Synthesis of Rhodobacter sphaeroides cytochrome c2 in Escherichia coli

Abstract

The cytochrome c2 structural gene, cycA, from Rhodobacter sphaeroides was expressed in Escherichia coli. CycA-specific mRNA was detected in E. coli both under aerobic and anaerobic conditions with trimethylamine-N-oxide as electron acceptor. However mature holocytochrome c2 was only detected in anaerobically-grown cells. The mature form of cytochrome c2 (Mr = 12,500) was secreted into the periplasm of E. coli suggesting that the signal polypeptide was processed. The cytochrome c2 synthesized in E. coli exhibited absorbance maxima in the reduced form at 550 nm (alpha-band) and 521 nm (beta-band) and contained covalently attached haem c. The results indicate that a foreign c-type cytochrome can be secreted and assembled in E. coli under anaerobic conditions.