Functional expression and utilization of bacterial cytochrome C__-

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Cytochrome c__-_<552> gene was cloned from the total DNA of the cells of Hydrogen obacter thermophilus. The gene was shown to contain a signal sequence. As a next step, the gene without the signal sequence was prepared. When Escherichia coli was transformed with the plasmid containing this gene and was cultivated under nitrate respiration, it produced cytochrome c__-_<552> within cytoplasm fraction.Cytochrome c__-_<551> gene was also cloned from the total DNA of the cells of Pseudomonas aeruginosa. When Pseudomonas aeruginosa was transformed with the plasmid containig this gene, it produced cytochrome c__-_<551> within periplasm fraction. The production rate was as high as 0.7 % of the total protein of the cell-free extract. An attempt was made to produce cytochrome c__-_<551> by using the other microorganisms (Escherichia coli or Pseudomonas putida), however, successful production could not be obtained.Reactivity of the two proteins (cytochrome c__-_<552> and cytochrome c__-_<551>) toward electrode was also investigated. Both cytochromes were shown to react with Pt-electrode in a solution state. Cytochrome c__-_<552> protein could react with Au-electrode in the presence of promotor, however, in its absence reactivity was not observed.