The cytochrome complex SoxXA of Paracoccus pantotrophus is produced in Escherichia coli and functional in the reconstituted sulfur-oxidizing enzyme system.
The heterodimeric c-type cytochrome complex SoxXA of Paracoccus pantotrophus was produced in Escherichia coli. The soxX and soxA genes, separated by two genes in the sox gene cluster of P. pantotrophus, were fused with ribosome binding sites optimal for E. coli and combined to give soxXA in pRD133.27. The cytochrome complex SoxXA was produced in E. coli M15 containing pRD133.27, pREP4 encoding the Lac repressor and plasmid pEC86, carrying essential cytochrome c maturation genes. SoxX and SoxA were formed in a ratio of about 2.5:1. SoxA appeared to be unstable when not complexed with SoxX. The cytochrome complex SoxXA, purified to homogeneity from periplasmic extracts of E. coli M15 (pRD133.27, pREP4, pEC86), exhibited identical biochemical and biophysical properties as compared to SoxXA of P. pantotrophus. Moreover, this cytochrome complex was shown to be equally catalytically active with respect to rates and reactivity with different sulfur substrates in the reconstituted sulfur-oxidizing enzyme system using homogeneous Sox-proteins of P. pantotrophus. Homogeneous SoxX was catalytically inactive.
Rother
Dagmar
D
Lehrstuhl für Technische Mikrobiologie, Fachbereich Chemietechnik, Universität Dortmund, Dortmund, Germany.
Friedrich
Cornelius G
CG
D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005838:Genotype; D005810:Multigene Family; D010230:Paracoccus / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D021122:Protein Subunits; D011994:Recombinant Proteins