Protein-enriched tobacco-derived composition
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.
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 60% protein by dry weight,
wherein the treating step comprises adjusting the pH of the clarified protein-containing extract to a pH of between about 3 to about 4.5;
isolating a precipitate from the acidic extract; and
washing the precipitate to provide a RuBisCO-containing and F2 protein-containing protein-enriched material.
2. The method of claim 1 , wherein the washing is conducted on a filter having a pore size of between about 1 μm and about 1 kDa.
3. The method of claim 1 , wherein the solvent used to wash the precipitate is an acidic solution.
4. The method of claim 1 , wherein the protein-enriched material comprises at least 80% protein by dry weight.
5. 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.
6. The method of claim 1 , further comprising spray drying, freeze drying, or otherwise dehydrating the protein-enriched material.
7. 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.
8. The method of claim 7 , 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.
9. The method of claim 1 , wherein the solvent employed in the contacting step comprises a buffer solution.
10. The method of claim 1 , wherein the clarifying step comprises passing the liquid protein-containing extract through a filter press or into a centrifuge.
11. The method of claim 1 , wherein the at least 60% protein by dry weight comprises at least 50% by dry weight of RuBisCO.
12. The method of claim 1 , wherein the at least 60% protein by dry weight comprises at least 10% by dry weight of F2 protein.
13. The method of claim 1 , further comprising separating the RuBisCO-containing and F2 protein-containing protein-enriched material to separately provide a RuBisCO-enriched material and an F2 protein-enriched material.
14. The method of claim 12 , wherein the separating step comprises dissolving the RuBisCO-containing and F2 protein-containing protein-enriched material in a solvent to give a mixture and bringing the mixture into contact with one or more filters or membranes to give a retentate comprising RuBisCO and a permeate comprising F2 protein.
15. The method of claim 13 , wherein the filters or membranes comprise a 500 kDa ultrafiltration membrane or a 0.1 μm ceramic filter.
16. The method of claim 1 , wherein the clarifying and treating steps are conducted at ambient temperature.