Patent · US Expired

Process for bacterial production of polypeptides

US5789199A · kind A · utility

236Cited by
3References
32Claims
0Family size

Assignee

Inventors

Key dates

Filing dateOct 31, 1996
Grant dateAug 4, 1998
Priority date
Expiry dateOct 31, 2016

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC07K2319/00
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

A process is provided for producing a heterologous polypeptide in bacteria. This process comprises, in a first step, culturing bacterial cells that lack their native pstS gene and comprise nucleic acid encoding a PstS variant having an amino acid variation within the phosphate-binding region of the corresponding native PstS, nucleic acid encoding a DsbA or DsbC protein, nucleic acid encoding the heterologous polypeptide, a signal sequence for secretion of both the DsbA or DsbC protein and the heterologous polypeptide, an inducible promoter for the nucleic acid encoding the DsbA or DsbC protein, and an alkaline phosphatase promoter for the nucleic acid encoding the heterologous polypeptide. The nucleic acid encoding a PstS variant is under the transcriptional control of the wild-type pstS gene promoter. The second step of the process involves recovering the heterologous polypeptide from the periplasm or the culture medium.

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