Colorimetric detection of immobilized horseradish peroxidase based on the co-oxidation of benzidine derivatives and 4-chloro-1-naphthol

Ulyashova, M.M.; Rubtsova, M. Yu.; Egorov, A. M.

doi:10.1007/s11172-011-0101-3

Cited by 8 publications

(6 citation statements)

References 16 publications

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“…2, see SI for details). 34 Indeed, the nonequilibrium microphases of SH showed rapid peroxidase activity with vi =2.35 ± 0.52 µM/min. Further, the consumption of CN with time was monitored via HPLC chromatograms (Fig.…”

Section: Resultsmentioning

confidence: 96%

“…Hence, the selection of the substrate was based on two criteria, firstly, it should be a substrate for extant peroxidase enzymes and secondly, it should be aromatic as planar moieties are known to induce assembly of fatty acid based amphiphiles. 33,34 Based on these, planar aromatic molecule 4-chloro-1naphthol (CN) was selected (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

“…1). 34 Indeed, addition of CN (14 mM) resulted in the rapid selfassembly of SH (12 mM) in presence of hemin (0.15 mM) as within ca. 30 min the solution turned into a self-supporting gel (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Emergence of a Promiscuous Peroxidase Under Non-Equilibrium Conditions

Pal¹,

Reja²,

Bal³

et al. 2021

Preprint

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In the far from equilibrium state of living matter, energy is consumed to fuel the metabolic networks of enzymatic reactions. The emergence of protometabolic pathways in primeval earth is intricately related to the evolutionary journey of modern enzymes. Fundamental understanding of such energy driven generation of early catalytic systems would help us recognize the conditions required for the minimal metabolism that predated the chemical emergence of life. Herein, we report substrate driven generation of a non-equilibrium catalytic machinery from a single amino acid functionalized fatty acid in presence of a cofactor hemin. The non-equilibrium assembled state showed acceleration of catalytic potential resulting in degradation of the substrate and subsequently led to disassembly. Controls that promoted equilibration could not access the three-dimensional microphases and showed substantially lower catalytic activity. Significantly, the assembled state showed latent catalytic function by hydrolysing a precursor to yield the same substrate. Consequently, the assembly was benefitted with augmented lifetime of the catalytic state exploiting a promiscuous cascade and thus foreshadowing protometabolism. The results contribute towards our understanding of energy driven generation of primitive catalytic machineries that assisted the minimal metabolism of early life.

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Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Emergence of a Promiscuous Peroxidase Under Non-Equilibrium Conditions

Pal¹,

Reja²,

Bal³

et al. 2021

Preprint

View full text Add to dashboard Cite

show abstract

“…Our method is characterized by sensitivity, comparable to the best analytical performance of other methods, while it should be noted that the longest DNA fragment was determined in our study. When compared to hybridization analysis of the same genes based on the same capture probes on DNA microchips with colorimetric detection, the LOD of our method is more than three orders of magnitude lower; also, the dynamic range is 1-2 orders of magnitude wider [42]. Comparing the analytical characteristics of the determination of short and long DNA fragments, it can be assumed that the lengthening of the capture probes may contribute to an increase in the sensitivity of the method.…”

Section: Digital Quantification Of Nucleic Acids On Silicon Microchipsmentioning

confidence: 93%

Multiplex Digital Quantification of β-Lactamase Genes in Antibiotic-Resistant Bacteria by Counting Gold Nanoparticle Labels on Silicon Microchips

et al. 2022

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Digital quantification based on counting of individual molecules is a promising approach for different biomedical applications due to its enhanced sensitivity. Here, we present a method for the digital detection of nucleic acids (DNA and RNA) on silicon microchips based on the counting of gold nanoparticles (GNPs) in DNA duplexes by scanning electron microscopy (SEM). Biotin-labeled DNA is hybridized with capture oligonucleotide probes immobilized on the microchips. Then biotin is revealed by a streptavidin–GNP conjugate followed by the detection of GNPs. Sharp images of each nanoparticle allow the visualization of hybridization results on a single-molecule level. The technique was shown to provide highly sensitive quantification of both short oligonucleotide and long double-strand DNA sequences up to 800 bp. The lowest limit of detection of 0.04 pM was determined for short 19-mer oligonucleotide. The method’s applicability was demonstrated for the multiplex quantification of several β-lactamase genes responsible for the development of bacterial resistance against β-lactam antibiotics. Determination of nucleic acids is effective for both specific DNA in lysates and mRNA in transcripts. The method is also characterized by high selectivity for single-nucleotide polymorphism discrimination. The proposed principle of digital quantification is a perspective for studying the mechanisms of bacterial antibiotic resistance and bacterial response to drugs.

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“…Consequently, we probed whether the generated nanostructures could recruit other molecules that in combination would be able to impart a latent, i.e., promiscuous, catalytic role to degrade the aromatic alcohol, a substrate of peroxidases. 43 With this idea, the cofactor of natural peroxidases, i.e., hemin (H) was introduced in the system to perform the oxidation step. Interestingly, upon addition of H (0.…”

mentioning

confidence: 99%

Temporal (Dis)Assembly of Peptide Nanostructures Dictated by Native Multistep Catalytic Transformations

Pal,

Saha,

Das

2024

Nano Lett.

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Emergence of complex catalytic machinery via simple building blocks under non-equilibrium conditions can contribute toward the system level understanding of the extant biocatalytic reaction network that fuels metabolism. Herein, we report temporal (dis)assembly of peptide nanostructures in presence of a cofactor dictated by native multistep cascade transformations. The short peptide can form a dynamic covalent bond with the thermodynamically activated substrate and recruit cofactor hemin to access non-equilibrium catalytic nanostructures (positive feedback). The neighboring imidazole and hemin moieties in the assembled state rapidly converted the substrate to product(s) via a two-step cascade reaction (hydrolase-peroxidase like) that subsequently triggered the disassembly of the catalytic nanostructures (negative feedback). The feedback coupled reaction cycle involving intrinsic catalytic prowess of short peptides to realize the advanced trait of two-stage cascade degradation of a thermodynamically activated substrate foreshadows the complex non-equilibrium protometabolic networks that might have preceded the chemical emergence of life.

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Colorimetric detection of immobilized horseradish peroxidase based on the co-oxidation of benzidine derivatives and 4-chloro-1-naphthol

Cited by 8 publications

References 16 publications

Emergence of a Promiscuous Peroxidase Under Non-Equilibrium Conditions

Emergence of a Promiscuous Peroxidase Under Non-Equilibrium Conditions

Multiplex Digital Quantification of β-Lactamase Genes in Antibiotic-Resistant Bacteria by Counting Gold Nanoparticle Labels on Silicon Microchips

Temporal (Dis)Assembly of Peptide Nanostructures Dictated by Native Multistep Catalytic Transformations

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