PROKARYOTIC HOST CELLS FOR EXPRESSING PROTEINS RICH IN DISULFIDE BONDS

摘要

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 prokaryotic 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 are further genetically modified to increase their growth rate by a mutation in an ahpC gene of the prokaryotic cell or the introduction into the prokaryotic cell of a nucleic acid encoding a mutated ahpC protein or portion thereof comprising a mutation, wherein the mutation is located around the amino acid region that is located between Cys46 and Cysl65 of the protein AhpC encoded by the ahpC gene or portion thereof. The host cells 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.