Patent Number: 6,309,868

Title: Cloning of the prolyl-dipeptidyl-peptidase from Aspergillus oryzae

Abstract: 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.

Inventors: Monod; Michel (Lausanne, CH), Doumas; Agnes (Gollian, CH), Affolter; Michael (Pully, CH), Van Den Broek; Peter (Ch-Epalinges, CH)

Assignee: Nestec S.A.

International Classification: A23J 3/00 (20060101); A23J 3/30 (20060101); A23L 1/23 (20060101); A23L 1/226 (20060101); A23L 1/38 (20060101); A23L 1/36 (20060101); C12N 9/48 (20060101); C12N 009/14 (); C12N 001/20 (); C12N 001/14 (); C12N 015/00 (); C07H 021/04 ()

Expiration Date: 10/30/2018