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AssigneesPresident and Fellows of Harvard College, Board of Regents, The University of Texas System
InventorsJonathan Beckwith, Fredrik Aslund, Paul H. Bessette, George Georgiou, Daniel Ritz, Jackie Eun-ah Lim
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The invention provides composition and methods for producing proteins of interest which comprise at least one disulfide bond, include proteins which in their mature form do not contain disulfide bonds, but whose precursor molecule contained at least one disulfide bond. The methods employ a host cell modified to more efficiently produce properly folded disulfide bond containing proteins. The host cells generally contain a mutation in one or more reductase genes, and can be further genetically modified to increase their growth rate, and are further optionally modified to increase the expression of a catalyst of disulfide bond formation. Host cells, methods for using such to produce proteins of interest, proteins of interest produced by these methods are within the scope of the invention.
A prokaryotic cell that is genetically modified to shift the redox status of the cytoplasm to a more oxidative state, which cell further expresses a heterologous polypeptide selected from the group consisting of catalysts of disulfide bond formation, disulfide bond isomerization and the combination of disulfide bond formation and disulfide bond isomerization, and which cell is further modified to increase its ability to proliferate, wherein the modification to increase its ability to proliferate is a mutation in an AhpC gene.
Dependencies are inferred from the claim text.