Methods for extracting and purifying non-denatured proteins

Patent metadata

Field	Value
Publication number	US-10087434-B2
Application number	US-201715661873-A
Country	US
Kind code	B2
Filing date	Jul 27, 2017
Priority date	Oct 1, 2014
Publication date	Oct 2, 2018
Grant date	Oct 2, 2018

Abstract

Materials and methods for extracting and purifying proteins are provided. For example, the materials and methods provided herein can be used for extracting and purifying proteins that denature at low temperature.

First claim

What is claimed is:

1. A method for purifying a pea albumin protein, comprising:
extracting a disintegrated pea biomass to remove solids and generate a protein suspension;
adding salt to the protein suspension;
adjusting the pH of the protein suspension to a pH of 2 to 10;
dialyzing the protein suspension against a polyethylene glycol (PEG) solution, or subjecting the protein suspension to ultrafiltration using a permeable membrane to separate the protein suspension on one side of the membrane from a permeate on the other side of the membrane, wherein PEG is present on the permeate side of the membrane; and
subjecting the dialyzed or ultrafiltered protein solution to one or more concentrating or filtering steps to generate a product phase that contains the pea albumin protein.

2. The method of claim 1 , wherein the pea biomass comprises pea seeds.

3. The method of claim 1 , wherein the extracting comprises using gravity settling or centrifugation to separate solids from the protein suspension.

4. The method of claim 1 , further comprising adding a flocculant to the pea biomass.

5. The method of claim 4 , wherein the flocculant comprises alkylamine epichlorohydrin, polydimethyldiallylammonium chloride, or a polyamine.

6. The method of claim 4 , wherein the flocculant comprises alkylamine epichlorohydrin.

7. The method of claim 1 , wherein the PEG has a molecular weight of about 3000 to about 10,000.

8. The method of claim 1 , wherein the PEG has a molecular weight of about 8000.

9. The method of claim 1 , wherein the PEG is present at a volume concentration of 2% to 50%.

10. The method of claim 1 , wherein the salt comprises sodium chloride, magnesium sulfate, or a mixture thereof.

11. The method of claim 1 , wherein the salt is added to the protein suspension to attain a concentration of about 100 mM to about 2M.

12. The method of claim 1 , wherein the step of subjecting the dialyzed or ultrafiltered protein solution to one or more concentrating or filtering steps comprises diafiltering the dialyzed or ultrafiltered protein solution.

13. The method of claim 1 , wherein the step of subjecting the dialyzed or ultrafiltered protein solution to one or more concentrating or filtering steps comprises concentrating and filtering the dialyzed or ultrafiltered protein solution to generate a pea albumin protein concentrate.

14. The method of claim 13 , further comprising sterilizing the pea albumin protein concentrate to obtain a sterilized pea albumin protein concentrate, and drying the sterilized pea albumin protein concentrate.

15. The method of claim 1 , comprising subjecting the protein suspension to ultrafiltration using a permeable membrane, wherein the membrane comprises polyethersulfone, polypropylene, polyvinylidene fluoride, polyacrylonitrile, cellulose acetate, or polysulfone.

Assignees

• Impossible Foods Inc

Inventors

• Kale Aniket
• Varadan Ranjani
• Davis Simon Christopher

Classifications

• B01D61/145

  Ultrafiltration · CPC title

• C12N9/88 Primary

  Lyases (4.) · CPC title

• B01D11/02

  of solids · CPC title

• A23J1/006

  from vegetable materials (A23J1/005, A23J1/12 and A23J1/14 take precedence) · CPC title

• C12Y401/01039

  Ribulose-bisphosphate carboxylase (4.1.1.39) · CPC title