AN EFFICIENT SYSTEM FOR PRODUCTION OF RECOMBINANT UROKINASE-TYPE PLASMINOGEN-ACTIVATOR

A system was developed to produce recombinant urokinase-type plasminog en activator in Escherichia coli. The urokinase-type plasminogen activ ator was produced with a 6x His-tag at the C-terminus which was shown to have the same activity, after refolding, as the wild-type protein, Purification of the recombinant protein to homogeneity (95%) was possi ble by one-step affinity chromatography under denaturing conditions. A s a result, proteolysis by bacterial proteases during purification was avoided. A higher refolding efficiency (40%) and a higher total recov ery yield (25%) of the recombinant protein were obtained by this metho d. (C) 1997 Academic Press.