Expression of plastocyanin and cytochrome f of the cyanobacterium Phormidium laminosum in Escherichia coli and Paracoccus denitrificans and the role of leader peptides

The gene for plastocyanin from the cyanobacterium Phormidium laminosum was successfully expressed in Escherichia coli. Expression of the gene for cyto chrome f resulted in the production of holocytochrome f in the periplasmic space of E. coli, but the yield was low. Expression in Paracoccus dentrific ans yielded no holoprotein. When the region encoding the cytochrome f leade r sequence was replaced with more typical bacterial leader sequences (those from the P. laminosum plastocyanin gene and the Paracoccus I versutus cyto chrome c-550 gene), much higher yields were consistently obtained in both s pecies. Overexpressed proteins were compared to those isolated from P. lami nosum and found to be identical in mass, isoelectric point, redox midpoint potential and (for plastocyanin) H-1-NMR spectrum. (C) 1999 Published by El sevier Science B.V. All rights reserved.