Patent · US Expired

Streptomyces proteases and methods for improved secretion of recombinantly-expressed proteins

US5856166A · kind A · utility

2Cited by
3References
20Claims
0Family size

Assignee

Inventors

Key dates

Filing dateJun 24, 1994
Grant dateJan 5, 1999
Priority date
Expiry dateJun 24, 2014

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC07K2319/02
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

A family of proteases endogenous to Streptomyces cells degrades heterologous proteins secreted from Streptomyces host cells. The previously unidentified proteases include (1) tripeptidyl aminopeptidase--Streptomyces ("Tap"), (2) a Streptomyces protease ("Ssp") which displayed significant amino acid sequence homology to Subtilisin BPN' and showed an ability to remove tripeptides from the amino termini of proteins and peptides, and (3) other proteases derived from Streptomyces which degraded certain substrates under certain conditions. Degradation was alleviated by selective inhibition of secreted proteases or by using hosts with impaired capabilities to produce proteases. An irreversible inhibitor was designed based upon the mechanism and substrate specificity of the target protease. Hosts secreting high amounts of proteases were selected. Impaired hosts were produced by deleting or altering the nucleotide sequence for the proteases.

Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.