Microbial protein expression system
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United States of America Patent
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Jul 19, 2005
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N/A
app pub date -
May 3, 2000
filing date -
May 4, 1999
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Expired
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Abstract
The present invention provides bacterial strains for secretion of soluble biologically active recombinant heterologous proteins into periplasm or on a surface/into a cultivation medium of bacteria. The invention exploits the secretion system of Gram-negative bacteria including periplasmic chaperones and usher/secretin proteins. For accomplishing the aim, the bacterial strains simultaneously express the fusion protein (signal peptide)-(mature hecterologous protein)-(subunit of a bacterial surface structure, Caf1), periplasmic chaperone specific for the subunit, and outer membrane usher/secretin protein specific for the subunit. Secretion of fusion proteins: (signal peptide of Caf1)-(mature human IL-1β)-(mature Caf1), (signal peptide of Caf1)-(mature human GM-CSF)-(mature Caf1), and (signal peptide of Caf1)-(mature human IL-1ra)-(mature Caf1) that were expressed in Escherichia coli simultaneously with the periplasmic chaperone Caf1M and the usher/secretin protein Caf1A are examples of the use of the invention.
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