NOVEL METHODS, POLYPEPTIDES AND USES THEREOF

摘要

The present invention provides methods for the production of recombinant polypeptides having serine protease activity, polypeptides obtainable by such methods and use of said polypeptides in medicine, cosmetics and industry. In particular, the invention provides recombinantly expressed mutants of trypsin I from Atlantic cod, which mutants exhibit improved stability and/or catalytic properties relative to the wildtype trypsin purified from cod.

主权项

1. A method for the production of a recombinant polypeptide having serine protease activity comprising
(a) transforming a microbial host cell, or population thereof, with a nucleic acid molecule encoding a zymogen polypeptide comprising an activation peptide fused to the N-terminus of a polypeptide having serine protease activity
wherein the zymogen polypeptide lacks a signal sequence; (b) expressing said zymogen polypeptide in the host cell(s) as inclusion bodies; (c) purifying the zymogen polypeptide from the host cell(s); and (d) activating the zymogen polypeptide by exposure to a protease, such as a trypsinwherein step (c) comprises solubilising the zymogen polypeptide from the inclusion bodies and refolding the polypeptide into a bioactive form.