PMID:21824411
| Citation |
Šiurkus, J and Neubauer, P (2011) Heterologous production of active ribonuclease inhibitor in Escherichia coli by redox state control and chaperonin coexpression. Microb. Cell Fact. 10:65 |
|---|---|
| Abstract |
Eukaryotic Ribonuclease inhibitor (RI), belonging to the RNH1 family, is distinguished by unique features--a high sensitivity to oxidation due to the large number of reduced cysteins and a high hydrophobicity, which made most production approaches so far unsuccessful or resulted in very low yields. In this work efficient in vivo folding of native RI in the Escherichia coli cytoplasm was obtained by external addition of a reducing agent in tandem with oxygen limitation and overproduction of a molecular chaperonin. After optimisation of the production conditions in the shake flask scale the process was scaled up to high cell densities by applying a glucose limited fed-batch procedure. |
| Links |
PubMed PMC3161860 Online version:10.1186/1475-2859-10-65 |
| Keywords |
Bioreactors; Chaperonin 60/genetics; Chaperonin 60/metabolism; Cloning, Molecular; Enzyme Inhibitors/metabolism; Escherichia coli/metabolism; Oxidation-Reduction; Oxygen/chemistry; Oxygen/pharmacology; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Reducing Agents/chemistry; Reducing Agents/pharmacology; Ribonucleases/antagonists & inhibitors; Temperature |
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