Electrocatalysis by subcellular liver fractions bound to carbon nanostructures for stereoselective green drug metabolite synthesis

Nerimetla, Rajasekhara; Krishnan, Sadagopan

doi:10.1039/c5cc03364k

Cited by 24 publications

(13 citation statements)

References 28 publications

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“…22,34 Furthermore, the conjugates prepared with 10 mg of MNPs produced larger amounts of product after five uses compared to the conjugates made using 1 and 5 mg of MNPs (Fig. 6).…”

Section: Resultsmentioning

confidence: 91%

Stability, scalability, and reusability of a volume efficient biocatalytic system constructed on magnetic nanoparticles

Premaratne

Nerimetla

Matlock

et al. 2016

Catal. Sci. Technol.

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This report investigates for the first time stability, scalability, and reusability characteristics of a protein nano-bioreactor useful for green synthesis of fine chemicals in aqueous medium extracting maximum enzyme efficiency. Enzyme catalysts conjugated with magnetic nanomaterials allow easy product isolation after a reaction involving simple application of a magnetic field. In this study, we examined a biocatalytic system made of peroxidase-like myoglobin (Mb), as a model protein, to covalently conjugate with poly(acrylic acid) functionalized magnetic nanoparticles (MNPs, 100 nm hydrodynamic diameter) to examine the catalytic stability, scalability, and reusability features of this bioconjugate. Application of the conjugate was effective for electrochemical reduction of organic and inorganic peroxides, and for both peroxide-mediated and electrocatalytic oxidation of the protein substrate 2, 2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with greater turnover rates and product yields than Mb prepared in solution or MNP alone. Mb-attached MNPs displayed extensive catalytic stability even after 4 months of storage compared to Mb present in solution. Five- and ten-fold scale up of MNPs in the bioconjugates resulted in two- and four-fold increases in protein-catalyzed oxidation products, respectively. Nearly 40% of the initial product was present even after four reuses, which is advantageous for synthesizing sufficient products with a minimal investment of precious enzymes. Thus, the results obtained in this study are highly significant in guiding cost-effective development and efficient multiple uses of enzyme catalysts for biocatalytic, electrocatalytic, and biosensing applications via magnetic nanomaterials conjugation.

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“…22,34 Furthermore, the conjugates prepared with 10 mg of MNPs produced larger amounts of product after five uses compared to the conjugates made using 1 and 5 mg of MNPs (Fig. 6).…”

Section: Resultsmentioning

confidence: 91%

Stability, scalability, and reusability of a volume efficient biocatalytic system constructed on magnetic nanoparticles

Premaratne

Nerimetla

Matlock

et al. 2016

Catal. Sci. Technol.

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“…86 They note that residual product yield after four cycles of 1 h each for P450-HLM-MWCNTs was down to 10%, confirming the challenge of designing a reusable electrocatalytic P450 film. 86 Recently, the same group applied magnetic nanoparticles (MNPs) to immobilize HLM for metabolite and drug biosensing to increase enzyme loading and to obtain easier separation from the media compared with MWCNTs. 87 MNPs have received significant attention in enzymatic electrocatalysis and as carriers for heterogeneous biocatalysis.…”

Section: P450smentioning

confidence: 94%

“…102 PDA increased sensitivity (in μA M −1 ) 2.3-fold, with K M app (testosterone) in the same range (1.6-fold lower) and twice the half-life of HLM adsorbed onto MWCNTs coated on pyrolytic graphite electrodes. 86 The coated PDA film provided a promising platform for facile immobilization of HLM to design a biosensor with good electrode-driven catalytic performance and long half-life. 102 Biocatalytic Applications Using Covalently Immobilized P450 BM3.…”

Section: Weber Et Al Studied Adsorption Of the Heme Domain Of P450mentioning

confidence: 99%

“…Nerimetla and Krishnan reported that adsorption of P450-containing human liver microsomes (P450-HLM) on MWCNTs resulted in ∼1.7-fold lower apparent Michaelis constant ( K M app ) and ∼5-fold higher metabolite yield for testosterone hydroxylation compared to unmodified electrodes (Table ). They note that residual product yield after four cycles of 1 h each for P450-HLM-MWCNTs was down to 10%, confirming the challenge of designing a reusable electrocatalytic P450 film …”

Section: Immobilization Of P450s By Physical Adsorption On Pre-existi...mentioning

confidence: 99%

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An Overview of Cytochrome P450 Immobilization Strategies for Drug Metabolism Studies, Biosensing, and Biocatalytic Applications: Challenges and Opportunities

2021

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Cytochrome P450s (P450s or CYPs) are a large superfamily of ubiquitous heme proteins. Consistent with their natural roles in oxidative metabolism of a broad range of substrates including antibiotics and xenobiotics and in the biosynthesis of complex natural products, there are two main biotechnological applications: drug metabolism studies and biocatalytic transformations. Metabolic analyses are typically performed with the P450 enzyme immobilized on a biosensing platform. Biocatalytic reactions also increasingly make use of immobilization methods with the goal of improving operational stability of the P450s. In this Review, we briefly introduce the growing field of enzyme immobilization then focus on the advances, benefits, and challenges that apply specifically to P450s. Classical immobilization methods are presented, and recently developed materials and strategies that address the need for operational stability, the multicomponent structure of P450s, their requirement for a source of electrons, and the need to overcome limited O 2 availability are discussed in the context of metabolic, biosensing, and biocatalytic applications.

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“…Additionally, our current research focuses on understanding the electrocatalytic properties of human liver microsomes on various electrodes for high throughput drug metabolism and inhibition assays. 5,6 © The Electrochemical Society. All rights reserved.…”

Section: (B) (A) (B) (C) (D) (C)mentioning

confidence: 99%

The 2016 Joseph W. Richards Summer Research Fellowship — Summary Report: Single Drop Electroanalysis for Low Cost Quality Control Testing of Oxidative Pharmaceuticals

Walgama

2016

Electrochem. Soc. Interface

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Q uality control analysis of biochemically relevant pharmaceuticals is an important process in the drug industry. The therapeutic value of a drug mainly depends on the purity of the active pharmaceutical ingredient (API). Various chromatographic, spectroscopic, titrimetric, and electrochemical methods have been used in pharmaceutical industry for the estimation of APIs in drugs.1 Electroanalytical methods can be tailored to provide unique advantages such as high sensitivity, low cost, portability, fast assay time, and automation features. These are attractive for industrial-scale analytical applications. Fig. 1. (A) Schematic representation of single drop analysis using a MWCNT-COOH modified SPE (WE) with a carbon counter electrode (CE) and a silver pseudo-reference electrode (RE). (B) CVs (i) at lower concentration range and (ii) at higher concentration range. (C) Oxidation peak currents versus concentration of 50 µL droplets of varying concentrations of analytes placed on2 Moreover, measurements in micro-volume droplets of pharmaceutically important compounds allow miniaturized design, ease of operation, opportunities for throughput analysis, and less sample consumption.3 In this study, we measured oxidative pharmaceuticals as single 50 µL drops on screen printed electrodes (SPEs) modified with carboxylated multiwalled carbon nanotubes (MWCNT-COOH). Acetaminophen (a pain-killer), ascorbic acid (a dietary supplement), nicotinamide adenine dinucleotide reduced form (NADH) (a dietary supplement), and Nicotine (the active agent of antismoking pharmaceuticals) were analyzed. The relationship between polar/nonpolar interactions of analytes and surface carbon nanostructures in influencing sensitivity and limit of detection was understood.Cyclic voltammograms (CVs) of various concentrations of the four analytes prepared in pH 7.0 phosphate buffer were obtained on SPEs, [110 CNT, carbon counter electrode and silver pseudo-reference electrode, Dropsens Ltd.,Llanera (Asturias) Spain] upon which 50 µL drops were added (Fig.

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Electrocatalysis by subcellular liver fractions bound to carbon nanostructures for stereoselective green drug metabolite synthesis

Cited by 24 publications

References 28 publications

Stability, scalability, and reusability of a volume efficient biocatalytic system constructed on magnetic nanoparticles

Stability, scalability, and reusability of a volume efficient biocatalytic system constructed on magnetic nanoparticles

An Overview of Cytochrome P450 Immobilization Strategies for Drug Metabolism Studies, Biosensing, and Biocatalytic Applications: Challenges and Opportunities

The 2016 Joseph W. Richards Summer Research Fellowship — Summary Report: Single Drop Electroanalysis for Low Cost Quality Control Testing of Oxidative Pharmaceuticals

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