Nestec S.a.

Societe Des Produits Nestle S.a.

Peter Van Den Broek

Michael Affolter

Elizabeth Offord Cavin

Karine Vidal

Anne Donnet-Hughes

Michel Monod

Johannes De Reu

Elizabeth Offord-Cavin

Agnes Doumas

The present invention pertains to osteoprotegerin obtainable from milk sources, in particular human and bovine milk. The present invention also relates to the use thereof for preparing an ingestible preparation and/or a pharmaceutical composition, in particular to the use of such a preparation/composition for preventing or treating disorders associated with bone metabolism and immune function.

Osteoprotegerin in milk

The present invention refers to a koji mold capable of expressing proteolytic enzymes in the presence of a carbon source in at least the same amount as in the absence thereof. In particular, the present invention pertains to a mutation in the creA gene as a tool to increase the amount of a wide spectrum of proteolytic enzymes in the presence of a carbon source.

Expression of proteolytic enzymes in koji mold in the presence of carbon sources

The invention has for object the new recombinant prolyl-dipeptidyl-peptidase enzyme (DPP IV) from,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.

Cloning of the prolyl-dipeptidyl-peptidase from Aspergillus oryzae

Enhanced expression of proteolytic enzymes in koji mold

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WJ Maarn, Netherlands

Peter Van Den Broek