Method of improving taste and/or flavour of foods and beverages
The present invention discloses a method for producing foods and/or beverage having improved taste and/or flavour, comprising reacting a microbial aminopeptidase on a protein material optionally under the co-existence of a protease, wherein said aminopeptidase has the properties of: (a) having an activity of catalyzing the reaction of specifically releasing a glutamic acid and an aspartic acid from the N-terminal of a peptide and/or a protein; (b) having 50% or more activity at pH6.0-9.0 as compared with the activity at the optimum pH; (c) having 40% or more activity after heating at 25-60° C., pH7.5 for 30 minutes as compared with the activity of the non-heated enzyme; (d) having a molecular weight of about 40-60 kD as measured by SDS-PAGE and about 300-480 kD as measured by native-PAGE; (e) having a hydrolyzing activity of the aminopeptidase toward Glu-Glu dipeptide is 5 U/mg or more, preferably 10 U/mg or more.
1. A method for producing a food and/or beverage comprising reacting a protein in the food and/or beverage with an aminopeptidase wherein the aminopeptidase is encoded by the nucleic acid sequence of SEQ ID NO:6 or by a polynucleotide homolog that varies from SEQ ID NO: 6 as a result of degeneracy in the genetic code, wherein said polynucleotide homolog encodes the same amino acid sequence as the polynucleotide sequence of SEQ ID NO:6.
2. The method according to claim 1 , wherein the aminopeptidase is encoded the nucleotide sequence shown in SEQ ID NO:6.
3. The method according to claim 1 , wherein the aminopeptidase is produced by a microorganism transformed with polynucleotide having the sequence shown in SEQ ID NO:6.
4. The method according to claim 1 , wherein the foods and/or beverages are selected from the group consisting of protein hydrolysates, cheeses, tomato juice containing beverages and soy milk-containing beverages.
5. The method according to claim 1 , wherein reacting said protein in a food and/or beverage with an aminopeptidase further comprises the presence of a protease.
6. The method according to claim 5 , wherein said aminopeptidase is encoded by said polynucleotide homolog.
7. The method according to claim 5 , wherein said protease is a proteolytic enzyme produced by Aspergillus or Bacillus.
8. The method according to claim 5 , wherein said protease is selected from the group of enzyme preparations consisting of UMAMIZYME™, PROTEASE M™, FLAVOURZYME™ and ALCALASE™.
9. A method for producing a food and/or beverage comprising reacting a protein in the food and/or beverage with an aminopeptidase, wherein the aminopeptidase has the amino acid sequence of SEQ ID NO:7 or a homolog thereof, wherein said homolog thereof has an amino acid sequence of SEQ ID NO:7 having from one to ten amino acid replacements, deletions or additions, or combinations thereof and wherein said aminopeptidase homolog:
(a) has an activity of catalyzing the reaction of specifically releasing a glutamic acid or an aspartic acid from the N-terminal of a peptide and/or a protein;
(b) has 50% or more of the activity defined in clause (a) at pH 6.0-9.0 as compared with the activity at pH 7.5;
(c) has 40% or more of the activity defined in clause (a) after heating at 25-60° C., pH 7.5 for 30 minutes as compared with the activity of the aminopeptidase prior to heating;
(d) has a molecular weight of about 40-60 kD as measured by SOS-PAGE and about 300-480 kD as measured by native-PAGE; and
(e) has a hydrolyzing activity of 5 U/mg or more toward Glu-Glu dipeptide, wherein one U (unit) is an enzyme activity that liberates 1 micromole of Glu per 1 minute at 37° C.
10. The method according to claim 9 , wherein the aminopeptidase has the amino acid sequence of SEQ ID NO: 7.
11. The method according to claim 9 , wherein the aminopeptidase is said homolog thereof.
12. The method according to claim 9 , wherein the foods and/or beverages are selected from the group consisting of protein hydrolysates, cheeses, tomato juice containing beverages and soy milk-containing beverages.
13. The method according to claim 9 , wherein reacting said protein in a food and/or beverage with an aminopeptidase further comprises the presence of a protease.
14. The method according to claim 13 , wherein said protease is a proteolytic enzyme produced by Aspergillus or Bacillus.
15. The method according to claim 14 , wherein said protease is selected from the group of enzyme preparations consisting of UMAMIZYME™, PROTEASE M™, FLAVOURZYME™ and ALCALASE™.