Exploiting immobilized engineered cytochrome c in bioelectronic sensing devices

In recent years, redox metalloproteins have increasingly beenperceived as good candidates to serve as basic functional units ofnano-structured biomolecular surfaces at the heart ofinorganic/biological electronic devices such as biosensors andbiotransistors. Our approach was to engineer theheme-containing cytochrome c, which withstands extremeconditions in terms of pH, temperature and the presence ofnonaqueous solvents, turning it into a chimeric peroxidase. Wefound that the Met80Ala cytc variant immobilized on a variety of self-assembled monolayers on a gold electrode shows a remarkable ability to catalytically reduce O2 in alarge pH range (from 5 to 11.5). This behavior indicates thatengineered five-coordinate heme-containing cytcs are promisingcandidates for the amperometric reductive biosensing of molecular oxygen, also opening the way to the biosensing of hydrogen peroxide and organic hydroperoxides, of potential use to monitor the cellular oxidative stress.