Method for producing a protein hydrolysate using an endopeptidase and a carboxypeptidase

Patent metadata

Field	Value
Publication number	US-11946079-B2
Application number	US-202217571594-A
Country	US
Kind code	B2
Filing date	Jan 10, 2022
Priority date	Jun 9, 2017
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

The present invention relates to a method for producing a protein hydrolysate using a polypeptide having endopeptidase activity and a polypeptide having carboxypeptidase activity and the use of these enzymes for hydrolysing a protein substrate. In addition, the present invention relates to polypeptides having carboxypeptidase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

First claim

The invention claimed is:

1. A method for producing a protein hydrolysate comprising:
a. providing an aqueous solution or suspension of a protein substrate; and
b. exposing the aqueous solution or suspension of the protein substrate to a polypeptide having endopeptidase activity and to a polypeptide having carboxypeptidase activity, to obtain the protein hydrolysate;

wherein the polypeptide having carboxypeptidase activity comprises an amino acid sequence with at least 90% sequence identity to SEQ ID Nos: 2, 4, 6, 8, or 10 and also is characterised by having a Pro/ACHA*100 ratio of at least 30.

2. The method of claim 1 , wherein the concentration of the protein substrate in the aqueous solution or suspension is in the range of 5-35%.

3. The method of claim 1 , wherein the polypeptide having carboxypeptidase activity is selected from the group consisting of:
a. a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4 or SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10;
b. a polypeptide encoded by a polynucleotide having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, SEQ ID NO: 3 or SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9 or the cDNA sequence thereof; and
c. a variant of the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 or SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 positions.

Assignees

• Novozymes As

Inventors

• Hendriksen Hanne Vang
• Lynglev Gitte Budolfsen
• Frisner Henrik
• Liu Ciu
• Liu Ye
• Della Pia Eduardo Antonio
• Heldt-Hansen Hans Peter
• Jensen Kenneth
• Peng Wei
• Li Ming

Classifications

• C12N9/485 Primary

  Exopeptidases (3.4.11-3.4.19) · 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

• C12P13/005

  Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids · CPC title

• A23J3/34

  using enzymes · CPC title