Purified human growth hormone from E. coli is biologically active

The purification of eukaryotic polypeptides synthesized in Escherichia coli

“…Receptor binding of human complement fragment C5a (Mandecki et al, 1985) and biological activity of FMDV proteinase (Klump et al, 1984) were demonstrated using unpurified lysis supernatants. Pure retroviral p24 gag protein was purified from E. coli in a single immunopurification step (Dowbenko et al, 1985) while a five-step process was used to produce human growth hormone (Olsen et al, 1981). Other examples of soluble proteins are human IFN-a (Staehelin et al, 1981;De Maeyer et al, 1982) boVine IFN-a (Grosfeld et al, 1985), chicken triosephosphate isomerase (Straus & Gilbert, 1985), human lymphotoxin (Gray et al, 1984), human TNF (Pennica et al, 1984) and murine TNF .…”

“…E. coli does possess an aminopeptidase with broad substrate specificity, but the efficiency of methionine removal from recombinant polypeptides is variable. The N-terminal methionine residues were completely removed from IFN-, (Stebbing et al, 1982) and bovine growth hormone (George et al, 1985), whilst there was differential processing of IFN-a (Staehelin et al, 1981) and human growth hormone (Olsen et al, 1981). With an N-terminal sequence of Met-Ala-Pro-, IL-2 was completely unprocessed (Liang et al, 1985).…”

“…The advantage of using fusion proteins or secretion is that processing yields the correct N-terminal residue. It is possible for the presence of an N-terminal methionine to have no effect on biological activity, as demonstrated for human growth hormone and IL-2 (Olsen et al, 1981;Liang et al, 1985).…”

High purity recombinant human growth hormone (rhGH) expression in Escherichia coli under phoA promoter

“…5 In the past years, significant progress has been made in recombinant hGH (rhGH) production. 6 Zamani et al optimized the cultivation medium of recombinant E. coli through response surface methodology, giving hGH yield of 391 mg/L. 7 However, hGH was expressed as inclusion body in E. coli, which was a major hurdle.…”

The construction of a syntheticEscherichia coli trppromoter and its use in the expression of a synthetic interferon gene

“…In the latter case either a normal eukaryotic protein or one carrying an additional N-terminal methionine residue is produced (10,11). Unless a hybrid polypeptide product has some advantageous property, e.g.…”