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 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.
Blackwell
J R
JR
Department of Biological Sciences, University of Wales, Aberystwyth, UK.
Horgan
R
R
D019883:Alkyl and Aryl Transferases; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012231:Rhizobium / Q000201:enzymology / Q000235:genetics; D014166:Transferases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D011994:Recombinant Proteins; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C024274:adenylate isopentenyltransferase