Patent ID: 7276605

Claim:
A method for the recombinant production of active trypsin comprising the steps: a) providing a yeast host cell comprising a recombinant nucleic acid that encodes a trypsinogen protein comprising a signal peptide and a shortened propeptide sequence that consists of no more than 10 amino acids, wherein the propeptide retains the amino acid sequence from position 20 to position 25 of SEQ ID NO:1 bound to the N-terminus of the amino acid sequence of trypsin, and wherein said signal peptide is bound to the N-terminus of said propeptide and mediates secretion of trypsinogen from the host cell, b) culturing the yeast host cell in a first culture medium under conditions promoting the growth of the host cell; c) culturing the host cell in a second culture medium having a pH of from 3.0 to 4.0 and inducing the expression of said recombinant nucleic acid whereby the yeast host cell expresses a trypsinogen comprising the shortened propeptide in the second culture medium significantly more than a trypsinogen comprising a complete propeptide; d) isolating the trypsinogen comprising the propeptide from the second culture medium, and e) cleaving the propeptide sequence by addition of trypsin and/or by autocatalytic cleavage to form active trypsin.