Recombinant Cytochrome P450 Immobilization for Biosensor Applications

Paternolli, Cristina; Antonini, M.; Ghisellini, Paola; Nicolini, Claudio

doi:10.1021/la048081q

Cited by 30 publications

(20 citation statements)

References 43 publications

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“…In several reports, CYP1A2 peaks were detected at À 380 mV (Krishnan et al, 2011b), À 265 mV (Antonini et al, 2003;Paternolli et al, 2004), À280 mV (Estavillo et al, 2003), and À 437 mV (Shumyantseva et al, 2007). Even more complex is the situation when microsomal CYPs are immobilized on electrodes.…”

Section: Electrochemical Response Of the Mwcnt/mscyp1a2-based Drug Sementioning

confidence: 99%

Continuous monitoring of Naproxen by a cytochrome P450-based electrochemical sensor

Baj-Rossi

Jost²,

Cavallini

et al. 2014

Biosensors and Bioelectronics

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a b s t r a c tThis paper reports the characterization of an electrochemical biosensor for the continuous monitoring of Naproxen based on cytochrome P450. The electrochemical biosensor is based on the drop-casting of multi-walled carbon-nanotubes (MWCNTs) and microsomal cytochrome P4501A2 (msCYP1A2) on a graphite screen-printed electrode (SPE).The proposed biosensor was employed to monitor Naproxen (NAP), a well-known anti-inflammatory compound, through cyclic voltammetry. The dynamic linear range for the amperometric detection of NAP had an upper limit of 300 mM with a corresponding limit of detection (LOD) of 16 7 1 mM (S/N¼3), which is included in NAP physiological range (9-300 mM). The MWCNT/msCYP1A2-SPE sensor was also calibrated for NAP detection in mouse serum that was previously extracted from mice, showing a slightly higher LOD (33718 mM).The stability of the msCYP1A2-based biosensor was assessed by longtime continuous cyclic voltammetric measurements. The ability of the sensor to monitor drug delivery was investigated by using a commercial micro-osmotic pump. Results show that the MWCNT/msCYP1A2-SPE sensor is capable of precisely monitoring the real-time delivery of NAP for 16 h. This work proves that the proposed electrochemical sensor might represent an innovative point-of-care solution for the personalization of drug therapies, as well as for pharmacokinetic studies in both animals and humans.

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Section: Electrochemical Response Of the Mwcnt/mscyp1a2-based Drug Sementioning

confidence: 99%

Continuous monitoring of Naproxen by a cytochrome P450-based electrochemical sensor

Baj-Rossi

Jost²,

Cavallini

et al. 2014

Biosensors and Bioelectronics

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“…Currently, P450 CAM (CYP101), CYP102, CYP1A1, CYP1A2, CYP2B4, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP4A1, CYP11A1, and CYP17A have been used for sensor constructions to monitor drug levels in blood plasma [52, 55-62]. For example, an amperometric drug metabolism biosensor consisting of CYP3A4 encapsulated in a didodecyldimethylammonium bromide vesicular system on a platinum disk electrode was developed for the determination of indinavir, a protease inhibitor antiretroviral drug [55].…”

Section: Applications Of Cytochrome P450 Biocatalystsmentioning

confidence: 99%

Engineering cytochrome P450 biocatalysts for biotechnology, medicine and bioremediation

Kumar

2010

Expert Opinion on Drug Metabolism & Toxicology

149

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Importance of the field: Cytochrome P450 enzymes comprise a superfamily of heme monooxygenases that are of considerable interest for the: 1) synthesis of novel drugs and drug metabolites, 2) targeted cancer gene therapy, 3) biosensor design, and 4) bioremediation. However, their applications are limited because cytochrome P450, especially mammalian P450 enzymes, show a low turnover rate and stability, and require a complex source of electrons through cytochrome P450 reductase and NADPH. Areas covered in this review: In this review, we discuss the recent progress towards the use of P450 enzymes in a variety of above-mentioned applications. We also present alternate and cost-effective ways to perform P450-mediated reaction, especially using peroxides. Furthermore, we expand upon the current progress in P450 engineering approaches describing several recent examples that are utilized to enhance heterologous expression, stability, catalytic efficiency, and utilization of alternate oxidants. What the reader will gain: The review will provide a comprehensive knowledge in the design of P450 biocatalysts for potentially practical purposes. Finally, we provide a prospective on the future aspects of P450 engineering and its applications in biotechnology, medicine, and bioremediation. Take home message: Because of its wide applications, academic and pharmaceutical researchers, environmental scientists, and health care providers are expected to gain current knowledge and future prospects of the practical use of P450 biocatalysts.

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“…Traditionally, P450 monooxygenases have been applied in drug toxicity testing to predict the in vivo human metabolism and effects of prodrugs and other xenobiotics (Miners, 2000; Guengerich 2002; Ingelman-Sundberg 2004; Guengerich 2006). Considering that these enzymes catalyze diverse reactions in regio- and stereo-selective manner, their properties have been investigated for various pharmaceutical, biotechnological, and environmental applications such as in drug discovery and development, production of fine chemicals, fragrances, pharmaceutical compounds and biofuels, biosensing, and bioremediation (Guengerich 1995, 2002 & 2006; Ingelman-Sundberg 2004; Urlacher and Eiben 2006; Paternolli et al 2004; Zhang et al 2011). These P450 applications have been thoroughly reviewed elsewhere (Guengerich 2002; Urlacher and Eiben 2006; Zhang et al 2011).…”

Section: Introductionmentioning

confidence: 99%

P450 monooxygenases (P450ome) of the model white rot fungusPhanerochaete chrysosporium

Syed

Yadav

2012

Critical Reviews in Microbiology

118

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Phanerochaete chrysosporium, the model white rot fungus, has been the focus of research for the past about four decades for understanding the mechanisms and processes of biodegradation of the natural aromatic polymer lignin and a broad range of environmental toxic chemicals. The ability to degrade this vast array of xenobiotic compounds was originally attributed to its lignin-degrading enzyme system (LDS), mainly the extracellular peroxidases. However, subsequent physiological, biochemical, and/or genetic studies by us and others identified the involvement of a peroxidase-independent oxidoreductase system, the cytochrome P450 monooxygenase system. The whole genome sequence revealed an extraordinarily large P450 contingent (P450ome) with an estimated 149 P450s in this organism. This review focuses on the current status of understanding on the P450 monooxygenase system of P. chrysosporium in terms of pre-genomic and post-genomic identification, structural and evolutionary analysis, transcriptional regulation, redox partners, and functional characterization for its biodegradative potential. Future research on this catalytically diverse oxidoreductase enzyme system and its major role as a newly emerged player in xenobiotic metabolism/degradation is discussed.

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Recombinant Cytochrome P450 Immobilization for Biosensor Applications

Cited by 30 publications

References 43 publications

Continuous monitoring of Naproxen by a cytochrome P450-based electrochemical sensor

Continuous monitoring of Naproxen by a cytochrome P450-based electrochemical sensor

Engineering cytochrome P450 biocatalysts for biotechnology, medicine and bioremediation

P450 monooxygenases (P450ome) of the model white rot fungusPhanerochaete chrysosporium

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