Synthesis of the Rhodopseudomonas viridis holo-cytochrome c2 in Paracoccus denitrificans |
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| Authors: | Ernst Gerhus Reinhard Grisshammer Hartmut Michel Bernd Ludwig reas Turba |
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| Affiliation: | Institute of Biochemistry, Medical University, Lübeck, FRG; Centre for Protein Engineering, MRC, Cambridge, UK; Max Planck Institute for Biophysics, Frankfurt, FRG; Institute of Biochemistry, Molecular Genetics Group, Johann Wolfgang Goethe Universität, Robert-Mayer-Str. 7-9, D-60054 Frankfurt, FRG |
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| Abstract: | Abstract The gene encoding the Rhodopseudomonas viridis cytochrome c 2 (cycA) has been introduced on a broad host range vector into Paracoccus denitrificans , leading to high-level expression of the holo-cytochrome with the heme moiety covalently attached to the apoprotein. The cytochrome was demonstrated to reside in the periplasmic space of the host cell. In contrast to R. viridis , aerobic rather than anaerobic growth conditions led to higher production levels of the holo-cytochrome in P. denitrificans . This heterologous expression system provides a suitable genetic background for the functional expression and mutagenesis of polypeptides involved in bacterial photosynthesis, offering the possibility of detailed structural and functional investigation. |
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| Keywords: | Rhodopseudomonas viridis Paracoccus denitrificans Cytochrome c2 Cytochrome c synthesis Heme attachment Broad host range vector |
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