentration of from about 50 units to about 200 units phospholipase A2 per gram of protein in the protein-containing solution. ### Claim 22 (depends on 10) The method of claim 10 wherein the separating step includes separating the complex from the protein-containing solution via ultrafiltration using diafiltration at a diafiltration ratio of at least 2× or isoelectric precipitation at a pH of from about 4 to 5. ### Claim 23 (depends on 10) The method of claim 10 wherein the phospholipase is selected from the group consisting of a phospholipase A1, a phospholipase A2, a phospholipase B, a phospholipase C, and a phospholipase D. ### Claim 24 (depends on 10) The method of claim 10 wherein the protein-containing solution in the sonicating step comprises purified protein. ### Claim 25 (depends on 10) The method of claim 10 wherein the protein-containing solution in the sonicating step comprises purified, defatted protein. ### Claim 26 A method for removing compounds that cause off-flavors in proteins, the method comprising: treating a protein-containing solution comprising soy protein and a phospholipid with a phospholipase to yield a hydrolytic byproduct of the phospholipid in the solution; contacting the protein-containing solution with a cyclodextrin for a time wherein the cyclodextrin binds to the hydrolytic byproduct, thereby yielding a complex comprising the cyclodextrin bound to the hydrolytic byproduct; and then separating the complex from the protein-containing solution. ### Claim 27 (depends on 26) The method of claim 26 wherein the phospholipase is selected from the group consisting of a phospholipase A1, a phospholipase A2, a phospholipase B, a phospholipase C, and a phospholipase D.