Laccase as a New Enzymatic Label for Enzyme Immunoassay

Skorobogat'ko, O. V.; Gindilis, A. L.; Troitskaya, Elena N.; Shuster, Alexander M.; Yaropolov, A. I.

doi:10.1080/00032719408000307

Cited by 13 publications

(6 citation statements)

References 11 publications

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“…Moreover, laccase has been suggested as a useful agent for treating contaminated water, and removing phenol constituents in various food or beverages . In academic research, laccase has been used for the chemical transformations of small molecules, production of polymers, as an enzyme in ELISA, and in biosensing applications . This research has led to the development of modified laccases with a variety of properties, including stability in organic solvents and improved reactivity in biological media …”

Section: Figurementioning

confidence: 99%

Laccase‐Mediated Catalyzed Fluorescent Reporter Deposition for Live‐Cell Imaging

Cisneros

Devaraj

2019

ChemBioChem

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Catalyzed reporter deposition (CARD) is a widely established method for labeling biological samples analyzed using microscopy. Horseradish peroxidase, commonly used in CARD to amplify reporter signals, requires the addition of hydrogen peroxide, which may perturb samples used in live‐cell microscopy. Herein we describe an alternative method of performing CARD using a laccase enzyme, which does not require exogenous hydrogen peroxide. Laccase is an oxidative enzyme which can carry out single‐electron oxidations of phenols and related compounds by reducing molecular oxygen. We demonstrate proof‐of‐concept for this technique through the nontargeted covalent labeling of bovine serum albumin using a fluorescently labeled ferulic acid derivative as the laccase reporter substrate. We further demonstrate the viability of this approach by performing live‐cell CARD with an antibody‐conjugated laccase against a surface‐bound target. CARD using laccase produces an amplified fluorescence signal by labeling cells without the need for exogenous hydrogen peroxide.

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Section: Figurementioning

confidence: 99%

Laccase‐Mediated Catalyzed Fluorescent Reporter Deposition for Live‐Cell Imaging

Cisneros

Devaraj

2019

ChemBioChem

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“…Laccase catalysis could be useful as biosensors for detecting O 2 and wide verity of reducing substrates. Studies suggest that laccase can also be used for enzyme immunoassay (Skorobogat'ko et al 1994).…”

Section: Introductionmentioning

confidence: 99%

Physicochemical characterization, structural analysis and homology modeling of bacterial and fungal laccases using in silico methods

Tamboli

Rane

Patil

et al. 2015

Netw Model Anal Health Inform Bioinforma

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Laccases from various sources like higher plants, fungi as well as bacteria are known to have wide range of applications in various industries such as paper, textile and beverage. This enzyme is extensively studied because of its vital role in bioremediation, biosensors and diagnostics. Phylogenetic analysis of retrieved sequences of laccases from selected bacteria and fungi inferred variation in sequences and these laccases were grouped independently. All laccases of selected organisms are acidic showing computed pI \ 7 and hydrophilic in nature. Presence of disulfide bridges was observed in laccases of bacteria Pseudomonas putida, Haemophilus parasuis HPS11 and Endozoicomonas numazuensis as well as in all selected fungal laccases. N-myristoylation and phosphorylation sites for Protein kinase C and Casein kinase II were found in all selected laccases. Secondary structure prediction revealed dominance of random coils in selected laccases. Validation results of predicted 3D structures of laccase enzymes confirmed that the modeled structures were of good quality. All selected bacterial laccases were intracellular and those of fungal were found to be extracellular. These investigations provide functional information about these laccases and may help to design successful experimental work aiming towards isolation and purification of laccases from closely related organisms. Experimental structures of these laccase enzymes are not yet available, so till then this study will provide a platform for knowing the structural organization responsible for functioning of these enzymes.

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“…[6] Moreover,l accase has been suggested as au seful agent for treating contaminated water, [7] and removingp henol constituents in various food or beverages. [8] In academic research,l accase has been used for the chemicalt ransformationso fs mall molecules, [9] production of polymers, [10] as an enzymei nE LISA, [11][12][13] and in biosensing applications. [14] This research has led to the development of modified laccases with av ariety of properties,i ncluding stability in organic solvents [15] and improved reactivity in biological media.…”

mentioning

confidence: 99%

Laccase-mediated catalyzed fluorescent reporter deposition for live cell imaging

Cisneros¹,

Devaraj²

2019

Preprint

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Catalyzed reporter deposition (CARD) is a widely established method for labeling biological samples analyzed using microscopy. Horseradish peroxidase, commonly used in CARD to amplify reporter signals, requires the addition of hydrogen peroxide which may perturb samples used in live-cell microscopy. Herein we describe an alternative method of performing CARD using a laccase enzyme, which does not require exogenous hydrogen peroxide. Laccase is an oxidative enzyme which can carry out single-electron oxidations of phenols and related compounds by reducing molecular oxygen. We demonstrate proof-of-concept for this technique through the non-targeted covalent labeling of bovine serum albumin using a fluorescently-labeled ferulic acid derivative as the laccase reporter substrate. We further demonstrate the viability of this approach by performing live-cell CARD with an antibody-conjugated laccase against a surface bound target. CARD using laccase produces an amplified fluorescence signal by labeling cells without the need for exogenous hydrogen peroxide.

show abstract

Laccase as a New Enzymatic Label for Enzyme Immunoassay

Cited by 13 publications

References 11 publications

Laccase‐Mediated Catalyzed Fluorescent Reporter Deposition for Live‐Cell Imaging

Laccase‐Mediated Catalyzed Fluorescent Reporter Deposition for Live‐Cell Imaging

Physicochemical characterization, structural analysis and homology modeling of bacterial and fungal laccases using in silico methods

Laccase-mediated catalyzed fluorescent reporter deposition for live cell imaging

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