Method for producing a protein hydrolysate

Patent metadata

Field	Value
Publication number	US-10619177-B2
Application number	US-201515531944-A
Country	US
Kind code	B2
Filing date	Dec 1, 2015
Priority date	Dec 1, 2014
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

The present invention relates to a method of producing a protein hydrolysate comprising a step of enzymatic protein hydrolysis performed at high temperature.

First claim

The invention claimed is:

1. A method for producing a protein hydrolysate, comprising:
a) adding to a composition comprising a substrate protein a thermostable endopeptidase;
b) performing a first hydrolysis step by incubating the composition of step a) for at least 10 minutes at a temperature of at least 75° C.;
c) adding to the composition of step b) a protease preparation having an aminopeptidase activity of at least 200 LAPU/g; and
d) performing a second hydrolysis step by incubating the composition of step c) for at least 10 minutes at a temperature which is at least 10° C. lower than the temperature used in step b).

2. The method of claim 1 , wherein the thermostable endopeptidase is a nonspecific endopeptidase.

3. The method of claim 2 , wherein the nonspecific endopeptidase is characterized in that incubation of 0.5% (w/w) BSA with the endopeptidase for 4 hours at a temperature and pH where the endopeptidase exhibits at least 40% of its maximum activity results in a degree of hydrolysis of at least 10%.

4. The method of claim 1 , wherein the thermostable endopeptidase is an endopeptidase, which after incubation for 15 minutes at 80° C. and pH 9 has a residual activity of at least 80% relative to its activity after incubation at 37° C.

5. The method of claim 1 , wherein the thermostable endopeptidase (i) has at least 60% sequence identity to the polypeptide of SEQ ID NO: 3, (ii) is encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) is a variant of the polypeptide of SEQ ID NO: 3 comprising a substitution, deletion, and/or insertion at one or more positions.

6. The method of claim 1 , wherein the thermostable endopeptidase (i) has at least 60% sequence identity to the polypeptide of SEQ ID NO: 8, (ii) is encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5, or (iii) is a variant of the polypeptide of SEQ ID NO: 8 comprising a substitution, deletion, and/or insertion at one or more positions.

7. The method of claim 1 , wherein the thermostable endopeptidase (i) has at least 60% sequence identity to the polypeptide of SEQ ID NO: 4, or (ii) is a variant of the polypeptide of SEQ ID NO: 4 comprising a substitution, deletion, and/or insertion at one or more positions.

8. The method of claim 1 , wherein the protease preparation added in step c) is a protease preparation from Aspergillus.

9. The method of claim 1 , where in step d) the composition is incubated at a temperature of 30-65° C.

10. The method of claim 1 , wherein an enzyme capable of converting Gln to Glu is added at the same time or after step c).

11. The method of claim 1 , wherein the protein hydrolysate obtained in step d) has a degree of hydrolysis of at least 10%.

12. The method of claim 1 , wherein the substrate protein is selected from soy protein, wheat gluten protein or whey protein.

13. The method of claim 1 , wherein the composition comprising the substrate protein has a dry matter content of at least 1% (w/w).

14. The method of claim 1 , wherein the first hydrolysis step is performed at a temperature of 75-120° C. and the second hydrolysis step is performed at a temperature of 30-65° C.

15. The method of claim 1 , wherein the first hydrolysis step is performed at a temperature of 80-115° C. and the second hydrolysis step is performed at a temperature of 35-60° C.

16. The method of claim 1 , wherein the first hydrolysis step is performed at a temperature of 85-110° C. and the second hydrolysis step is performed at a temperature of 40-60° C.

17. The method of claim 1 , wherein the first hydrolysis step is performed at a temperature of 90-105° C. and the second hydrolysis step is performed at a temperature of 45-55° C.

18. The method of claim 1 , wherein the protein hydrolysate obtained by the method has a solubility of 60-100%.

19. The method of claim 1 , wherein the protein hydrolysate obtained by the method has a solubility of 65-100%.

20. The method of claim 1 , wherein the protein hydrolysate obtained by the method has a solubility of 70-100%.

Assignees

• Novozymes As

Inventors

• Lynglev Gitte Budolfsen

Classifications

• A23J3/346

  of vegetable proteins · CPC title

• A23J3/343

  of dairy proteins · CPC title

• C12Y304/11

  Aminopeptidases (3.4.11) · CPC title

• C12Y304/22

  Cysteine endopeptidases (3.4.22) · CPC title

• C12P21/06 Primary

  produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00) · CPC title