Expression of recombinant proteinase k <I> from tritirachium album </I>in yeast
AssigneeRoche Diagnostics Operations, Inc.
InventorsRainer Mueller, Johann-Peter Thalhofer, Frank Geipel, Stephan Glaser, Werner Hoelke, Helmut Schoen, Thomas Kirschbaum
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Abstract
The invention concerns a method for the expression of a gene coding for a soluble proteinase K in yeast e.g. in Pichia pastoris with subsequent secretion into the culture medium. In addition a method for purifying the heterologously expressed and secreted proteinase K is described.
Claims17
A method for producing recombinant proteinase K comprising the steps:
- a) transformating a host cell with a vector containing a DNA coding for the zymogenic precursor of proteinase K which is fused upstream of the coding sequence with a sequence in the reading frame which codes for a signal peptide and is under the control of a suitable promoter for the host cell,
- b) expressing of the zymogenic precursor of proteinase K
- c) secreting and autocatalytically activating proteinase K
- wherein the host cell is a yeast cell and the protein is secreted in a soluble form by this expression host.
A vector comprising DNA coding for the zymogenic precursor of proteinase K, wherein this DNA is fused upstream of the coding sequence with a sequence in the reading frame which codes for a suitable signal peptide, and the coding gene is under the control of a suitable promoter for the host cell and this vector is suitable for the transformation of yeast.
A vector as claimed in 10, wherein the vector is pPICPK-1 or pPICPK-2.
Dependencies are inferred from the claim text.