Browsing by Author "Cordas, Cristina"
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- Electroanalytical characterization of the direct Marinobacter hydrocarbonoclasticus nitric oxide reductase-catalysed nitric oxide and dioxygen reductionPublication . Gomes, Filipa O.; Maia, Luísa B.; Cordas, Cristina; Moura, Isabel; Delerue-Matos, Cristina; Moura, José J.G.; Morais, SimoneUnderstanding the direct electron transfer processes between redox proteins and electrode surface is fundamental to understand the proteins mechanistic properties and for development of novel biosensors. In this study, nitric oxide reductase (NOR) extracted from Marinobacter hydrocarbonoclasticus bacteria was adsorbed onto a pyrolytic graphite electrode (PGE) to develop an unmediated enzymatic biosensor (PGE/NOR)) for characterization of NOR direct electrochemical behaviour and NOR electroanalytical features towards NO and O2. Square-wave voltammetry showed the reduction potential of all the four NOR redox centers: 0.095 ± 0.002, -0.108 ± 0.008, -0.328 ± 0.001 and -0.635 ± 0.004 V vs. SCE for heme c, heme b, heme b3 and non-heme FeB, respectively. The determined sensitivity (-4.00 × 10-8 ± 1.84 × 10-9 A/μM and - 2.71 × 10-8 ± 1.44 × 10-9 A/μM for NO and O2, respectively), limit of detection (0.5 μM for NO and 1.0 μM for O2) and the Michaelis Menten constant (2.1 and 7.0 μM for NO and O2, respectively) corroborated the higher affinity of NOR for its natural substrate (NO). No significant interference on sensitivity towards NO was perceived in the presence of O2, while the O2 reduction was markedly and negatively impacted (3.6 times lower sensitivity) by the presence of NO. These results clearly demonstrate the high potential of NOR for the design of innovative NO biosensors.
- Nitric Oxide Detection Using Electrochemical Third-generation Biosensors - Based on Heme Proteins and PorphyrinsPublication . Gomes, Filipa O.; Maia, Luísa B.; Cordas, Cristina; Delerue-Matos, Cristina; Moura, Isabel; Moura, José J. G.; Morais, SimoneNitric oxide radical (NO) is a signalling molecule involved in virtually all forms of life. Its relevance has been leading to the development of different analytical methodologies to assess the temporal and spatial fluxes of NO under the complex biological milieu. Third‐generation electrochemical biosensors are promising tools for in loco and in vivo NO quantification and, over the past years, heme proteins and porphyrins have been used in their design. Since there are some limitations with the biorecognition element directly adsorbed onto the electrode surface, nanomaterials (carbon nanotubes, gold nanoparticles, etc.) and polymers (cellulose, chitosan, nafion®, polyacrylamide, among others) have been explored to achieve high kinetics and better biosensor performance. In this review, a broad overview of the field of electrochemical third‐generation biosensors for NO electroanalysis is presented, discussing their main characteristics and aiming new outlooks and advances in this field.
