Expression of mammalian proteins in <i>Pseudomonas fluorescens </i>

摘要

The invention is a process for improved production of a recombinant mammalian protein by expression in a Pseudomonad, particularly in a Pseudomonas fluorescens organism. The process improves production of mammalian proteins, particularly human or human-derived proteins, over known expression systems such as E. coli in comparable circumstances Processes for improved production of isolated mammalian, particularly human, proteins are provided.

主权项

1. A process for producing a recombinant mammalian protein in a Pseudomonas fluorescens host cell comprising:
a. transforming said Pseudomonas fluorescens host cell with a nucleic acid encoding the recombinant mammalian protein operably linked to a periplasmic secretion leader coding sequence; b. growing the transformed Pseudomonas fluorescens host cell under conditions that allow production of the recombinant mammalian protein, and secretion of the recombinant mammalian protein into the periplasm; and c. isolating the recombinant mammalian protein;wherein the recombinant mammalian protein is produced as 5-40% total cell protein, wherein the recombinant mammalian protein is present in the host cell in a soluble form, and wherein the soluble recombinant mammalian protein is produced at a titre in the P. fluorescens host cell that is about 1.5-fold to about 8-fold higher than the titre of the soluble recombinant mammalian protein produced in an E. coli host cell.