Corresponding Author

Song-qin LIU(liusq@seu.edu.cn)


The process of drug metabolism plays an important role in drug efficacy and toxicity in vivo. Thus, the development of cheap, continent, rapid and high-throughput method for drug metabolism studies has great guiding significance for the design of new drugs, the determination of drug dosage and the detection of clinical drugs. Since the key role played by Cytochrome P450 (CYP450) in phase-I drug reaction, the constructed CYP450 enzyme biosensor can be used for the initial screening of drugs. It is found that the replacement of coenzyme NADPH with an electrode is to provide two electrons demanded in the catalytic reaction. Furthermore, the assembly methods and electrode materials have a close relationship with the detection performance of the constructed CYP450 biosensors. In this review, the construction methods of CYP450 electrochemical biosensor and their applications in drug metabolism are summarized. Particularly, future study and prospect of development are envisioned.

Graphical Abstract


cytochrome p450, drug metabolism, electrochemistry, biosensors, assembly methods

Publication Date


Online Available Date


Revised Date


Received Date



[1] Guengerich F P. Cytochrome P450 enzymes in the generation of commercial products[J]. Nature reviews-Drug discovery, 2002, 1(5): 359-366.
[2] Schneider E, Clark D S. Cytochrome P450 (CYP) enzymes and the development of CYP biosensors[J]. Biosensors & Bioelectronics, 2013, 39(1): 1-13.
[3] Bistolas N, Wollenberger U, Jung C, et al. Cytochrome P450 biosensors—a review[J]. Biosensors & Bioelectronics, 2005, 20(12): 2408-2423.
[4] Krishnan S, Schenkman J B, Rusling J F. Bioelectronic delivery of electrons to cytochrome P450 enzymes[J]. Journal of Physical Chemistry B, 2011, 115(26): 8371-8380.
[5] Estabrook R W, Faulkner K M, Seth M S, et al. Application of electrochemistry for P450-catalyzed reactions[J]. Method in Enzymology, 1996, 272: 44-51.
[6] Reipa V, Mayhew M P, Vilker V L. A direct electrode-driven P450 cycle for biocatalysis[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(25): 13554-13558.
[7] Wirtz M, Klucik J, Rivera M. Ferredoxin-mediated electrocatalytic dehalogenation of haloalkanes by Cytochrome P450cam[J]. Journal of the American Chemical Society, 2000, 122(6): 1047-1056.
[8] Kazlauskaite J, Westlake A C G, Wong L, et al. Direct electrochemistry of cytochrome P450cam[J]. Chemical Communications, 1996, 18: 2189-2190.
[9] Lo K K, Wong L, Hill H A O. Surface-modified mutants of cytochrome CYP101 enzymatic properties and electrochemistry[J]. FEBS Letters, 1999, 451: 342-346.
[10] Fantuzzi A, Fairhead M, Gilardi G. Direct electrochemistry of immobilized human cytochrome P450 2E1[J]. Journal of the American Chemical Society, 2004, 126(16): 5040-5041.
[11] Decher G, Hong J D, Schmitt J. Buildup of ultrathin multilayer films by a self-assembly process: III. Consecutively alternating adsorption of anionic and cationic polyelectrolytes on charged surfaces[J]. Thin Solid Films, 1992, 210(2): 831-835.
[12] Lvov Y, Decher G, Moehwald H. Assembly, structural characterization, and thermal behavior of layer-by-layer deposited ultrathin films of poly(vinyl sulfate) and poly(allylamine)[J]. Langmuir, 1993, 9(2): 481-486.
[13] Ariga K, Kunitake T. Sequential catalysis in organized multienzyme films[M]//Ed. Lvov Y, M?hwald H. Protein architecture: Interfacing molecular assemblies and immobilization biotechnology. New York: Marcel Dekker, Inc., 2000: 169-192.
[14] Lvov Y M. Thin film nanofabrication by alternate adsorption of polyions, nanoparticles, and proteins[M]// Ed. Nalwa R W. Handbook of surfaces and interfaces of materials (Vol. 3). Nanostructured materials, micelles and colloids. San Diego, CA: Academic Press, 2001: 169-188.
[15] Zhou L P, Yang J, Estavillo C, et al. Toxicity screening by electrochemical detection of DNA damage by metabolites generated in situ in ultrathin DNA-enzyme films[J]. Journal of the American Chemical Society, 2003, 125(5): 1431-1436.
[16] Lvov Y M, Lu Z Q, Schenkman J B, et al. Direct electrochemistry of myoglobin and cytochrome P450cam in alternate layer-by-layer films with DNA and other polyions[J]. Journal of the American Chemical Society, 1998, 120(17): 4073-4080
[17] Munge B, Estavillo C, Schenkman J B, et al. Optimization of electrochemical and peroxide-driven oxidation of styrene with ultrathin polyion films containing cytochrome P450cam and myoglobin[J]. ChemBioChem: A European Journal of Chemical Biology, 2003, 4(1): 82-89.
[18] Sultana N, Schenkman J B, Rusling J F. Protein film electrochemistry of microsomes genetically enriched in human cytochrome P450 monooxygenases[J]. Journal of the American Chemical Society, 2005, 127(29): 13460-13461.
[19] Krishnan S, Wasalathanthri D, Zhao L L, et al. Efficient bioelectronic actuation of the natural catalytic pathway of human metabolic cytochrome P450s[J]. Journal of the American Chemical Society, 2011, 133(5): 1459-1465
[20] Huang M H, Xu X, Yang H, et al. Electrochemically-driven and dynamic enhancement of drug metabolism via cytochrome P450 microsomes on colloidal gold/graphene nanocomposites[J]. RSC Advances, 2012, 2(33): 12844-12850.
[21] Sugihara N. Immobilization of cytochrome P450 and electrochemical control of its activity[J]. Polymers for Advanced Technologies, 1998, 9(5): 307-313.
[22] Alonso-Lomilloa M A, Gonzalo-Ruizb J, Domínguez-Renedoa O, et al. CYP450 biosensors based on gold chips for antiepileptic drugs determination[J]. Biosensors & Bioelectronics, 2008, 23(11): 1733-1737.
[23] Dai C, Ding Y, Li M, et al. Direct electrochemistry of cytochrome P450 in a biocompatible film composed of an epoxy polymer and acetylene black[J]. Microchimica Acta, 2012, 176(3/4): 397-404.
[24] Liu S Q, Peng L, Yang X D, et al. Electrochemistry of cytochrome P450 enzyme on nanoparticle-containing membrane-coated electrode and its applications for drug sensing[J]. Analytical Biochemistry, 2008, 375(2): 209-216.
[25] Xu X. Wei W. Huang M H, et al. Electrochemically driven drug metabolism via cytochrome P450 2C9 reductase and indium tin oxide nanoparticle composite[J]. Chemical Communications, 2012, 48(63): 7802-7804.
[26] Sadeghi S J, Fantuzzi A, Gilardi G. Breakthrough in P450 bioelectrochemistry and future perspectives[J]. Biochimica et Biophysica Acta, 2011, 1814(1): 237-24.
[27] Panicco P, Dodhia V R, Fantuzzi A, et al. Enzyme-based amperometric platform to determine the polymorphic response in drug metabolism by cytochromes P450[J]. Analytical Chemistry, 2011, 83(6): 2179-2186.
[28] Fantuzzi A, Fairhead M, Gilardi G. Direct electrochemistry of immobilized human cytochrome P450 2E1[J]. Journal of the American Chemical Society, 2004, 126(16): 5040-5041.
[29] Fantuzzi A, Capria E, Mak L H, et al. An electrochemical microfluidic platform for human P450 drug metabolism profiling[J]. Analytical Chemistry, 2010, 82(24): 10222-10227.
[30] Fantuzzi A, Mak L H, Capria E, et al. A New standardized electrochemical array for drug metabolic profiling with human Cytochromes P450[J]. Analytical Chemistry, 2011, 83(10): 3831-3839.
[31] Tanne J, Schafer D, Khalid W, et al. Light-controlled bioelectrochemical sensor based on CdSe/ZnS quantum dots[J]. Analytical Chemistry, 2011, 83(20): 7778-7785.
[32] Zhao W W, Ma Z Y, Yu P P, et al. Highly sensitive photoelectrochemical immunoassay with enhanced amplification using horseradish peroxidase induced biocatalytic precipitation on a CdS quantum dots multilayer electrode[J]. Analytical Chemistry, 2012, 84(2): 917-923.
[33] Gill R, Zayats M, Willner I. Semiconductor quantum dots for bioanalysis[J]. Angewandte Chemie International Edition, 2008, 47(40): 7602-7625.
[34] Onoda A, Himiyama T, Ohkubo K, et al. Photochemical properties of a myoglobin-CdTe quantum dot conjugate[J]. Chemical Communications, 2012, 48(65): 8054-8056.
[35] Stoll C, Kudera S, Parak W J, et al. Quantum dots on gold: Electrodes for photoswitchable cytochrome c electrochemistry[J]. Small, 2006, 2(6): 741-743l.
[36] Katz E, Zayats M, Willner I, et al. Controlling the direction of photocurrents by means of CdS nanoparticles and cytochromec-mediated biocatalytic cascades[J]. Chemical Communications, 2006, 13: 1395-1397.
[37] Xu X, Qian J, Yu J C, et al. Cytochrome P450 enzyme functionalized-quantum dot as photocatalysts for drug metabolism[J]. Chemical Communications, 2014, 50(57): 7607-7610.



To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.