Artículos
Detección electroquímica de peróxido de hidrógeno usando peroxidasa de pasto Guinea (Panicum maximum) inmovilizada sobre electrodos serigrafiados de puntos cuánticos
Publicado 2020-03-11
Palabras clave
- Biosensor,
- Peroxidasa,
- Pasto Guinea,
- Peróxido de Hidrógeno,
- Puntos Cuánticos.
Cómo citar
Guarín, P., Cano, H. J., & Castillo, J. J. (2020). Detección electroquímica de peróxido de hidrógeno usando peroxidasa de pasto Guinea (Panicum maximum) inmovilizada sobre electrodos serigrafiados de puntos cuánticos. Revista ION, 32(2), 67–76. https://doi.org/10.18273/revion.v32n2-2019007
Resumen
Los biosensores electroquímicos son herramientas analíticas de rápida y confiable respuesta que han adquirido especial interés en los últimos años gracias a la posibilidad de integrar biomoléculas con electrodos hechos a base de materiales nanométricos. En este trabajo se desarrolló un biosensor electroquímico para detección de peróxido de hidrógeno (H2O2) usando peroxidasa de pasto Guinea (PPG) inmovilizada sobre electrodos serigrafiados de puntos cuánticos (ESPC). La PPG fue aislada y parcialmente purificada a partir de hojas de pasto Guinea con una actividad específica de 602 Umg-1. Posteriormente, la PPG fue inmovilizada sobre la superficie del ESPC mediante adsorción física y el estudio del comportamiento electroquímico fue llevado a cabo mediante voltamperometría cíclica y cronoamperometría. La PPG reveló una pareja bien definida de señales redox a 17mV/-141mV correspondientes al proceso redox del grupo hemo (Fe2+/Fe3+) de las peroxidasas. La reducción bioelectrocatalítica del peróxido de hidrógeno se observó a un potencial redox de -645 mV vs Ag. Este proceso fue controlado por difusión de las especies en la superficie del electrodo en un rango de velocidad de barrido lineal de 50-500 mV/s. La cronoamperometría permitió la construcción de curvas de calibración entre la corriente de reducción y la concentración del H2O2 para la determinación de parámetros analíticos como sensibilidad, rango lineal y nivel mínimo de detección. El desarrollo de este biosensor amperométrico se convierte en un paso preliminar para la construcción de un dispositivo portátil y de respuesta rápida para el análisis de H2O2 en muestras de interés ambiental y biomédico.
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Referencias
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[23] Ramírez EA, Granero AM, Zon MA, Fernández H. Development of an Amperometric Biosensor Based on Peroxidases from Brassica napus for the Determination of Ochratoxin a Content in Peanut Samples. J Biosens Bioelectron S. 2011;3:1–6.
[24] Castillo J, Ferapontova E, Hushpulian D, Tasca F, Tishkov V, Chubar T, et al. Direct electrochemistry and bioelectrocatalysis of H2O2 reduction of recombinant tobacco peroxidase on graphite. Effect of peroxidase single-point mutation on Ca2+ modulated catalytic activity. J Electroanal Chem. 2006;588(1):112–21.
[25] Ruzgas T, Csöregi E, Emnéus J, Gorton L, Marko-Varga G. Peroxidase-modified electrodes: Fundamentals and application. Anal Chim Acta. 1996;330(2–3):123–38.
[26] Xu S, Zhang X, Wan T, Zhang C. A third-generation hydrogen peroxide biosensor based on horseradish peroxidase cross-linked to multi-wall carbon nanotubes. Microchim Acta. 2010;172(1–2):199–205.
[27] Zhang H-L, Zou X-Z, Lai G-S, Han D-Y, Wang F. Direct electrochemistry of hemoglobin immobilized on carhon-coated iron nanoparticles for amperometric detection of hydrogen peroxide. Electroanalysis. 2007;19(18):1869–74.
[28] Moyo M, Okonkwo JO, Agyei NM. A novel hydrogen peroxide biosensor based on adsorption of horseradish peroxidase onto a nanobiomaterial composite modified glassy carbon electrode. Electroanalysis. 2013;25(8):1946–54.
[29] Wang Z, Xu Q, Wang H-Q, Yang Q, Yu J-H, Zhao Y-D. Hydrogen peroxide biosensor based on direct electron transfer of horseradish peroxidase with vapor deposited quantum dots. Sensors Actuators B Chem. 2009;138(1):278–82.
[30] Yin H, Ai S, Shi W, Zhu L. A novel hydrogen peroxide biosensor based on horseradish peroxidase immobilized on gold nanoparticles–silk fibroin modified glassy carbon electrode and direct electrochemistry of horseradish peroxidase. Sensors Actuators B Chem. 2009;137(2):747–53.
[31] Kang XB, Pang GC, Liang XY, Wang M, Liu J, Zhu WM. Study on a hydrogen peroxide biosensor based on horseradish peroxidase/GNPs-thionine/chitosan. Electrochim Acta. 2012;62:327–34.
[32] Ma L, Yuan R, Chai Y, Chen S. Amperometric hydrogen peroxide biosensor based on the immobilization of HRP on DNA-silver nanohybrids and PDDA-protected gold nanoparticles. J Mol Catal B Enzym. 2009;56(4):215–20.
[33] Koposova E, Shumilova G, Ermolenko Y, Kisner A, Offenhäusser A, Mourzina Y. Direct electrochemistry of cyt c and hydrogen peroxide biosensing on oleylamine- and citrate-stabilized gold nanostructures. Sensors Actuators B Chem. 2015;207:1045–52.
[34] Ahirwal GK, Mitra CK. Direct electrochemistry of horseradish peroxidase-gold nanoparticles conjugate. Sensors. 2009;9(2):881–94.
[35] Yagati A, Min J, Cho S. Electrosynthesis of ERGO-NP nanocomposite films for bioelectrocatalysis of horseradish peroxidase towards H2O2. J. Electrochem. Soc. 2014;166:133-40.
[36] Zhang Q, Qiao Y, Zhang L, Wu S, Zhou H, Song X. Direct electrochemistry and electrocatalysis of horseradish peroxidase immobilized on water soluble sulfonated graphene film via self‐assembly. Electroanalysis. 2011;23:900-6.
[37] Farzana S, Ganesh V, Berchmans S. A sensing platform for direct electron transfer study of horseradish peroxidase. J. Electrochem. Soc. 2013;160:573-80.
[2] Shi L, Liu X, Niu W, Li H, Han S, Chen J, et al. Hydrogen peroxide biosensor based on direct electrochemistry of soybean peroxidase immobilized on single-walled carbon nanohorn modified electrode. Biosens Bioelectron. 2009;24(5):1159–63.
[3] Wang Y, Du J, Li Y, Shan D, Zhou X, Xue Z, et al. A amperometric biosensor for hydrogen peroxide by adsorption of horseradish peroxidase onto single-walled carbon nanotubes. Colloids Surf B Biointerfaces. 2012;90:62–7.
[4] Sakharov IY, Vesgac B MK, Galaev IY, Sakharova IV, Pletjushkina OY. Peroxidase from leaves of royal palm tree Roystonea regia: Purification and some properties. Plant Sci. 2001;161(5):853–60.
[5] Centeno DA, Solano XH, Castillo JJ. A new peroxidase from leaves of guinea grass (Panicum maximum): A potential biocatalyst to build amperometric biosensors. Bioelectrochemistry. 2017;116:33-8.
[6] Orduz AE, Gutiérrez JA, Blanco SI, Castillo JJ, Salcedo-reyes JC. Amperometric detection of triclosan with screen-printed carbon nanotube electrodes modified with Guinea Grass (Panicum maximum) peroxidase. 2019;24(2):363–79.
[7] Vargas JA, Castillo JJ. Desarrollo de un prototipo de biosensor basado en peroxidasa de palma real (Roystonea regia) y nanotubos de péptidos inmovilizados sobre electrodos de oro para detección de peróxido de hidrógeno. Av en Quim. 2016;11(3):105–11.
[8] Villamizar EN, Ríos CA, Castillo JJ. A hydrogen peroxide biosensor based on the immobilization of the highly stable royal palm tree peroxidase (Roystonea regia) with chitosan and glutaraldehyde on screen-printed graphene electrodes. J Mex Chem Soc. 2016;60(3):135-40.
[9] Tian K, Nie F, Luo K, Zheng X, Zheng J. A sensitive electrochemiluminescence glucose biosensor based on graphene quantum dot prepared from graphene oxide sheets and hydrogen peroxide. J Electroanal Chem. 2017;801:162–70.
[10] Muthurasu A, Ganesh V. Horseradish Peroxidase Enzyme Immobilized Graphene Quantum Dots as Electrochemical Biosensors. Appl Biochem Biotechnol. 2014;174(3):945–59.
[11] Manan FAA, Hong WW, Abdullah J, Yusof NA, Ahmad I. Nanocrystalline cellulose decorated quantum dots based tyrosinase biosensor for phenol determination. Mater Sci Eng C. 2019;99:37–46.
[12] Tang J, Wang B, Wu Z, Han X, Dong S, Wang E. Lipid membrane immobilized horseradish peroxidase biosensor for amperometric determination of hydrogen peroxide. Biosens Bioelectron. 2003;18(7):867–72.
[13] Cinti S, Arduini F, Vellucci G, Cacciotti I, Nanni F, Moscone D. Carbon black assisted tailoring of Prussian Blue nanoparticles to tune sensitivity and detection limit towards H2O2 by using screen-printed electrode. Electrochem commun. 2014;47:63–6.
[14] Gatselou VA, Giokas DL, Vlessidis AG, Prodromidis MI. Rhodium nanoparticle-modified screen-printed graphite electrodes for the determination of hydrogen peroxide in tea extracts in the presence of oxygen. Talanta. 2015;134:482–7.
[15] Sun J, Fang H, Chen H. Immobilization of horseradish peroxidase on a self-assembled monolayer modified gold electrode for the detection of hydrogen peroxide. Analyst. 1998;123(6):1365–8.
[16] Chen W, Cai S, Ren QQ, Wen W, Zhao YDi. Recent advances in electrochemical sensing for hydrogen peroxide: A review. Analyst. 2012;137(1):49–58.
[17] Xu X, Liu S, Ju H. A Novel Hydrogen Peroxide Sensor via the Direct Electrochemistry of Horseradish Peroxidase Immobilized on Colloidal Gold Modified Screen-printed Electrode. Sensors. 2003;3(9):350–60.
[18] Chen S, Yuan R, Chai Y, Xu L, Wang N, Li X, et al. Amperometric hydrogen peroxide biosensor based on the immobilization of horseradish peroxidase (HRP) on the layer-by-layer assembly films of gold colloidal nanoparticles and toluidine blue. Electroanalysis. 2006;18:471–7.
[19] Uribe PA, Ortiz CC, Centeno DA, Castillo JJ, Blanco SI, Gutierrez JA. Self-assembled Pt screen printed electrodes with a novel peroxidase Panicum maximum and zinc oxide nanoparticles for H2O2 detection. Colloids Surfaces A Physicochem Eng Asp. 2019;561:18–24.
[20] Battistuzzi G, Bellei M, Bortolotti CA, Sola M. Redox properties of heme peroxidases. Arch Biochem Biophys. 2010;500(1):21–36.
[21] Belcarz A, Ginalska G, Kowalewska B, Kulesza P. Spring cabbage peroxidases - Potential tool in biocatalysis and bioelectrocatalysis. Phytochemistry. 2008;69(3):627–36.
[22] Brusova Z, Ferapontova E, Sakharov I, Magner E, Gorton L. Bioelectrocatalysis of Plant Peroxidases Immobilized on Graphite in Aqueous and Mixed Solvent Media. Electroanalysis. 2005;17(5–6):460–8.
[23] Ramírez EA, Granero AM, Zon MA, Fernández H. Development of an Amperometric Biosensor Based on Peroxidases from Brassica napus for the Determination of Ochratoxin a Content in Peanut Samples. J Biosens Bioelectron S. 2011;3:1–6.
[24] Castillo J, Ferapontova E, Hushpulian D, Tasca F, Tishkov V, Chubar T, et al. Direct electrochemistry and bioelectrocatalysis of H2O2 reduction of recombinant tobacco peroxidase on graphite. Effect of peroxidase single-point mutation on Ca2+ modulated catalytic activity. J Electroanal Chem. 2006;588(1):112–21.
[25] Ruzgas T, Csöregi E, Emnéus J, Gorton L, Marko-Varga G. Peroxidase-modified electrodes: Fundamentals and application. Anal Chim Acta. 1996;330(2–3):123–38.
[26] Xu S, Zhang X, Wan T, Zhang C. A third-generation hydrogen peroxide biosensor based on horseradish peroxidase cross-linked to multi-wall carbon nanotubes. Microchim Acta. 2010;172(1–2):199–205.
[27] Zhang H-L, Zou X-Z, Lai G-S, Han D-Y, Wang F. Direct electrochemistry of hemoglobin immobilized on carhon-coated iron nanoparticles for amperometric detection of hydrogen peroxide. Electroanalysis. 2007;19(18):1869–74.
[28] Moyo M, Okonkwo JO, Agyei NM. A novel hydrogen peroxide biosensor based on adsorption of horseradish peroxidase onto a nanobiomaterial composite modified glassy carbon electrode. Electroanalysis. 2013;25(8):1946–54.
[29] Wang Z, Xu Q, Wang H-Q, Yang Q, Yu J-H, Zhao Y-D. Hydrogen peroxide biosensor based on direct electron transfer of horseradish peroxidase with vapor deposited quantum dots. Sensors Actuators B Chem. 2009;138(1):278–82.
[30] Yin H, Ai S, Shi W, Zhu L. A novel hydrogen peroxide biosensor based on horseradish peroxidase immobilized on gold nanoparticles–silk fibroin modified glassy carbon electrode and direct electrochemistry of horseradish peroxidase. Sensors Actuators B Chem. 2009;137(2):747–53.
[31] Kang XB, Pang GC, Liang XY, Wang M, Liu J, Zhu WM. Study on a hydrogen peroxide biosensor based on horseradish peroxidase/GNPs-thionine/chitosan. Electrochim Acta. 2012;62:327–34.
[32] Ma L, Yuan R, Chai Y, Chen S. Amperometric hydrogen peroxide biosensor based on the immobilization of HRP on DNA-silver nanohybrids and PDDA-protected gold nanoparticles. J Mol Catal B Enzym. 2009;56(4):215–20.
[33] Koposova E, Shumilova G, Ermolenko Y, Kisner A, Offenhäusser A, Mourzina Y. Direct electrochemistry of cyt c and hydrogen peroxide biosensing on oleylamine- and citrate-stabilized gold nanostructures. Sensors Actuators B Chem. 2015;207:1045–52.
[34] Ahirwal GK, Mitra CK. Direct electrochemistry of horseradish peroxidase-gold nanoparticles conjugate. Sensors. 2009;9(2):881–94.
[35] Yagati A, Min J, Cho S. Electrosynthesis of ERGO-NP nanocomposite films for bioelectrocatalysis of horseradish peroxidase towards H2O2. J. Electrochem. Soc. 2014;166:133-40.
[36] Zhang Q, Qiao Y, Zhang L, Wu S, Zhou H, Song X. Direct electrochemistry and electrocatalysis of horseradish peroxidase immobilized on water soluble sulfonated graphene film via self‐assembly. Electroanalysis. 2011;23:900-6.
[37] Farzana S, Ganesh V, Berchmans S. A sensing platform for direct electron transfer study of horseradish peroxidase. J. Electrochem. Soc. 2013;160:573-80.