Nitric Oxide Detection Using Electrochemical Third‐generation Biosensors – Based on Heme Proteins and Porphyrins

Abstract

Nitric 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.

Biosensor for direct bioelectrocatalysis detection of nitric oxide using nitric oxide reductase incorporated in carboxylated single-walled carbon nanotubes/lipidic 3 bilayer nanocomposite

“…The NO reduction catalytic potential (defined as the midpoint potential of the observed electrocatalytic wave [58]; Fig. SM4, Supplementary Material) was evidenced at −0.69 ± 0.02 V, a value similar to the one reported for other heme-protein (hemoglobin and myoglobin) sensors [ [59], [60], [61]]. NOR can also catalyze the reduction of dioxygen, because it has similarities with the heme copper oxidases family, namely a similar catalytic center [62].…”

“…Values of 0.13 μM for limit of detection (LOD) and 0.44 μM for limit of quantification (LOQ) were estimated based on 3*Sy-intercept/slope and 10*Sy-intercept/slope, respectively, where Sy-intercept is the standard deviation of the y-intercept [73]. The main figures of merit compare favorably with those described for hemoglobin- [70,74] and cyt c-based NO sensors [61,69,75], as well as, for enzymatic biosensors based on lipidic bilayer (Table 1); the only two exceptions were one Tyr-and one acetylcholinesterase-based biosensor for phenolic compounds [25] and organophosphate pesticides (dichlorvos used as a model compound), respectively [29]. The proposed PGE/[SWCNTs/(DOPE:DOTAP:DSPE-PEG)/NOR] biosensor may be applicable to screen NO levels released from biological systems.…”

“…This low micromolar Km value also highlights the clear benefits of using an enzyme specific for NO (NOR) on the biosensor. With a couple of exceptions [71,72], most of the NO thirdgeneration biosensors (based on heme proteins and porphyrins [61,69,70]) display considerably higher Km values. Fig.…”

Third-generation electrochemical biosensor based on nitric oxide reductase immobilized in a multiwalled carbon nanotubes/1-n-butyl-3-methylimidazolium tetrafluoroborate nanocomposite for nitric oxide detection

“…Only these two studies were found concerning NO biosensors that include an IL and heme-proteins in their construction. The immunological function and the pathological effects of NO are associated with high nanomolar to low micromolar concentrations [67,68]. Thus, the attained figures of merit are appropriate to allow application of the developed biosensor to detect NO levels in real biological samples, namely those related with modulation of blood flow, cardiovascular pathologies, neurodegenerative diseases, among others [6,7,68].…”

Electrochemical Characterization and Bioelectrocatalytic H2O2 Sensing of Non‐Symbiotic Hexa‐Coordinated Sugar Beet Hemoglobins

“…In this context, of particular interest is adaptation of Hbs for bioelectrocatalytic sensing of ROS and reactive nitrogen species (RNS) and signal transduction. To now, numerous examples of Hb‐based electroanalysis of O 2 , H 2 O 2 , nitric oxide and nitrite have been reported …”