PROTEIN-ENRICHED TOBACCO-DERIVED COMPOSITION

• US 20140271952A1
• Filed: 03/14/2013
• Published: 09/18/2014
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: R J Reynolds Tobacco Company (R. J. Reynolds Tobacco)
• Original Assignee: R J Reynolds Tobacco Company (R. J. Reynolds Tobacco)
• Inventors: Mua, John-Paul, Joyce, Leigh Ann Blevins, Lovette, Margarette Elisa, Morton, Joshua D., Ford, Kyle, Hagan, Leigh, Bratcher, Barry, Debusk, Samuel Mark
• Granted Patent: US 9,301,544 B2
• Time in Patent Office: Days
• US Class Current: 424/751
• CPC Class Codes: A23L 2/66 Proteins
• CPC Class Codes: A24B 15/24 by extraction; Tobacco extracts
• CPC Class Codes: A24B 15/241 Extraction of specific substances
• CPC Class Codes: A61K 2236/00 Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
• CPC Class Codes: A61K 2800/10 General cosmetic use
• CPC Class Codes: A61K 36/81 Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61K 8/9789 Magnoliopsida [dicotyledons]
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: C07K 1/145 by extraction or solubilisation
• CPC Class Codes: C07K 14/415 from plants

Abstract

The present disclosure describes methods of obtaining and/or deriving proteins from plants of the Nicotiana species and methods for incorporation of such proteins into various products. For example, a method for obtaining a protein-enriched material from a plant of the Nicotiana species or portion thereof is provided, comprising: extracting one or more proteins from the plant material into a solvent to form a liquid protein-containing extract; separating a solid extracted plant material from the liquid protein-containing extract; clarifying the liquid protein-containing extract to form a clarified protein-containing extract and a solids fraction; and treating the clarified protein-containing extract so as to provide a protein-enriched material comprising at least about 60% protein by weight.

Claims

1. A method for obtaining a protein-enriched material from a plant of the Nicotiana species or portion thereof, comprising:

receiving a plant material of the Nicotiana species;

contacting the plant material with a solvent for a time and under conditions sufficient to extract one or more proteins from the plant material into the solvent and form a liquid protein-containing extract;

separating a solid extracted plant material from the liquid protein-containing extract;

clarifying the liquid protein-containing extract to form a clarified protein-containing extract and a solids fraction; and

treating the clarified protein-containing extract so as to provide a protein-enriched material comprising at least about 60% protein by dry weight.

2. The method of claim 1, wherein the treating step comprises adjusting the pH of the clarified protein-containing extract to a pH of less than about 6 to form an acidic extract;

isolating a precipitate from the acidic extract; and

washing the precipitate to provide a protein-enriched material.

3. The method of claim 2, wherein the pH of the clarified protein-containing extract is adjusted to a pH of between about 4.5 to about 6 to provide a RuBisCO-containing protein-enriched material.

4. The method of claim 3, wherein the extract remaining after the isolating step is further treated by one or both of:

1) filtration on a filter having a pore size of 1 kDa-500 kDa and isolating a second protein-enriched material as a retentate; and

     2) adjusting the extract to a pH of less than about 4.5 and isolating a second precipitate comprising a second protein-enriched material therefrom, wherein the second protein-enriched material is an F2 protein-containing protein-enriched material.

5. The method of claim 2, wherein the washing is conducted on a filter having a pore size of between about 1 μ

m and about 1 kDa.

6. The method of claim 2, wherein the pH of the clarified protein-containing extract is adjusted to a pH of between about 3 to about 4.5 to provide a RuBisCO-containing and F2 protein-containing protein-enriched material.

7. The method of claim 2, wherein the solvent used to wash the precipitate is an acidic solution.

8. The method of claim 2, wherein the protein-enriched material comprises at least about 80% protein by dry weight.

9. The method of claim 1, wherein the treating step comprises filtering the clarified protein-containing extract on a ceramic filter or an ultrafiltration membrane to give a retentate and a liquid permeate;

and

washing the retentate to provide the protein-enriched material.

10. The method of claim 9, wherein the clarifying step comprises adjusting the pH of the liquid protein-containing extract.

11. The method of claim 9, wherein the filtering step comprises passing the clarified protein-containing extract through one or more filters having pore sizes of between about 1 μ

m and about 1 kDa.

12. The method of claim 9, wherein the filtering step comprises passing the clarified protein-containing extract through a filter having a pore size of between about 1 μ

m and about 500 kDa and wherein the retentate comprises at least about 50% RuBisCO by dry weight.

13. The method of claim 9, wherein the filtering step comprises passing the clarified protein-containing extract through a filter having a pore size of between about 1 kDa and about 500 kDa and wherein the retentate comprises a mixture of RuBisCO and F2 protein.

14. The method of claim 12, further comprising passing the permeate through a filter having a pore size of between about 500 kDa and 1 kDa to give a second protein-enriched material as the retentate, wherein the second protein-enriched material comprises F2 fraction proteins.

15. The method of claim 1, wherein plant material of the Nicotiana species is in the form of a green plant material, yellowed plant material, cured plant material, or a mixture thereof.

16. The method of claim 1, further comprising spray drying, freeze drying, or otherwise dehydrating the protein-enriched material.

17. The method of claim 1, further comprising adding one or more components to remove color, odor, taste, alkaloids, metals, or a combination thereof, at any step of the process.

18. The method of claim 17, wherein the one or more components are selected from the group consisting of activated carbon, a resin, clay, a chelating agent, a molecularly imprinted polymer, a non-imprinted polymer, or a combination thereof.

19. A protein-enriched extract obtained according to the method of claim 1.

20. A protein-enriched extract obtained according to the method of claim 2.

21. A protein-enriched extract obtained according to the method of claim 9.

22. A dietary supplement, food, beverage, personal care item, pharmaceutical product, or pet food comprising the protein-enriched extract of claim 19.

23. A dietary supplement, food, beverage, personal care item, pharmaceutical product, or pet food comprising the protein-enriched extract of claim 20.

24. A dietary supplement, food, beverage, personal care item, pharmaceutical product, or pet food comprising the protein-enriched extract of claim 21.