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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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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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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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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DOI: https://doi.org/10.1007/s10930-021-10010-z