Loading patent…
AssigneeNestec S.A.
InventorsMichel Monod, Agnes Doumas, Michael Affolter, Peter Van Den Broek
Estimating the term…
The invention has for object the new recombinant prolyl-dipeptidyl-peptidase enzyme (DPP IV) from Aspergillus oryzae comprising the amino-acid sequence from amino acid 1 to amino acid 755 of SEQ ID NO:2 or functional derivatives thereof, and providing a high level of hydrolyzing specificity towards proteins and peptides starting with X-Pro- thus liberating dipeptides of X-Pro type, wherein X is any amino acid. The invention also provides a DNA molecule encoding the enzyme according to the invention, cells expressing the enzyme according to the invention by recombinant technology, an Aspergillus naturally providing a prolyl-dipeptidyl-peptidase activity which has integrated multiple copies of the Aspergillus native promoter which naturally directs the expression of the gene encoding the prolyl-dipeptidyl-peptidase activity, Aspergillus naturally providing a prolyl-dipeptidyl-peptidase activity which is manipulated genetically so that the dppIV gene is inactivated. The invention provides a method for producing the enzyme according to the invention, comprising cultivating the cells of the invention in a suitable growth medium under conditions that the cells express the enzyme, and optionally isolating the enzyme in the form of a concentrate. The invention provides the use of the enzyme or the cells of the invention to hydrolyze protein containing materials. The invention provides the use of an enzyme and/or a cell providing a prolyl-dipeptidyl-peptidase activity, in combination with at least an enzyme providing a prolidase to hydrolyze protein containing materials. In a last further aspect, the invention provides a food product comprising a protein hydrolyzate obtainable by fermentation with at least a microorganism providing a prolyl-dipeptidyl-peptidase activity higher than 50 mU per ml when grown in a minimal medium containing 1% (w/v) of wheat gluten.
A recombinant prolyl-dipeptidyl-peptidase (DPP IV) from Aspergillus oryzae comprising the amino-acid sequence from amino acid 1 to amino acid 755 of SEQ ID NO:2 or polypeptides having at least 80% identity to amino acids 1-755 of SEQ ID NO:2 having prolyl-dipeptidyl peptidase activity.
A leader peptide of Aspergillus oryzae DPP IV having the amino-acid sequence from amino acid-16 to amino acid−1 of SEQ ID NO:2 or variants thereof having at least 80% identity to amino acids −16 to −1 of SEQ ID NO:2, wherein said variants are able to integrate the DPP IV into the cell wall or the cell membrane or secrete the enzyme into the periplasmic space or into the culture medium.
An Aspergillus which has integrated multiple copies of the promoter having the coding nucleotide sequence from nucleotide 1836 to nucleotide 1966 of SEQ ID NO:1.
An Aspergillus naturally providing a prolyl-dipeptidyl-peptidase activity which is manipulated genetically so that dppIV gene is inactivated.
Dependencies are inferred from the claim text.