Method for producing a protein hydrolysate employing an aspergillus fumigatus tripeptidyl peptidase
AssigneeDupont Nutrition Biosciences Aps
InventorsKarsten Matthias Kragh, Svend Haaning, Ernst Meinjohanns, Peter Edvard Degn, Steffan Yde Bak, Thomas Eisele
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Abstract
The present invention relates to compositions and methods for the production of a hydrolysate comprising at least one endoprotease and a tripeptidyl peptidase capable of cleaving tripeptides from the N-terminus a peptide and/or proteins having one or more of lysine, arginine or glycine in the P1 position wherein said tripeptidyl peptidase is capable of being used at a temperature between 45° C. and 70° C.
Claims33
A method for the production of a hydrolysate comprising:
- a) admixing at least one protein or a portion thereof with a tripeptidyl peptidase which:
- i) comprises the amino acid sequence SEQ ID No. 3, SEQ ID No. 4 or a functional fragment thereof;
- ii) comprises an amino acid having at least 70% identity to SEQ ID No. 3 or SEQ ID No. 4;
- iii) is encoded by a nucleotide sequence comprising the sequence SEQ ID No. 1 or SEQ ID No. 2;
- iv) is encoded by a nucleotide sequence which has at least about 70% identity to SEQ ID No. 1 or SEQ ID No. 2;
- v) is encoded by a nucleotide sequence which hybridises to SEQ ID No. 1 or SEQ ID No. 2 under medium stringency conditions;
- or
- vi) is encoded by a nucleotide sequence which differs from SEQ ID No. 1 or SEQ ID No. 2 due to degeneracy of the genetic code;
- b) incubating at a temperature between 45° C. and 70° C., and
- c) recovering the hydrolysate.
A reaction system comprising at least one protein or a portion thereof and a tripeptidyl peptidase which:
- a) comprises the amino acid sequence SEQ ID No. 3, SEQ ID No. 4 or a functional fragment thereof;
- b) comprises an amino acid having at least 70% identity to SEQ ID No. 3 or SEQ ID No. 4;
- c) is encoded by a nucleotide sequence comprising the sequence SEQ ID No. 1 or SEQ ID No. 2;
- d) is encoded by a nucleotide sequence which has at least about 70% identity to SEQ ID No. 1 or SEQ ID No. 2;
- e) is encoded by a nucleotide sequence which hybridises to SEQ ID No. 1 or SEQ ID No. 2 under medium stringency conditions;
- or
- f) is encoded by a nucleotide sequence which differs from SEQ ID No. 1 or SEQ ID No. 2 due to degeneracy of the genetic code;
- wherein the reaction system is maintained at a temperature between 45° C. and 70° C. for a sufficient period of time to allow production of a hydrolysate.
A method for the expression of a tripeptidyl peptidase, wherein said method comprises:
- a) transforming a Trichderma host cell with a nucleic acid or vector comprising
- i) the nucleotide sequence SEQ ID No. 1 or SEQ ID No. 2;
- ii) a nucleotide sequence which has at least about 70% identity to SEQ ID No. 1 or SEQ ID No. 2;
- iii) a nucleotide sequence which hybridises to SEQ ID No. 1 or SEQ ID No. 2 under medium stringency conditions;
- or
- iv) a nucleotide sequence which differs from SEQ ID No. 1 or SEQ ID No. 2 due to degeneracy of the genetic code;
- b) expressing the nucleic acid sequence or vector of step a);
- and
- c) obtaining the tripeptidyl peptidase or a fermentate comprising said tripeptidyl peptidase and optionally isolating and/or purifying and/or packaging.
Use of a tripeptidyl peptidase which:
- a) comprises the amino acid sequence SEQ ID No. 3, SEQ ID No. 4 or a functional fragment thereof;
- b) comprises an amino acid having at least 70% identity to SEQ ID No. 3 or SEQ ID No. 4;
- c) is encoded by a nucleotide sequence comprising the sequence SEQ ID No. 1 or SEQ ID No. 2;
- d) is encoded by a nucleotide sequence which has at least about 70% identity to SEQ ID No. 1 or SEQ ID No. 2;
- e) is encoded by a nucleotide sequence which hybridises to SEQ ID No. 1 or SEQ ID No. 2 under medium stringency conditions;
- or
- f) is encoded by a nucleotide sequence which differs from SEQ ID No. 1 or SEQ ID No. 2 due to degeneracy of the genetic code;
- in the manufacture of a hydrolysate at a temperature between 45° C. and 70° C.
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Dependencies are inferred from the claim text.