Roughened graphite biointerfaced with P450 liver microsomes: Surface and electrochemical characterizations

Walker, Austin; Walgama, Charuksha; Nerimetla, Rajasekhara; Alavi, S. Habib; Echeverría, Elena; Harimkar, Sandip P.; McIlroy, David N.; Krishnan, Sadagopan

doi:10.1016/j.colsurfb.2020.110790

Cited by 16 publications

(8 citation statements)

References 17 publications

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“…Figure A represents the cyclic voltammograms of as acquired, and the background subtracted human liver microsomal biofilm adsorbed onto the amine-enriched pyrenyl-nanocarbon-modified HPG electrodes. The formal potential observed around −0.45 V vs Ag/AgCl (pH 7.0, 0.3 V s –1 scan rate) was consistent with prior reports on liver microsomal and P450 bactosomal biofilms on carbon and gold-self-assembled monolayer electrodes, which was identified to originate from the membrane-bound microsomal P450-reductase as the electron receiver from the electrode. ,, …”

Section: Resultssupporting

confidence: 89%

“…While biochemical in vitro assays and methods are valuable, an alternate complementing analytical technique is to drive the purified P450 and microsomal biocatalysis electrochemically under an applied reductive potential with simplicity, creating a bioelectrocatalytic system. , In this system, the P450 enzyme is attached to an electrode surface either in the purified form or bound to the microsomal membrane or other support materials. − When a substrate is added to the system, the P450 enzyme catalyzes the oxidation of the substrate, generating an electrochemical signal that can be measured. In the same system, in the place of substrates, specific P450 inhibitors can be used to profile the activity of a specific P450 isoform toward a drug conversion into metabolites …”

Section: Introductionmentioning

confidence: 99%

“…The electrochemical driving of immobilized microsomal P450 electrocatalysis is a cost-effective, efficient probing of the enzymes and their reductase-mediated electron transfer pathway for the direct measurement of P450 activity and drug metabolism and inhibition assays. ,, Moreover, the recent advancements allowing the successful direct use of human liver microsomes and bacterial-expressed-specific P450-isoform called bactosomes for bioelectrocatalysis have revolutionized the field significantly. ,, By immobilizing the P450 enzyme onto an electrode surface, we can also create a highly sensitive and selective biosensor for the detection of drugs, environmental pollutants, and other xenobiotics. ,− …”

Section: Introductionmentioning

confidence: 99%

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Nanobioelectrocatalysis Using Human Liver Microsomes and Cytochrome P450 Bactosomes: Pyrenyl-Nanocarbon Electrodes

Premaratne,

Niroula,

Moulton

et al. 2024

ACS Appl. Bio Mater.

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Human liver microsomes containing various drugmetabolizing cytochrome P450 (P450) enzymes, along with their NADPH-reductase bound to phospholipid membranes, were absorbed onto 1-pyrene butylamine pi−pi stacked with aminefunctionalized multiwalled carbon nanotube-modified graphite electrodes. The interfaced microsomal biofilm demonstrated direct electrochemical communication with the underlying electrode surface and enhanced oxygen reduction electrocatalytic activity typical of heme enzymes such as P450s over the unmodified electrodes and nonenzymatic currents. Similar enhancements in currents were observed when the bioelectrodes were constructed with recombinant P450 2C9 (single isoform) expressed bactosomes. The designed liver microsomal and 2C9 bactosomal bioelectrodes successfully facilitated the electrocatalytic conversion of diclofenac, a drug candidate, into 4′-hydroxydiclofenac. The enzymatic electrocatalytic metabolite yield was several-fold greater on the modified electrodes than on the unmodified bulk graphite electrodes adsorbed with a microsomal or bactosomal film. The nonenzymatic metabolite production was less than the enzymatically catalyzed metabolite yield in the designed microsomal and bactosomal biofilm electrodes. To test the throughput potential of the designed biofilms, eight-electrode array configurations were tested with the microsomal and bactosomal biofilms toward electrochemical 4′-hydroxydiclofenac metabolite production from diclofenac. The stability of the designed microsomal bioelectrode was assessed using nonfaradaic impedance spectroscopy over 40 h, which indicated good stability.

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Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Nanobioelectrocatalysis Using Human Liver Microsomes and Cytochrome P450 Bactosomes: Pyrenyl-Nanocarbon Electrodes

Premaratne,

Niroula,

Moulton

et al. 2024

ACS Appl. Bio Mater.

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“…Microbial-Inspired Surface Patterning. The graphite sheet (15 mm thick) was first subjected to various grades of M-SiC papers (60 to 1500) using a DST610 belt sander to achieve various degrees of surface roughness 25 and then degreased in acetone (99.5%) and ethanol (99.9%) serially as per the standard protocols. 26 Then, the graphite sheet was cleaned with deionized water, followed by ultrasonication and incubated in 10% H 2 SO 4 and LB medium for 24 h. The pre-treated graphite sheet was then sterilized by UV irradiation and stored aseptically until use.…”

Section: Bacterial Culture Conditionsmentioning

confidence: 99%

Microbial-Inspired Surface Patterning for Selective Bacterial Actions for Enhanced Performance in Microbial Fuel Cells

Bijimol

Sreelekshmy

Kumar

et al. 2022

ACS Appl. Bio Mater.

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The performance of any bio-electrochemical system is dependent on the efficiency of electrode−microbial interactions. Surface properties play a focal role in bacterial attachment and biofilm formation on the electrodes. In addition to electrode surface properties, selective bacterial adhesion onto the electrode surface is mandatory to mitigate energy loss due to undesired bacterial interactions on the electrode surface. In the present study, microbial-patterned graphite scaffolds are developed for selective bacterial−electrode interactions. A power density as high as 1105 mW/m 2 is achieved with mG-E (a graphite electrode patterned with Escherichia coli), which is about 3 times higher than that of the pristine graphite electrode (370 mW/m 2 ). Initial mechanical pre-treatment of the graphite electrode, followed by bacterial patterning, results in the formation of a unique cobblestone topography with a tuned surface area of 127.12 m 2 /g. This provides suitable morphology with enhanced active sites for selective bacterial intercalation in graphite layers. This cannot be otherwise achieved by any mechanical or other means. A unique methodology of symbolic regression is adopted to validate a genetic algorithm suitable for predicting a perfect correlation between surface characteristics and electrochemical characteristics with a minimum root-mean-square error of 0.08. The bacterial intercalation onto the graphite electrode causes protuberance of the graphite layers that reduces the surface potential and resistance, leading to high electron transfer. The study presents a unique bacterial-inspired surface patterning on the anode, which is critical for the performance of a microbial fuel cell.

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“…Improvements in cytochrome P450 electrocatalysis remain a significant challenge. Various approaches have been proposed for the creation of electrochemical systems that retain and increase the catalytic activity of the enzymes [ 6 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 ]. Moreover, enzyme immobilization in a volume-confined environment could increase its stability and reduce denaturation and protein unfolding [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs

et al. 2023

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In the present work, screen-printed electrodes (SPE) modified with a synthetic surfactant, didodecyldimethylammonium bromide (DDAB) and streptolysin O (SLO) were prepared for cytochrome P450 3A4 (CYP3A4) immobilization, direct non-catalytic and catalytic electrochemistry. The immobilized CYP3A4 demonstrated a pair of redox peaks with a formal potential of −0.325 ± 0.024 V (vs. the Ag/AgCl reference electrode). The electron transfer process showed a surface-controlled mechanism (“protein film voltammetry”) with an electron transfer rate constant (ks) of 0.203 ± 0.038 s−1. Electrochemical CYP3A4-mediated reaction of N-demethylation of erythromycin was explored with the following parameters: an applied potential of −0.5 V and a duration time of 20 min. The system with DDAB/SLO as the electrode modifier showed conversion of erythromycin with an efficiency higher than the electrode modified with DDAB only. Confining CYP3A4 inside the protein frame of SLO accelerated the enzymatic reaction. The increases in product formation in the reaction of the electrochemical N-demethylation of erythromycin for SPE/DDAB/CYP3A4 and SPE/DDAB/SLO/CYP3A4 were equal to 100 ± 22% and 297 ± 7%, respectively. As revealed by AFM images, the SPE/DDAB/SLO possessed a more developed surface with protein cavities in comparison with SPE/DDAB for the effective immobilization of the CYP3A4 enzyme.

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Roughened graphite biointerfaced with P450 liver microsomes: Surface and electrochemical characterizations

Cited by 16 publications

References 17 publications

Nanobioelectrocatalysis Using Human Liver Microsomes and Cytochrome P450 Bactosomes: Pyrenyl-Nanocarbon Electrodes

Nanobioelectrocatalysis Using Human Liver Microsomes and Cytochrome P450 Bactosomes: Pyrenyl-Nanocarbon Electrodes

Microbial-Inspired Surface Patterning for Selective Bacterial Actions for Enhanced Performance in Microbial Fuel Cells

Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs

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