Conducting Polymer Based Enzyme Electrodes Fabricated by Invertase and Polyphenol Oxidase

Öz

Novel carbon fiber enzyme electrodes were constructed and evaluated. Fabrication of electrodes was performed via electrochemical deposition of a conducting matrice composed of polythiophene and polypyrrole (PTh–PPy) onto carbon fiber substrates. The enzyme was entrapped into the matrix during electropolymerization. Resultant biosensors represented higher kinetic parameters, Vmax and Km, in comparison to PPy matrice, which are 2.471 ± 0.150 mol min1- electrode1- and 30.60 ± 5.30 mM for invertase, 0.056 ± 0.012 mol min1- electrode1- and 842.00 ± 37.50 mM for polyphenol oxidase respectively. Optimum pH and temperature of the immobilized enzyme within PTh–PPy composite indicates that this matrice provides a more protective environment. The detection limit (LOD) of polyphenols was obtained as 0.037 mg mL1-. Polyphenol oxidase enzyme electrodes were proved to be used for the determination of polyphenolic substances in real samples and the results were confirmed by the Folin-Ciocalteau method

Anahtar Kelimeler

• Carbon Fiber Electrode
• Conducting Polymer
• Enzyme Immobilization
• Polyphenol Oxidase

Kaynakça

1. [1] Fei J., Wu K., Wang F., Hu S., 2005. Glucose nanosensors based on redox polymer/glucose oxidase modified carbon fiber nanoelectrodes. Talanta, 65, pp. 918-924.
2. [2] Taylor I.M., Robbins E.M., Catt K.A., Cody P.A., Happe C.L., Cui X.T., 2017. Enhanced dopamine detection sensitivity by PEDOT/graphene oxide coating on in vivo carbon fiber electrodes. Biosensors and Bioelectronics, 89, pp. 400-410.
3. [3] Salazar P., Martin M., O’Neill R.D., Gonzalez-Mora J.L., 2016. Glutamate microbiosensors based on Prussian Blue modified carbon fiber electrodes for neuroscience applications: In vitro characterization. Sensors and Actuators B: Chemical, 235, pp. 117-125.
4. [4] Ferreira N.R., Ledo A., Laranjinha J., Gerhardt G.A., Barbosa R.M., 2018. Simultaneous measurements of ascorbate and glutamate in vivo in the rat brain using carbon fiber nanocomposite sensors and microbiosensor arrays. Bioelectrochemistry, 121, pp. 142-150.
5. [5] Kim J.H., Cho S., Bae T.S., Lee Y.S., 2014. Enzyme biosensor based on an N-doped activated carbon fiber electrode prepared by a thermal solid-state reaction. Sensors and Actuators B: Chemical, 197, pp. 20-27.
6. [6] Yuan C.J., Wang C.L., Wu T.Y., Hwang K.C., Chao W.C., 2011. Fabrication of a carbon fiber paper as the electrode and its application toward developing a sensitive unmediated amperometric biosensor. Biosensors and Bioelectronics, 26, pp. 2858-2863.
7. [7] Nadaroglu H., Mosber G., Alayli Gungor A., Adıguzel G., 2019. Biodegradation of some azo dyes from wastewater with laccase from Weissella viridescens LB37 immobilized on magnetic chitosan nanoparticles. Journal of Water Process Engineering, 31, 100866.
8. [8] Nambiar S., Yeow J.T.W., 2011. Conductive polymer-based sensors for biomedical applications. Biosensors and Bioelectronics, 26, pp. 1825-1832.

9. [9] Park C.S., Lee C., Kwon O.S., 2016. Conducting polymer based nanobiosensors. Polymers, 8, 249, pp. 1-18.
10. [10] Sarma A.K., Vatsyayan P., Goswami P., Minteer S.D., 2009. Recent advances in material science for developing enzyme electrodes. Biosensors and Bioelectronics, 24, pp. 2313-2322.
11. [11] Adeloju S.B., Moline A.N., 2001. Fabrication of ultra-thin polypyrrole-glucose oxidase film from supporting electrolyte-free monomer solution for potentiometric biosensing of glucose. Biosensors and Bioelectronics, 16, pp. 133-139.
12. [12] Besombes J.L., Cosnier S., Labbe P., 1997. Improvement of poly(amphiphilic pyrrole) enzyme electrodes via the incorporation of synthetic laponite-clay-nanoparticles. Talanta, 44, pp. 2209-2215.
13. [13] Borochov-Neori H., Judeinstein S., Tripler F., Harari M., Greenberg A., Shomer I., Holland D., 2009. Seasonal and cultivar variations in antioxidant and sensory quality of pomegranate (Punica granatum L.) fruit. Journal of Food Composition and Analysis, 22, pp. 189-195.
14. [14] Tagliazucchi V., Verzelloni E., Bertolini D., Angela C., 2010. In vitro bio-accessibility and antioxidant activity of grape polyphenols. Food Chemistry, 120, pp. 599-606.
15. [15] Del Rio D., Costa L.G., Lean M.E.G., Crozier A., 2010. Polyphenols and health: What compounds are involved? Nutrition, Metabolism and Cardiovascular Diseases, 20, pp. 1-6.
16. [16] Apetrei C., Rodriguez-Mendez M.L., De Saja J.A., 2011. Amperometric tyrosinase based biosensor using an electropolymerized phosphate-doped polypyrrole film as an immobilization support. Application for detection of phenolic compounds. Electrochimica Acta, 56, pp. 8919- 8925.
17. [17] Nelson N., 1944. A photometric adaptation of Somogyi method for the determination of glucose. Journal of Biological Chemistry, 153, pp. 375-380.
18. [18] Pifferi P.G., Baldassari L., 1973. A spectrophotometric method for the determination of catecholase activity of tyrosinase by Besthorn’s hydrazone. Analytical Biochemistry, 52, pp. 325-335.
19. [19] Lineweaver H., Burk D., 1934. The determination of enzyme dissociation constants. Journal of the American Chemical Society, 56, pp. 658-666.
20. [20] Singleton V.L., Orthofer R., Lamuela-Raventos R., 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Oxidants and Antioxidants Part A Book Series: Methods in Enzymology, Volume 299, 152-178, 1st ed. Academic Press, United States.
21. [21] Karabacak G., 2019. İnvertaz ve polifenol oksidaz enzimleriyle oluşturulan karbon fiber enzim elektrotları. MSc Thesis, Graduate Enstitute, Karabük University, Karabük.
22. [22] Böyükbayram A.E., Kıralp S., Toppare L., Yağcı Y., 2006. Preparation of biosensors by immobilization of polyphenol oxidase in conducting copolymers and their use in determination of phenolic compounds in red wine. Bioelectrochemistry, 69, pp. 164-171.
23. [23] Alkan S., Toppare L., Yağci Y., Hepuzer Y., 1999. Immobilization of invertase in conducting thiophene-capped poly(methyl methacrylate)/polypyrrole matrices. Journal of Biomaterials Science, Polymer Edition, 10, pp. 1223-1235.
24. [24] Kiralp S., Toppare L., Yagci Y., 2003. Immobilization of polyphenol oxidase in conducting copolymers and determination of phenolic compounds in wines with enzyme electrodes. International Journal of Biological Macromolecules, 33, pp. 37-41.
25. [25] Erginer R., Toppare L., Alkan S., Bakır U., 2000. Immobilization of invertase in functionalized copolymer matrices. Reactive and Functional Polymers, 45, pp. 227-233.