# US 11,896,030 B2 · Tobacco-derived protein compositions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,896,030 B2 - Document ID: US11896030B2 - Status: Granted - Kind: B2 - Application number: 14/878,561 - Filed: 2015-10-08 - Published: 2016-02-04 - Granted: 2024-02-13 - Earliest claimed: 2013-05-17 - Assignees: R.J. Reynolds Tobacco Company - Applicants: R.J. Reynolds Tobacco Company - Inventors: Anthony Richard Gerardi; Crystal Dawn Hege Byrd; Thaddeus Jude Jackson; Chelsea Allison Cooke; John-Paul Mua; Kyle Ford - Web page: https://patentgrep.com/patent/US11896030B2 ## CPC - A23J 1/007: {from leafy vegetables, e.g. alfalfa, clover, grass} - A23J 3/14: Vegetable proteins - A23K 20/147: Polymeric derivatives, e.g. peptides or proteins - A23L 2/66: Proteins - A23L 5/23: {by extraction with solvents} - A23L 5/273: {using adsorption or absorption agents, resins, synthetic polymers, or ion exchangers} - A23L 33/185: Vegetable proteins - A24B 15/18: Treatment of tobacco products or tobacco substitutes - A24B 15/20: Biochemical treatment - A24B 15/24: by extraction; Tobacco extracts - A24B 15/241: {Extraction of specific substances} - C07K 14/415: from plants - C12N 9/88: Lyases (4.) - C12Y 401/01039: Ribulose-bisphosphate carboxylase (4.1.1.39) - A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs - A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS - A23K: FODDER - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - A24B: MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES - 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 disclosure describes methods for the purification of protein-enriched extracts to provide concentrates and isolates and methods for incorporation of such materials into products. The purification methods are adapted for removal of one or more of ash, metal salts, alkaloids, particulates, heavy metals, and other impurities and/or contaminants from extracts, as well as modifying the sensory characteristics (e.g., odor, color, and/or taste characteristics) of extracts. The methods generally include diafiltration, treatment with functionalized resins, and supercritical extraction. A protein composition in the form of a concentrate or isolate is provided, the protein composition including RuBisCO, F2 fraction proteins, or combination thereof extracted from a plant of the Nicotiana species, wherein the protein composition is characterized by one or more of: an ash content of less than about 15% by weight; a nicotine content of less than about 10 μg/g, and a heavy metal content of less than about 60 μg/g. ## Claims ### Claim 1 A method for modifying the content of a plant-derived, protein-enriched material, the method comprising receiving a plant-derived, protein-enriched material comprising RuBisCO, F2 fraction proteins, or a combination thereof, wherein the plant-derived, protein-enriched material further comprises one or more heavy metals; dissolving at least a portion of the plant-derived, protein-enriched material in a solvent to form a solution, contacting the plant-derived, protein-enriched material solution with a functionalized resin to afford a treated solution comprising a protein concentrate or isolate having a reduced amount of one or more of copper, cadmium, lead, and nickel as compared with the plant-derived, protein-enriched material, wherein the functionalized resin comprises a first resin functionalized with triaminetetraacetic acid chelating groups, and a second resin functionalized with triaminetetraacetate, sodium salt chelating groups; separating the treated solution from the functionalized resin, wherein the contacting step is conducted for a period of 1 to 3 hours; and wherein the contacting step comprises agitating the plant-derived, protein-enriched material solution with the functionalized resin at room temperature, and/or heating the plant-derived, protein-enriched material solution at a temperature of 50° C. to 70° C. ### Claim 2 (depends on 1) The method of claim 1, wherein the contacting step comprises stirring the plant-derived, protein-enriched material solution with the functionalized resin. ### Claim 3 (depends on 1) The method of claim 1, wherein the contacting step comprises passing the plant-derived, protein-enriched material solution through a column comprising the functionalized resin. ### Claim 4 (depends on 1) The method of claim 1, wherein the functionalized resin comprises a selective ion exchange resin. ### Claim 5 (depends on 1) The method of claim 1, wherein the functionalized resin is a strong acid cation resin or a weak acid cation resin. ### Claim 6 (depends on 1) The method of claim 1, wherein the solvent is an aqueous solvent. ### Claim 7 (depends on 1) The method of claim 1, wherein the solvent is acidic. ### Claim 8 (depends on 1) The method of claim 1, wherein the solvent is basic. ### Claim 9 (depends on 1) The method of claim 1, wherein the treated solution comprises an amount of one or more of copper, cadmium, lead, and nickel that is reduced at least 25% as compared with the plant-derived, protein-enriched material. ### Claim 10 (depends on 1) The method of claim 1, wherein the treated solution comprises an amount of one or more of copper, cadmium, lead, and nickel that is reduced at least 50% as compared with the plant-derived, protein-enriched material. ### Claim 11 (depends on 1) The method of claim 1, wherein the contacting step is conducted for a period of time of at least 2 hours. ### Claim 12 (depends on 1) The method of claim 1, wherein the first resin comprises a silica gel resin functionalized with triaminetetraacetic acid chelating groups and the second resin comprises a silica gel resin functionalized with triaminetetraacetate, sodium salt chelating groups. ### Claim 13 (depends on 1) The method of claim 1, wherein the contacting step is conducted for a period of time of 3 hours. ### Claim 14 (depends on 1) The method of claim 1, wherein the contacting step comprises agitating the plant-derived, protein-enriched material solution with the functionalized resin at room temperature, and heating the plant-derived, protein-enriched material solution at a temperature of 50° C. to 70° C. ### Claim 15 (depends on 1) The method of claim 1, wherein the contacting step is conducted for a period of time of 3 hours, and wherein the contacting step comprises agitating the plant-derived, protein-enriched material solution with the functionalized resin at room temperature, and heating the plant-derived, protein-enriched material solution at a temperature of 50° C. to 70° C. ### Claim 16 A method for modifying the copper content of a plant-derived, protein-enriched material, the method comprising receiving a plant-derived, protein-enriched material comprising RuBisCO, F2 fraction proteins, or a combination thereof, wherein the plant-derived, protein-enriched material further comprises one or more heavy metals; dissolving at least a portion of the plant-derived, protein-enriched material in a solvent to form a solution, contacting the plant-derived, protein-enriched material solution with a functionalized resin to afford a treated solution comprising a protein concentrate or isolate having a reduced amount of copper as compared with the plant-derived, protein-enriched material, wherein the functionalized resin comprises a first resin functionalized with triaminetetraacetic acid chelating groups, and a second resin functionalized with triaminetetraacetate, sodium salt chelating groups; separating the treated solution from the functionalized resin, wherein the contacting step is conducted for a period of 1 to 3 hours; and wherein the contacting step comprises heating the plant-derived, protein-enriched material solution at a temperature of 50° C. to 70° C. ### Claim 17 (depends on 1) The method of claim 1, for modifying the cadmium content of a plant-derived, protein-enriched material, wherein the contacting comprises heating the plant-derived, protein-enriched material solution at a temperature of 50° C. to 70° C., and wherein the contacting affords a treated solution comprising a protein concentrate or isolate having a reduced amount of cadmium as compared with the plant-derived, protein-enriched material. ### Claim 18 (depends on 1) The method of claim 1, for modifying the lead content of a plant-derived, protein-enriched material, wherein the contacting affords a treated solution comprising a protein concentrate or isolate having a reduced amount of lead as compared with the plant-derived, protein-enriched material. ### Claim 19 (depends on 1) The method of claim 1, for modifying the nickel content of a plant-derived, protein-enriched material, wherein the contacting comprises agitating the plant-derived, protein-enriched material solution with the functionalized resin at room temperature; and wherein the contacting affords a treated solution comprising a protein concentrate or isolate having a reduced amount of nickel as compared with the plant-derived, protein-enriched material. ### Claim 20 (depends on 1) The method of claim 1, wherein the plant-derived, protein-enriched material comprises at least 40% by weight of RuBisCO, F2 fraction proteins, or a combination thereof.