[Research on renaturation of recombinant human pro-urokinase expressed from Escherichia coli].
Recombinant human pro-urokinase forms insoluble inclusion body when overexpressed in Escherichia coli, and it must be denatured and renatured in vitro before it acquires activity. This study aimed to increase the renaturation yield of denatured pro-urokinase. We have evaluated the basic renaturation conditions of pro-urokinase through qualitative and quantitative analysis of pH, temperature, denaturant concentration, protein concentration, the ratio of reduced and oxidized thiol reagents. The effects of nonspecific additives, step-wise dilution and urea gradient dialysis have been also compared. The optimal conditions of pro-urokinase renaturation with the yield about 20%-30% have been obtained.
Zhu
H
H
Department of Biochemistry, Nanjing University.
Liu
W
W
Shi
W
W
Xue
Y M
YM
Ma
Z
Z
D004229:Dithiothreitol / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D014568:Urokinase-Type Plasminogen Activator / Q000737:chemistry*
D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator; D004229:Dithiothreitol; C067551:saruplase