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EP-4739120-A1 - FOAMABLE DAIRY PRODUCTS

EP4739120A1EP 4739120 A1EP4739120 A1EP 4739120A1EP-4739120-A1

Abstract

The invention provides a dairy composition with improved aeration properties, where the dairy composition comprises a protein deamidase and optionally a second polypeptide derived from the propeptide of a protein deamidase proform. The invention further provides an improved method for producing aerated dairy products from dairy compositions, where a protein deamidase and optionally a second polypeptide derived from the propeptide of a protein deamidase proform- has been added to the dairy composition.

Inventors

  • KALUM, LISBETH

Assignees

  • Novozymes A/S

Dates

Publication Date
20260513
Application Date
20240628

Claims (15)

  1. 1. A method for producing improved aerated dairy products, comprising: a) obtaining a dairy composition; b) adding a first polypeptide to the dairy composition, wherein the first polypeptide is a mature protein deamidase derived from a proform protein deamidase; c) optionally adding a second polypeptide to the dairy composition, wherein the second polypeptide is a propeptide derived from the same proform protein deamidase; and d) aerating the dairy composition to produce an aerated dairy product; wherein the aerated dairy product is improved compared to a similar method where the first polypeptide and optionally the second polypeptide is not added to the dairy composition.
  2. 2. The method of claim 1 , wherein the dairy composition comprises cream, milk, butter, concentrated milk, condensed milk, evaporated milk, reconstituted milk, fermented milk, milk protein concentrate, whey protein concentrate, or a combination thereof.
  3. 3. The method of claim 1 or 2, wherein the mature protein deamidase comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NOs: 2, 5, 8, or 11.
  4. 4. The method any one of the proceeding claims, wherein the second polypeptide is a propeptide which comprises a mutation which decreases its binding affinity for the deamidase domain compared to the parent propeptide.
  5. 5. The method of any one of the proceeding claims, wherein the second polypeptide is a propeptide which comprises: a) an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 3 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 3; b) an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 6 and has an alanine at the position corresponding to position 99 of SEQ ID NO: 6; c) an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 9 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 9; or d) an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 12 and has a glycine at the position corresponding to position 101 of SEQ ID NO: 12.
  6. 6. The method of any one of the proceeding claims, wherein the second polypeptide comprises the amino acid sequence of SEQ ID NOs: 16, 17, 18, or 19.
  7. 7. The method of any one of the proceeding claims, wherein: a) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 3 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 3; b) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 5, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 6 and has an alanine at the position corresponding to position 99 of SEQ ID NO: 6; c) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 8, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 9 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 9; or d) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 11 , and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 12 and has a glycine at the position corresponding to position 101 of SEQ ID NO:
  8. 8. A dairy composition comprising a first polypeptide, wherein the first polypeptide is a mature protein deamidase derived from a proform protein deamidase, and optionally a second polypeptide, wherein the second polypeptide is a propeptide derived from the same proform protein deamidase.
  9. 9. The dairy composition of claim 8, wherein: a) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 3 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 3; b) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 5, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 6 and has an alanine at the position corresponding to position 99 of SEQ ID NO: 6; c) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 8, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 9 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 9; or d) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 11 , and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 12 and has a glycine at the position corresponding to position 101 of SEQ ID NO: 12.
  10. 10. The dairy composition of claim 8 or 9, wherein the dairy composition comprises cream, milk, butter, concentrated milk, evaporated milk, condensed milk, reconstituted milk, fermented milk, milk protein concentrate, whey protein concentrate, or a combination thereof.
  11. 11. The dairy composition of claims 8 to 10, wherein the dairy composition is processed to produce aerated dairy products, such as ice cream, whipped cream, whipped yoghurt, whipped cream cheese, foamed milk, or whipped butter.
  12. 12. An aerated dairy product produced by the method of any of claims 1 to 7.
  13. 13. The aerated dairy product of claim 12, wherein the aerated dairy product is ice cream, whipped cream, whipped yoghurt, whipped cream cheese, foamed milk, whipped butter, or whipped butter spread.
  14. 14. Use of a first polypeptide, wherein the first polypeptide is a mature protein deamidase derived from a proform protein deamidase, and optionally a second polypeptide, wherein the second polypeptide is a propeptide derived from the same proform protein deamidase, to produce a dairy composition with improved aeration or foamability.
  15. 15. The use of claim 14, wherein: a) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 3 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 3; b) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 5, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 6 and has an alanine at the position corresponding to position 99 of SEQ ID NO: 6; c) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 8, and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 9 and has a glycine at the position corresponding to position 99 of SEQ ID NO: 9; or d) the first polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 11 , and the optional second polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 12 and has a glycine at the position corresponding to position 101 of SEQ ID NO: 12.

Description

FOAMABLE DAIRY PRODUCTS Reference to sequence listing This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference. FIELD OF THE INVENTION The present invention relates to dairy products that are treated to enhance the foamability of the dairy product. The present invention also provides a method of treating the dairy product to enhance foamability. BACKGROUND OF THE INVENTION Cappuccino-style beverages, which typically feature a mix of coffee and milk and a head of milk foam on top, frequently with a design, or “latte art”, in the top layer of the foam, are very popular today. Milk provides both a creamy texture to the beverage and a smoothness to the foam. Typically, whole milk is used, and frequently preferred, for the beverage. However, many health-conscious consumers prefer a lower fat milk, particularly milks that contain less than 2% milkfat, or even skim milk, which contains a very minimal amount of milkfat. Unfortunately, it is difficult to produce a foam of the same quality using low fat milk or skim milk compared to a foam produced using whole milk. Additionally, sometimes even whole milk can be difficult to foam to a desirable consistency and texture. SUMMARY OF THE INVENTION The invention provides a dairy composition with improved aeration properties, where the dairy composition comprises a first polypeptide which is a mature protein deamidase and optionally a second polypeptide which is a propeptide derived from a protein deamidase proform. The invention further provides an improved method for producing aerated dairy products from dairy compositions, where a protein deamidase and optionally a second polypeptide derived from the propeptide of the protein deamidase proform has been added to the dairy composition. Unexpectedly, the presence of the propeptide in the dairy composition improves the foamability of the dairy composition, producing an improved aerated dairy product compared to a method where only the mature protein deamidase is added. The invention also provides a method for improved foaming of a dairy composition, where the dairy composition comprises a first polypeptide which is a mature protein deamidase and optionally a second polypeptide which is a propeptide derived from a protein deamidase proform. The invention further provides a method for improving latte art produced from a dairy composition, where the dairy composition comprises a first polypeptide which is a mature protein deamidase and optionally a second polypeptide which is a propeptide derived from a protein deamidase proform. SEQUENCES SEQ ID NO: 1 : Amino acid sequence of proform polypeptide of deamidase from Chryseobacterium viscerum comprising the propeptide variant of F99G. SEQ ID NO: 2: Amino acid sequence of mature deamidase derived from SEQ ID NO: 1. SEQ ID NO: 3: Amino acid sequence of propeptide from Chryseobacterium viscerum comprising F99 (wild-type). SEQ ID NO: 4: Amino acid sequence of proform polypeptide of deamidase from Chryseobacterium gambrini. SEQ ID NO: 5: Amino acid sequence of mature deamidase derived from SEQ ID NO: 4. SEQ ID NO: 6: Amino acid sequence of propeptide derived from SEQ ID NO: 4. SEQ ID NO: 7: Amino acid sequence of proform polypeptide of deamidase from Chryseobacterium culicis. SEQ ID NO: 8: Amino acid sequence of mature deamidase derived from SEQ ID NO: 7. SEQ ID NO: 9: Amino acid sequence of propeptide derived from SEQ ID NO: 7. SEQ ID NO: 10: Amino acid sequence of proform polypeptide of deamidase from Chryseobacterium defluvii. SEQ ID NO: 11 : Amino acid sequence of mature deamidase derived from SEQ ID NO: 10. SEQ ID NO: 12: Amino acid sequence of propeptide derived from SEQ ID NO: 10. SEQ ID NO: 13: Amino acid sequence of proform polypeptide of deamidase from Chryseobacterium proteolyticum. SEQ ID NO: 14: Amino acid sequence of mature deamidase derived from SEQ ID NO: 13. SEQ ID NO: 15: Amino acid sequence of propeptide derived from SEQ ID NO: 13. SEQ ID NO: 16: Propeptide variant F99G of SEQ ID NO: 3. SEQ ID NO: 17: Propeptide variant F99A of SEQ ID NO: 6. SEQ ID NO: 18: Propeptide variant F99G of SEQ ID NO: 9. SEQ ID NO: 19: Propeptide variant L101G of SEQ ID NO: 12. SEQ ID NO: 20: Propeptide variant L100G of SEQ ID NO: 15. SEQ ID NO 21 : Variant of SEQ ID NO: 4 comprising the propeptide variant of SEQ ID NO: 17. SEQ ID NO 22: Variant of SEQ ID NO: 7 comprising the propeptide variant of SEQ ID NO: 18. SEQ ID NO 23: Variant of SEQ ID NO: 10 comprising the propeptide variant of SEQ ID NO: 19. SEQ ID NO 24: Variant of SEQ ID NO: 13 comprising the propeptide variant of SEQ ID NO: 20. DEFINITIONS Deamidase: The term “deamidase” means a protein-glutamine glutaminase (also known as glu- taminylpeptide glutaminase) activity, as described in EC 3.5.1.44, which catalyzes the hydrolysis of the gamma-amide of glutamine substituted at the carboxyl position or both the alpha-amino and carboxyl positions, e.g., L-