Abstract:A biosensor based on conductive poly(pyrrole‐co‐pyrrole‐2‐carboxylic acid) [Poly(Py‐co‐PyCOOH)] copolymer film coated gold electrode was developed for the quantitative phosphate determination. Enzyme pyruvate oxidase was immobilized chemically via the functional carboxylated groups of the copolymer. The potential to be applied which is deficiency of phosphate biosensor studies for precise phosphate detection was clarified by using differential pulse voltammetry technique. Performance of the sensing ability of … Show more
“…The carrier solution was composed of 20 mM pH 7.0 oxygenated citrate buffer containing 0.7 mg mL À 1 of LiClO 4 used as supporting electrolyte, 1 mM of magnesium (Mg + 2 ), 0.5 mM of pyruvate, 50 μM of TPP and 5 μM of FAD. Concentrations of Mg + 2 , pyruvate, TPP and FAD cofactors were optimized in our previous study [9]. After a stable background current was reached, analytes were injected into the flow-injection system using the injection valve and amperometric measurments were monitored online on the computer screen of the potentiostat.…”
Section: Apparatus and Detection Systemmentioning
confidence: 99%
“…Pyruvate oxidase, which belongs to the family of oxidoreductases, is an enzyme that catalyzes the oxidative decarboxylation of pyruvate, phosphate and oxygen to acetyl phosphate, carbon dioxide and hydrogen peroxide (equation 1) in the presence of cofactors such as FAD, TPP and Mg + 2 . TPP and FAD have an important place in the overall enzymatic reaction mechanism in presence of Mg + 2 [9]. The reaction mechanism proceeds as follows: After the ionization of the TPP C2-H, pyruvate reacts with the C2 carbanion in the active site, leading to the formation of 2-lactyl-TPP.…”
Section: Determination Of Working Potential For Phosphate Biosensormentioning
confidence: 99%
“…Meanwhile, the formed 2-hydroxyethyl-TPP is oxidized by FAD, leading to the formation of acetyl-TPP and two-electron reduced FADH 2 . The reduced form of FAD (FADH 2 ) is re-oxidized with molecular oxygen to form hydrogen peroxide and FAD, then acetyl-TPP is split apart phosphorolytically to acetyl phosphate and TPP C2 carbanion [9,32,33]. Rhodium nanoparticles acted as mediator for the electrocatalytic reduction of H 2 O 2 .…”
Section: Determination Of Working Potential For Phosphate Biosensormentioning
confidence: 99%
“…Even though the phosphate concentration of urine is measured by colorimetric analysis, the commonly used method is spectrophotometric at present [1,6]. These conventional methods have certain disadvantages such as tedious, time consuming, the use of health hazardous carcinogenic chemicals, requiring expertise and expensive instrumentation [4,9]. An alternative approach to the conventional method to overcome these shortcomings is biosensors with high selectivity and specificity to measure phosphate directly.…”
Section: Introductionmentioning
confidence: 99%
“…Interest in the development of phosphate biosensors for use in different related fields such as biological, environmental, food and agricultural has been continuing since the first report published by Guilbault and Nanjo in 1975 [7,10]. Although researchers have developed singleenzyme biosensors [9,[11][12][13], bi-enzyme biosensors [14][15][16][17], tri enzyme biosensors [18,19] for the over four decades, the simple, cost effective, more efficient and preferred one is pyruvate oxidase based single-enzyme phosphate biosensors [9]. Pyruvate oxidase catalyzes the oxidative decarboxylation of pyruvate, phosphate and oxygen to acetyl phosphate, carbon dioxide and hydrogen peroxide in the presence of cofactors such as flavin adenine dinucleotide (FAD), thiamine pyrophosphate chloride (TPP) and magnesium (Mg + 2 ).…”
A biosensor for phosphate determination with the flow‐injection system was developed using rhodium nanoparticles modified Poly(pyrrole‐co‐[1‐(2‐aminophenyl) pyrrole])/pyruvate oxidase. The biosensor showed a very wide linearity up to 70 mM phosphate concentration compared to previously reports, response time of 4 s., operational stability with a relative standard deviation of 0.009 % and accuracy of 99.4 %±0.949 at a flow rate of 2.0 Ml min.−1 at exactly −0.68 V. Detection limit were calculated to be 21±0.001 μM by preserving 81.1 % of its initial response at the end of 16th days. Artificial urine was analyzed without dilution to investigate biosensor performance.
“…The carrier solution was composed of 20 mM pH 7.0 oxygenated citrate buffer containing 0.7 mg mL À 1 of LiClO 4 used as supporting electrolyte, 1 mM of magnesium (Mg + 2 ), 0.5 mM of pyruvate, 50 μM of TPP and 5 μM of FAD. Concentrations of Mg + 2 , pyruvate, TPP and FAD cofactors were optimized in our previous study [9]. After a stable background current was reached, analytes were injected into the flow-injection system using the injection valve and amperometric measurments were monitored online on the computer screen of the potentiostat.…”
Section: Apparatus and Detection Systemmentioning
confidence: 99%
“…Pyruvate oxidase, which belongs to the family of oxidoreductases, is an enzyme that catalyzes the oxidative decarboxylation of pyruvate, phosphate and oxygen to acetyl phosphate, carbon dioxide and hydrogen peroxide (equation 1) in the presence of cofactors such as FAD, TPP and Mg + 2 . TPP and FAD have an important place in the overall enzymatic reaction mechanism in presence of Mg + 2 [9]. The reaction mechanism proceeds as follows: After the ionization of the TPP C2-H, pyruvate reacts with the C2 carbanion in the active site, leading to the formation of 2-lactyl-TPP.…”
Section: Determination Of Working Potential For Phosphate Biosensormentioning
confidence: 99%
“…Meanwhile, the formed 2-hydroxyethyl-TPP is oxidized by FAD, leading to the formation of acetyl-TPP and two-electron reduced FADH 2 . The reduced form of FAD (FADH 2 ) is re-oxidized with molecular oxygen to form hydrogen peroxide and FAD, then acetyl-TPP is split apart phosphorolytically to acetyl phosphate and TPP C2 carbanion [9,32,33]. Rhodium nanoparticles acted as mediator for the electrocatalytic reduction of H 2 O 2 .…”
Section: Determination Of Working Potential For Phosphate Biosensormentioning
confidence: 99%
“…Even though the phosphate concentration of urine is measured by colorimetric analysis, the commonly used method is spectrophotometric at present [1,6]. These conventional methods have certain disadvantages such as tedious, time consuming, the use of health hazardous carcinogenic chemicals, requiring expertise and expensive instrumentation [4,9]. An alternative approach to the conventional method to overcome these shortcomings is biosensors with high selectivity and specificity to measure phosphate directly.…”
Section: Introductionmentioning
confidence: 99%
“…Interest in the development of phosphate biosensors for use in different related fields such as biological, environmental, food and agricultural has been continuing since the first report published by Guilbault and Nanjo in 1975 [7,10]. Although researchers have developed singleenzyme biosensors [9,[11][12][13], bi-enzyme biosensors [14][15][16][17], tri enzyme biosensors [18,19] for the over four decades, the simple, cost effective, more efficient and preferred one is pyruvate oxidase based single-enzyme phosphate biosensors [9]. Pyruvate oxidase catalyzes the oxidative decarboxylation of pyruvate, phosphate and oxygen to acetyl phosphate, carbon dioxide and hydrogen peroxide in the presence of cofactors such as flavin adenine dinucleotide (FAD), thiamine pyrophosphate chloride (TPP) and magnesium (Mg + 2 ).…”
A biosensor for phosphate determination with the flow‐injection system was developed using rhodium nanoparticles modified Poly(pyrrole‐co‐[1‐(2‐aminophenyl) pyrrole])/pyruvate oxidase. The biosensor showed a very wide linearity up to 70 mM phosphate concentration compared to previously reports, response time of 4 s., operational stability with a relative standard deviation of 0.009 % and accuracy of 99.4 %±0.949 at a flow rate of 2.0 Ml min.−1 at exactly −0.68 V. Detection limit were calculated to be 21±0.001 μM by preserving 81.1 % of its initial response at the end of 16th days. Artificial urine was analyzed without dilution to investigate biosensor performance.
A novel nanostructured platform for pyruvate oxidase biosensors comprises poly(neutral red) (PNR) prepared by electropolymerization of NR in ethaline deep eutectic solvent (DES) with acid dopant, on a multiwalled carbon nanotubes (MWCNT) modified glassy carbon electrode (GCE). Characterization was by cyclic voltammetry and electrochemical impedance spectroscopy and morphology was examined by scanning electron micro-scopy. Ascorbate and H 2 O 2 gave a better response at PNR DES /GCE than at PNR aq /GCE. Biosensors for pyruvate and phosphate, immobilizing pyruvate oxidase onto PNR DES /MWCNT/GCE enabled selective determination of pyruvate and phosphate, with micromolar limits of detection. Pyruvate was determined in onion samples and phosphate in water samples. Keywords: Ethaline deep eutectic solvent • poly(neutral red) • MWCNT • pyruvate oxidase • phosphate [a] R.
A continuous flow enzymatic fuel cell (EFC) based on poly(pyrrole-co-pyrrole-2-carboxylic acid) film was used for sustainable energy generation. The system parameters were optimized to generate maximum power. Optimum cycle number for electropolymerization was found to be 12.5 for bioanode, and 20 for biocathode.The best performance was reached at pH 7, 1 ml min À1 flow rate and cell potential of 0.51 V. A power density of 0.36 μW cm À2 in 10 min was generated with 0.86 μg glucose dehydrogenase and 0.35 μg bilirubin oxidase enzyme quantity by using 100 mmol L À1 glucose. The EFC performance was investigated with municipal sewage, and the system generated a power density of 0.85 μW cm À2 . Results showed that the proposed EFC could be successfully used at waste utilization, especially for the wastewater with glycolytic content.
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