Patent ID: 8759028
Filing Date: 2014-06-24
Classification: C07K,C12N,C12P,C12Y

Abstract:
1. A process for producing a target protein, comprising: providing a host cell having an outer membrane and harboring an expression cassette containing a recombinant polynucleotide therein, wherein the host cell is capable of expressing a fusion protein encoded by the recombinant polynucleotide and wherein the fusion protein comprises: (i) an anchoring protein that comprises a N-terminal amino acid sequence of an ice nucleation protein, so that the fusion protein, once expressed in the host cell, is directed by the anchoring protein to be anchored and exposed on the outer membrane of the host cell; (ii) the target protein; and (iii) a self-splicing protein that comprises a first end fused with the anchoring protein and a second end fused with the target protein, wherein the self-splicing protein comprises a N-terminal or C-terminal amino acid sequence of an intein protein at the second end thereof, such that upon an environmental stimulus, the self-splicing protein exerts a self-cleavage at the second end thereof to release the target protein from the fusion protein; obtaining a cell culture by culturing the host cell in a medium under a condition that enables the fusion protein to be expressed and anchored on the outer membrane of the host cell; subjecting the cell culture thus obtained to an environmental stimulus that induces the self-splicing protein to exert a self-cleavage at the second end thereof; and recovering the target protein by a separating treatment.