A new pyruvate oxidase biosensor based on 3-mercaptopropionic acid/6-aminocaproic acid modified gold electrode

Figures & data

Scheme 1. The reaction mechanism between the redox center of POX and the electrode surface.

Figure 1. Cyclic voltammograms of bare AuE (a), AuE/3-MPA/6-ACA (b), AuE/3-MPA/6-ACA/POX/GA in the absence of pyruvate (c) and AuE/ 3-MPA/6-ACA/POX/GA in the presence of pyruvate (d) in phosphate buffer (pH 6.0, 50 mM containing 0.1 M KCl and 5.0 mM K₃Fe(CN)₆) at a scan rate of 50 mVs^{− 1} vs. Ag/AgCl.

Figure 2. DP Voltammograms obtained for different concentration of pyruvate; a (0 μM), b (2.5 μM), c (10 μM), d (20 μM), e (30 μM), f (40 μM), g (50 μM). Inset shows the relationship between catalytic currents and pyruvate concentrations. Conditions: pH 6.0, 50 mM phosphate buffer (containing 0.1 M KCl and 5.0 mM K₃Fe(CN)₆) at 30°C. The percentage of glutaraldehyde and the activity of pyruvate oxidase were kept constant to be 2.5% and 0.356 UmL^{− 1}, respectively.

Figure 3. Current–time curves obtained with the biosensor upon successive addition of 1 mL 30 μM pyruvate. Inset shows the relationship between catalytic currents and pyruvate concentrations. Conditions: pH 6.0, 50 mM phosphate buffer (containing 0.1 M KCl and 5.0 mM K₃Fe(CN)₆) at the applied potential of 30 mV. The percentage of glutaraldehyde and the activity of pyruvate oxidase were kept constant to be 2.5% and 0.356 UmL^{− 1}, respectively.

Table I. Comparison of performances of some biosensors developed for determination of pyruvate.

Biosensor	Detection limit	Linear range	Response time	References
Chemiluminescent FIA/POX Biosensor	0.16 μM	0.32–32 μM	–	Nakamura et al. (1997)
Dissolved oxygen electrode (DOE) glutaraldehyde/POX	0.025 μM	0.025–0.5 μM	2 min	Akyilmaz and Yorganci (2008)
(GCE)/poly-TTCA (Nano-CP)/covalent immobilization of POX	0.3 μM	1.0–100 μM	6 min	Rahman et al. (2006)
POX/poly(mercapto-p-benzoquinone)	1.0 μM	1.0–2000 μM	–	Arai et al. (1999)
3-MPA/6-ACA/POX/GA modified AuE	1.87 μM (DP)	2.5–50 μM	6 s	Present work
0.56 μM (Amp)	1.0–10 μM	2 s		Present work

FIA, Flow injection analysis; POX, Pyruvate oxidase; GCE, Glassy carbon electrode; TTCA, 5,2:5,2-terthiophene-3-carboxylic acid; 3-MPA, 3-Mercapto propionic acid; 6-ACA, 6-Amino caproic acid; GA, Glutaraldehyde.

Nakamura H, Ikebukuro K, McNiven S, Karube I, Yamamoto H, Hayashi K, et al. 1997. A chemiluminescent FIA biosensor for phosphate ion monitoring using pyruvate ox dase. Biosens Bioelectron. 12:959.

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Akyilmaz E, Yorganci E. 2008. A novel biosensor based on activation effect of thiamine on the activity of pyruvate oxidase. Biosens Bioelectron. 23:1874.

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Rahman AM, Park DS, Chang SC, McNeil CJ, Shim YB. 2006. The biosensor based on the pyruvate oxidase modified conducting polymer for phosphate ions determinations. Biosens Bioelectron. 21:1116.

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Arai G, Noma T, Habu H, Yasumori I. 1999. Pyruvate sensor based on pyruvate oxidase immobilized in a poly(mercapto-p- benzoquinone) film. J Electroanal Chem. 464:143.

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