A novel strategy for production of a highly expressed recombinant protein in an active form

Under standard growth conditions, E. coli transformed with the high-level expression vector pMON5525 produces recombinant DMAPP/AMP transferase in inactive, insoluble complexes. We have produced large amounts of active, soluble protein by growing and inducing the cells under osmotic stress in the pr...

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Bibliographic Details
Published in:FEBS letters 1991-12, Vol.295 (1), p.10-12
Main Authors: Blackwell, John R., Horgan, Roger
Format: Article
Language:English
Subjects:
Activity
Alkyl and Aryl Transferases
Biological and medical sciences
Biotechnology
DMAPP/AMP transferase
E. coli
Electrophoresis, Polyacrylamide Gel
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Kinetics
Methods. Procedures. Technologies
Protein engineering
Recombinant protein
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
Rhizobium - enzymology
Rhizobium - genetics
Transferases - biosynthesis
Transferases - genetics
Transferases - isolation & purification
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Description
Summary:Under standard growth conditions, E. coli transformed with the high-level expression vector pMON5525 produces recombinant DMAPP/AMP transferase in inactive, insoluble complexes. We have produced large amounts of active, soluble protein by growing and inducing the cells under osmotic stress in the presence of sorbitol and glycyl betaine. This caused an increase of up to 427-fold in the active yield, and the disappearance of the protein from the pelletable fraction of cell extracts. This treatment may have wide applicability.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(91)81372-F