# US 10,765,135 B2 · Production of protein-polysaccharide conjugates > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,765,135 B2 - Document ID: US10765135B2 - Status: Granted - Kind: B2 - Application number: 12/407,543 - Filed: 2009-03-19 - Published: 2009-12-17 - Granted: 2020-09-08 - Earliest claimed: 2008-03-19 - Assignees: Wisconsin Alumni Research Foundation - Applicants: John A. Lucey; Dani Zhu; Srinivasan Damodaran - Inventors: John A. Lucey; Dani Zhu; Srinivasan Damodaran - Web page: https://patentgrep.com/patent/US10765135B2 ## CPC - A23L 29/30: containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin (products from apiculture A23L21/20; artificial sweetening agents A23L27/30) - A23C 21/08: containing other organic additives, e.g. vegetable or animal products - A23J 3/08: Dairy proteins - A23L 2/66: Proteins - A23L 5/00: Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor - A23L 27/60: Salad dressings; Mayonnaise; Ketchup - A23L 27/84: {Flavour masking or reducing agents} - A23L 29/10: containing emulsifiers - A23L 33/125: containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates (indigestible substances A23L33/21) - A61K 35/20: Milk; Whey; Colostrum - C07K 14/4717: {Plasma globulins, lactoglobulin} - C08B 37/0021: {Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran} - C08H 1/00: Macromolecular products derived from proteins (food proteins A23; glue, gelatine C09H) - A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - A23C: DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF (preservation thereof A23B11/00) - A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C08B: POLYSACCHARIDES; DERIVATIVES THEREOF (polysaccharides containing less than six saccharide radicals attached to each other by glycosidic linkages C07H; fermentation or enzyme-using processes C12P19/00; sugar industry C13; production of cellulose D21) - C08H: DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS (polysaccharides C08B; natural rubber C08C; natural resins or their derivatives C09F; bituminous materials C10) - A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION ## Abstract The present invention provides novel compositions and methods for producing protein-polysaccharide conjugates in aqueous solutions. Also provided are methods for limiting the Maillard reaction to the very initial stage, the formation of the Schiff base. Provided are methods to obtain a simple product of Schiff base with white color, and compositions obtained using the methods of the present invention. ## Claims ### Claim 1 A method of preparing a polysaccharide-protein conjugate, comprising reacting a polysaccharide comprising a reducing sugar and a protein in an aqueous solution comprising 10-40% (w/v) polysaccharide and 1-30% (w/v) protein, under temperature conditions of from about 40° C. to about 120° C., thereby producing a polysaccharide-protein conjugate, wherein the polysaccharide-protein conjugate is substantially free of intermediate and advanced Maillard products and exhibits a maximum absorbance peak at a wavelength of less than 420 mm in difference ultraviolet spectroscopy scanned between 270 nm and 500 nm. ### Claim 2 (depends on 1) The method of claim 1, wherein the reacting is conducted at a pH of from about 6.0 to about 8.0. ### Claim 3 (depends on 1) The method of claim 1, wherein the reducing sugar and protein are reacted under temperature conditions of from about 40° C. to about 120° C. for a period of from about 1 hour to about 48 hours. ### Claim 4 (depends on 1) The method of claim 1, further comprising subjecting the solution to a hydrostatic pressure in the amount of 1-20 MPa. ### Claim 5 (depends on 1) The method of claim 1 further comprising the step of recovering the polysaccharide-protein conjugate from the solution. ### Claim 6 A method of preparing a whey protein isolate-dextran conjugate, comprising reacting whey protein isolate and dextran in an aqueous solution comprising 10-40% (w/v) dextran and 1-30% (w/v) whey protein isolate, under temperature conditions of from about 40° C. to about 120° C., thereby producing whey protein isolate-dextran conjugate. ### Claim 7 (depends on 6) The method of claim 6, wherein the solution is acidified to a pH of from about 6.0 to about 8.0. ### Claim 8 (depends on 6) The method of claim 6, wherein the dextran and the whey protein isolate are reacted under temperature conditions of from about 40° C. to about 120° C. for a period of from about 1 hour to about 48 hours. ### Claim 9 (depends on 6) The method of claim 6, further comprising subjecting the solution to a hydrostatic pressure in the amount of 1-20 MPa. ### Claim 10 (depends on 6) The method of claim 6 further comprising the step of recovering the whey protein isolate-dextran conjugate from the solution. ### Claim 11 (depends on 1) The method of claim 1, wherein an amount by weight of the polysaccharide in the aqueous solution is greater than an amount by weight of the protein in the aqueous solution. ### Claim 12 (depends on 1) The method of claim 1, wherein the polysaccharide comprises at least one of galactomannan, maltodextrin, konjac mannan, xyloglucan, polydextrose, and dextrin. ### Claim 13 (depends on 1) The method of claim 1, wherein the polysaccharide comprises maltodextrin. ### Claim 14 (depends on 1) The method of claim 1, wherein the reducing sugar and protein are reacted under temperature conditions of from 40° C. to 70° C. for a period of from about 4 hours to less than 48 hours. ### Claim 15 (depends on 1) The method of claim 1, wherein the reducing sugar and protein are reacted under temperature conditions of from 40° C. to 70° C. for a period of from about 4 hours to about 24 hours. ### Claim 16 (depends on 1) The method of claim 1, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of less than 355 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 17 (depends on 1) The method of claim 1, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of less than 310 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 18 (depends on 1) The method of claim 1, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of about 304-305 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 19 (depends on 1) The method of claim 1, further comprising, after the reacting, cooling the aqueous solution to a temperature below the temperature conditions to thereby yield the polysaccharide-protein conjugate. ### Claim 20 (depends on 19) The method of claim 19, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of less than 355 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 21 (depends on 19) The method of claim 19, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of less than 310 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 22 (depends on 19) The method of claim 19, wherein the polysaccharide-protein conjugate exhibits a maximum absorbance peak at a wavelength of about 304-305 nm in difference ultraviolet spectroscopy scanned between wavelengths 270 nm and 500 nm. ### Claim 23 (depends on 19) The method of claim 19, wherein the reducing sugar and protein are reacted under temperature conditions of from 40° C. to 70° C. for a period of from about 4 hours to less than 48 hours. ### Claim 24 (depends on 19) The method of claim 19, wherein the reducing sugar and protein are reacted under temperature conditions of from 40° C. to 70° C. for a period of from about 4 hours to about 24 hours. ### Claim 25 (depends on 1) The method of claim 1, wherein the polysaccharide-protein conjugate is white in color.