Purification of tobacco-derived protein compositions
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 nicotine and may have other benefits, e.g., lightening the color of the protein-enriched extracts. The methods generally include treatment with peracetic acid or hydrogen peroxide and filtrations. 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 relatively low nicotine content.
1. A method for reducing the nicotine content in a plant-derived protein-enriched material, the method comprising:
a) obtaining a plant-derived, protein-enriched material comprising RuBisCO, F2 fraction proteins, or a combination thereof, wherein the plant-derived, protein-enriched material comprises nicotine;
b) treating the plant-derived, protein-enriched material with peracetic acid to give a peracetic acid-treated mixture;
c) washing the peracetic acid-treated mixture with water on a filter, wherein a solid treated protein-containing material is retained on the filter;
d) solubilizing the solid treated protein-containing material to give a solution and adjusting the pH of the solution to a basic pH to give a basic solution; and
e) processing the basic solution on a filter to provide a retentate comprising a protein concentrate or isolate having a reduced amount of nicotine compared with the plant-derived, protein-enriched material.
2. The method of claim 1 , wherein the plant-derived, protein-enriched material comprises material from a plant of the Nicotiana species.
3. The method of claim 1 , wherein the treating step b) is conducted at a pH of about 3 to about 5.
4. The method of claim 1 , wherein the treating step b) is conducted at a temperature of about 25° C. to about 42° C.
5. The method of claim 1 , wherein the treating step b) comprises treating the plant-derived, protein-enriched material with peracetic acid in the form of an aqueous solution comprising at least about 3% by weight peracetic acid.
6. The method of claim 1 , wherein step c) comprises washing the peracetic acid-treated mixture with water on a 1.4 μm pore filter.
7. The method of claim 1 , wherein step c) comprises washing the peracetic acid-treated mixture with water having a pH of less than about 5.
8. The method of claim 1 , wherein step d) comprises resolubilizing the solid treated protein-containing material in an aqueous buffer.
9. The method of claim 8 , wherein the aqueous buffer comprises sodium bicarbonate and potassium carbonate.
10. The method of claim 1 , wherein step e) comprises processing the basic solution on a 10 nm filter.
11. The method of claim 1 , further comprising spray drying the protein concentrate or isolate.
12. The method of claim 1 , wherein the protein concentrate or isolate comprises about 5 ppm nicotine or less.
13. The method of claim 1 , wherein the protein concentrate or isolate comprises about 80% or more RuBisCO, F2 fraction proteins, or a combination thereof by weight.