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

An enzymatic biosensor based on nitric oxide reductase (NOR; purified from Marinobacter hydrocarbonoclasticus) was developed for nitric oxide (NO) detection. The biosensor was prepared by deposition onto a pyrolytic graphite electrode (PGE) of a nanocomposite constituted by carboxylated single-walle...

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Published in:Bioelectrochemistry (Amsterdam, Netherlands) Netherlands), 2019-06, Vol.127, p.76-86
Main Authors: Gomes, Filipa O., Maia, Luísa B., Loureiro, Joana A., Pereira, Maria Carmo, Delerue-Matos, Cristina, Moura, Isabel, Moura, José J.G., Morais, Simone
Format: Article
Language:English
Subjects:
Arginine
Ascorbic acid
Bilayers
Biomimetics
Biosensors
Direct electron transfer
Electrochemistry
Electron transfer
Enzymatic biosensor
Fourier transforms
Infrared spectroscopy
Nanocomposites
Nanotechnology
Nanotubes
Nitric oxide
Nitric oxide reductase
Phospholipids
Polyethylene glycol
Pyrolytic graphite
Reproducibility
Scanning electron microscopy
Selectivity
Single wall carbon nanotubes
Sodium
Sodium nitrate
Sodium nitrite
Stability
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Summary:An enzymatic biosensor based on nitric oxide reductase (NOR; purified from Marinobacter hydrocarbonoclasticus) was developed for nitric oxide (NO) detection. The biosensor was prepared by deposition onto a pyrolytic graphite electrode (PGE) of a nanocomposite constituted by carboxylated single-walled carbon nanotubes (SWCNTs), a lipidic bilayer [1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-di-(9Z-octadecenoyl)-3-trimethylammonium-propane (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol (DSPE-PEG)] and NOR. NOR direct electron transfer and NO bioelectrocatalysis were characterized by several electrochemical techniques. The biosensor development was also followed by scanning electron microscopy and Fourier transform infrared spectroscopy. Improved enzyme stability and electron transfer (1.96 × 10−4 cm.s−1 apparent rate constant) was obtained with the optimum SWCNTs/(DOPE:DOTAP:DSPE-PEG)/NOR) ratio of 4/2.5/4 (v/v/v), which biomimicked the NOR environment. The PGE/[SWCNTs/(DOPE:DOTAP:DSPE-PEG)/NOR] biosensor exhibited a low Michaelis-Menten constant (4.3 μM), wide linear range (0.44–9.09 μM), low detection limit (0.13 μM), high repeatability (4.1% RSD), reproducibility (7.0% RSD), and stability (ca. 5 weeks). Selectivity tests towards L-arginine, ascorbic acid, sodium nitrate, sodium nitrite and glucose showed that these compounds did not significantly interfere in NO biosensing (91.0 ± 9.3%–98.4 ± 5.3% recoveries). The proposed biosensor, by incorporating the benefits of biomimetic features of the phospholipid bilayer with SWCNT's inherent properties and NOR bioelectrocatalytic activity and selectivity, is a promising tool for NO. •A biosensor based on M. hydrocarbonoclasticus nitric oxide reductase was developed.•SWCNTs/lipidic bilayer nanocomposite improved enzyme catalytic activity and stability.•The biosensor exhibited an apparent rate constant of 1.96 × 10−4 cm.s−1.•The biosensor showed a NO detection limit of 0.13 μM and wide linear range.•The biosensor retained 83.5% of its initial response after five weeks.
ISSN:1567-5394
1878-562X
DOI:10.1016/j.bioelechem.2019.01.010