The implementation of the FluoRox principle into a working prototype FluoRox sensor requires four steps. The first step consists of the development of the element that actually performs the detection, i.e., the protein/enzyme. We have chosen three enzymes,
- nitrite reductase (NiR, from Alcaligenes faecalis),
- cytochrome P450 BM3 (P450, from Bacillus megatherium) and
- laccase (LAC, from Streptomyces coelicolor, Bacillus subtilis and Trametes versicolor)
and their reaction partners for incorporation into a FluoRox sensor. These will lead to prototypes of sensors that stand for three major types of application: environmental monitoring (NiR), medical and pharmaceutical diagnostics (P450, NiR), industrial bulk and fine-chemical processes (LAC). The second step involves electrode preparation and immobilization of the enzyme/protein onto it. The third step is electrochemical characterization of the construct (electrode with immobilized protein/enzyme). The fourth and last step comprises construction and testing of a prototype FluoRox sensor. Expertise, strains and equipment are available in the consortium to secure batchwise production of purified enzymes at the 10-100 mg (NiR, LAC) or the 1-10 mg scale (P450).