A system for the heterologous expression of complex redox proteins in Rhodobacter capsulatus: characterisation of recombinant sulphite:cytochrome c oxidoreductase from Starkeya novella.
The phototrophic purple non-sulfur bacterium Rhodobacter capsulatus expresses a wide variety of complex redox proteins in response to changing environmental conditions. Here we report the construction and evaluation of an expression system for recombinant proteins in that organism which makes use of the dor promoter from the same organism. A generic expression vector, pDorEX, was constructed and used to express sulphite:cytochrome c oxidoreductase from Starkeya novella, a heterodimeric protein containing both molybdenum and haem c. The recombinant protein was secreted to the periplasm and its biochemical properties were very similar to those of the native enzyme. The pDorEX system therefore seems to be potentially useful for heterologous expression of multi-subunit proteins containing complex redox cofactors.
Kappler
Ulrike
U
Department of Microbiology and Parasitology, Centre for Metals in Biology, School of Molecular and Microbial Sciences, The University of Queensland, St. Lucia, Qld. 4072, Australia. u.kappler1@mailbox.uq.edu.au
McEwan
Alastair G
AG
D001483:Base Sequence; D002942:Circular Dichroism; D003579:Cytochrome Reductases / Q000302:isolation & purification / Q000378:metabolism*; D017931:DNA Primers; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D016354:Rhodobacter capsulatus / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D050902:Sulfite Dehydrogenase; D013855:Thiobacillus / Q000378:metabolism*
D017931:DNA Primers; D011994:Recombinant Proteins; D003579:Cytochrome Reductases; D050902:Sulfite Dehydrogenase