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Extracellular Oxidase from the Neonothopanus nambi Fungus as a Promising Enzyme for Analytical Applications

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Abstract

The extracellular enzyme with oxidase function was extracted from the Neonothopanus nambi luminescent fungus by using mild processing of mycelium with β-glucosidase and then isolated by gel-filtration chromatography. The extracted enzyme is found to be a FAD-containing protein, catalyzing phenol co-oxidation with 4-aminoantipyrine without addition of H2O2, which distinguishes it from peroxidases. This fact allowed us to assume that this enzyme may be a mixed-function oxidase. According to gel-filtration chromatography and SDS-PAGE, the oxidase has molecular weight of 60 kDa. The enzyme exhibits maximum activity at 55–70 °C and pH 5.0. Kinetic parameters Km and Vmax of the oxidase for phenol were 0.21 mM and 0.40 µM min−1. We suggest that the extracted enzyme can be useful to develop a simplified biosensor for colorimetric detection of phenol in aqueous media, which does not require using hydrogen peroxide.

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Availability of data and materials

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

4-AAP:

4-Aminoantipyrine (1-phenyl-2,3-dimethyl-4-aminopyrazolone)

BSA:

Bovine serum albumin

DI:

Deionized water

FAD:

Flavin adenine dinucleotide

SDS-PAGE:

Sodium dodecyl sulfate- polyacrylamide gel electrophoresis

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Funding

This work was supported by the state budget allocated to the fundamental research at the Russian Academy of Sciences, Project No. 0356–2019-0022.

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Authors and Affiliations

  1. Institute of Biophysics, Siberian Branch of Russian Academy of Sciences, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, Russia, 660036

    Olga Mogilnaya, Nikita Ronzhin, Ekaterina Posokhina & Vladimir Bondar

Authors
  1. Olga Mogilnaya
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  2. Nikita Ronzhin
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  3. Ekaterina Posokhina
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  4. Vladimir Bondar
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All authors made substantial contributions in conceptualizing, drafting, developing and reviewing the manuscript. The paper was reviewed and approved by all authors prior to submission for peer review.

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Correspondence to Olga Mogilnaya.

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Mogilnaya, O., Ronzhin, N., Posokhina, E. et al. Extracellular Oxidase from the Neonothopanus nambi Fungus as a Promising Enzyme for Analytical Applications. Protein J 40, 731–740 (2021). https://doi.org/10.1007/s10930-021-10010-z

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