Method for the prevention or reduction of haze in beverages
View Patent ↗The invention relates to a method for the prevention or reduction of haze in a beverage by the addition of an prolyl-specific endoprotease and to new beverages obtainable by the method according to the invention. It also relates to new endoproteases. Sequence information of a genomic DNA, cDNA as well as protein sequences.
1. A proteolytic method to reduce haze in a beverage, which method comprises adding a proline-specific endoprotease protein-containing preparation to the beverage, wherein the proline-specific endoprotease cuts a protein or peptide at places where the protein or peptide contains a prolyl residue to proteolytically reduce haze in the beverage, wherein at least 70% of the proteins in the preparation are said proline-specific endoprotease, the proline-specific endoprotease being comprised of an amino acid sequence which has at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 5.
2. The method of claim 1 , wherein said endoprotease has maximum prolyl-specific activity at a pH corresponding to the pH of the beverage to which it is added.
3. The method of claim 1 , wherein the beverage contains at least one protein.
4. The method of claim 3 , wherein the beverage contains polyphenols.
5. The method of claim 1 , wherein the beverage has a pH value below 7.
6. The method of claim 3 , wherein at least 150 milli-units of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
7. The method of claim 6 , wherein at least 500 milli-units of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
8. The method of claim 7 , wherein at least 1 unit of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
9. The method of claim 1 , wherein the beverage is beer.
10. The method of claim 1 , wherein the beverage is wine.
11. The method of claim 1 , wherein the beverage is fruit juice.
12. The method of claim 9 , wherein the proline-specific endoprotease protein containing preparation is added to a mash.
13. The method of claim 9 , wherein the proline-specific endoprotease protein containing preparation is added to a beer before haze is formed.
14. The method of claim 9 , wherein the proline-specific endoprotease protein containing preparation is added to a fermented beer after haze has been formed.
15. The method of claim 10 , wherein the proline-specific endoprotease protein containing preparation is added to a fermented wine.
16. The method of claim 1 , wherein the proline-specific endoprotease is isolated or purified prior to addition to the beverage.
17. A method to reduce haze in a beverage, which method comprises adding a proline-specific endoprotease to the beverage to reduce haze therein, wherein the proline-specific endoprotease cuts a protein or peptide at places where the protein or peptide contains a prolyl residue, wherein the proline-specific endoprotease is comprised of an amino acid sequence which has at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 5, wherein the proline-specific endoprotease is produced by recombinant expression, and wherein the proline-specific endoprotease is added as a filtrate obtained from a fermentation broth of cells which are modified to contain a recombinant nucleic acid construct for production of the proline-specific endoprotease, and wherein said filtrate contains proteins and at least 70% of the proteins in the filtrate are said proline-specific endoprotease.
18. A method to reduce haze in a beverage, which method comprises adding a proline-specific endoprotease to the beverage to reduce haze therein, wherein the proline-specific endoprotease is encoded by a polynucleotide which hybridizes under high stringency conditions of 0.3M sodium chloride, 0.03M sodium citrate, and 60° C. with the complement of the nucleotide sequence of SEQ ID NO: 1, and wherein the proline-specific endoprotease is produced by recombinant expression said proline-specific endoprotease being added to the beverage in a protein containing preparation wherein at least 70% of the proteins in the preparation are said proline-specific endoprotease.
19. A proteolytic method to reduce haze in a beverage, which method comprises adding a proline-specific endoprotease protein-containing preparation to the beverage to proteolytically reduce haze therein, wherein at least 70% of the proteins in the preparation are said proline-specific endoproteinase, the proline-specific endoprotease being comprised of the amino acid sequence of SEQ ID NO: 5.
20. The method of claim 18 , wherein the proline-specific endoprotease is encoded by a polynucleotide comprised of the nucleotide sequence of SEQ ID NO: 1.
21. The method of claim 18 , wherein the beverage contains at least one protein.
22. The method of claim 21 , wherein the beverage contains polyphenols.
23. The method of claim 18 , wherein the beverage has a pH value below 7.
24. The method of claim 21 , wherein at least 150 milli-units of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
25. The method of claim 24 , wherein at least 500 milli-units of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
26. The method of claim 25 , wherein at least 1 unit of proline-specific endoprotease activity, which can be determined using Z-Gly-Pro-pNA as substrate, is added to the beverage per gram protein in the beverage.
27. The method of claim 18 , wherein the beverage is beer.
28. The method of claim 18 , wherein the beverage is wine.
29. The method of claim 18 , wherein the beverage is fruit juice.
30. The method of claim 27 , wherein the proline-specific endoprotease protein containing preparation is added to a mash.
31. The method of claim 27 , wherein the proline-specific endoprotease protein containing preparation is added to a beer before haze is formed.
32. The method of claim 27 , wherein the proline-specific endoprotease protein containing preparation is added to a fermented beer after haze has been formed.
33. The method of claim 28 , wherein the proline-specific endoprotease protein containing preparation is added to a fermented wine.
34. The method of claim 18 , wherein the proline-specific endoprotease is isolated or purified prior to addition to the beverage.
35. A method to reduce haze in a beverage, which method comprises adding a proline-specific endoprotease to the beverage to reduce haze therein, wherein the proline-specific endoprotease is comprised of the amino acid sequence of SEQ ID NO: 5, wherein the proline-specific endoprotease is produced by recombinant expression, and wherein the proline-specific endoprotease is added as a filtrate obtained from a fermentation broth of cells which are modified to contain a recombinant nucleic acid construct for production of the proline-specific endoprotease, and wherein said filtrate contains proteins and at least 70% of the proteins in the filtrate are said proline-specific endoprotease.