Abstract
l-Amino acid oxidases (LAO) are widely distributed enzymes but those from snake venoms have been studied the most. We describe a method for in-gel detection of LAO activities based on H2O2 detection by a horseradish peroxidase-coupled reaction using o-phenylenediamine. Complex substrates and single l-amino acids were used successfully for screening LAO activities in higher fungi using crude aqueous extracts of fruiting bodies of 22 basidiomycetes and 1 ascomycete. Half of these samples exhibited one to two bands of LAO activities with mostly broad substrate specificities and a variety of apparent molecular masses ranging from 25 to 200 kDa that were generally more active at pH 5.5 than at pH 8.0. Mushrooms are shown to be a rich source of LAOs that could find use in various medical and biotechnological applications.
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Abbreviations
- CSM:
-
Complete supplement mixture
- FAD:
-
Flavin adenine dinucleotide
- HRP:
-
Horseradish peroxidase
- LAO:
-
l-Amino acid oxidase
- OPD:
-
o-Phenylenediamine
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Acknowledgements
We are grateful to Dr. Adrijana Leonardi for a sample of horned viper venom, to Dr. Jože Brzin for collecting mushroom samples, and to Prof. Roger H. Pain for critical reading of the manuscript and language editing. This work was supported by the Slovenian Research Agency (Grant Number P4-0127 to J.K.).
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Žun, G., Kos, J. & Sabotič, J. Higher fungi are a rich source of l-amino acid oxidases. 3 Biotech 7, 230 (2017). https://doi.org/10.1007/s13205-017-0813-7
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DOI: https://doi.org/10.1007/s13205-017-0813-7