Method for producing therapeutic proteins in <i>Pichia pastoris </i>lacking dipeptidyl aminopeptidase activity

摘要

The present invention related to methods and compositions for producing therapeutic proteins in yeast cell lines, and in particular Pichia pastoris, lacking dipeptidyl aminopeptidase (DAP) activity. DAP activity has been eliminated by genetically modifying a Pichia pastoris cell line such that STE13 and DAP2 have been deleted.

主权项

1. A method for producing a recombinant protein having an X-P/A at the amino-terminus, wherein X is any amino acid and P/A is proline or alanine, in a Pichia host cell, wherein the recombinant protein is secreted into the host cell culture medium, in which STE13 and DAP2 has been deleted or disrupted, comprising:
a. transforming the Pichia cell with a polynucleotide vector encoding the protein having the X-P/A at the amino-terminus; b. culturing the transformed Pichia cell in a medium under conditions which induce expression of the protein; and c. isolating the protein from the culture medium.