Patent ID: 6258560
Filing Date: 2001-07-10
Classification: C07K,C12N,C12P

Abstract:
A process for recovering a heterologous polypeptide from bacterial cells comprising:(a) culturing the bacterial cells which comprise a first nucleic acid encoding phage lysozyme and the gene t product, and a second nucleic acid encoding a protein that displays DNA-digesting activity under control of a signal sequence for secretion of the DNA-digesting protein, wherein the first and second nucleic acids are operatively linked to a first promoter that is the same for both and wherein both are linked on the same nucleic acid construct, and a third nucleic acid encoding the heterologous polypeptide, which third nucleic acid is linked to a second promoter, wherein the second promoter is inducible and the first promoter is different from the second promoter and is either (i) inducible or (ii) a weak constitutive promoter that does not require the addition of an inducer to function as a promoter,(b) adding an inducer specific for induction of expression of the nucleic acid encoding the heterologous polypeptide from the second inducible promoter,(c) optionally, when the first promoter driving expression of the nucleic acids encoding the phage lysozyme, the gene t product, and DNA-digesting protein is an inducible promoter, adding an inducer specific for the first promoter after accumulation of about 50% or more of the maximum accumulation of the heterologous polypeptide to be recovered,(d) lysing the cells, and(e) recovering accumulated heterologous polypeptide from the broth lysate.