J. Hill et al., HIGH-LEVEL EXPRESSION AND CHARACTERIZATION OF PLASMEPSIN II, AN ASPARTIC PROTEINASE FROM PLASMODIUM-FALCIPARUM, FEBS letters, 352(2), 1994, pp. 155-158
DNA encoding the last 48 residues of the propart and the whole mature
sequence of Plasmepsin II was inserted into the T7 dependent vector pE
T 3a for expression in E. coli. The resultant product was insoluble bu
t accumulated at similar to 20 mg/l of cell culture. Following solubil
isation with urea, the zymogen was refolded and, after purification by
ion-exchange chromatography, was autoactivated to generate mature Pla
smepsin II. The ability of this enzyme to hydrolyse several chromogeni
c peptide substrates was examined; despite an overall identity of simi
lar to 35% to human renin, Plasmepsin II was not inhibited significant
ly by renin inhibitors.