Expression of bovine lung prostaglandin F synthase in Escherichia coli

摘要: Summary The full-length bovine lung prostaglandin(PG) F synthase cDNA was constructed from partial clones and ligated into bacterial expression vector pUC8 to develop plasmid pUCPF1. This permitted the synthesis of PGF in Escherichia coli. recombinant bacteria overproduced a 36-KDa protein that recognized by anti-PGF antibody, expressed purified apparent homogeneity. reduced not only carbonyl compounds including PGD2and phenanthrenequinone but also PGH2; Km values for phenanthrenequinone, PGD2, PGH2of were 0.1, 100, 8μM, respectively, which are same as those synthase. produced PGF2αfrom PGH2, 9α,11β-PGF2from PGD2at different active sites. Moreover, structure Eschericia coli essentially identical native enzyme terms C-terminal sequence, sulfhydryl groups, CD spectra except nine amino acids provided lac Z′ gene fused N-terminus. These results indicate is We confirmed dual function catalyzing reduction PGH2and PGD2on single it has one binding site NADPH.