Modification of isolation methods and physico-biochemical properties of preparations of fungal oxidoreductases

Keywords: Basidiomycetes; catalases; peroxidases; secretions; enzymatic preparations; characteristics


The results of the modification of methods for producing enzymatic preparations (EP) of peroxidases and catalases of extra- and intracellular finding from fungal cultures are presented. Strains of Flammulina velutipes F-vv, Lentinula edodes 523 and Pleurotus ostreatus P-01 were used as producers of oxidoreductases. The producers were cultured in glucose-peptone medium, modified for each strain. Protein fractionation was carried out by leaching with ammonium sulfate at a saturation of 40–70% for peroxidases and 80% for catalases. The obtained solutions of protein fractions were additionally subjected to purification by dialysis, gel filtration on Molselect G-50 and G-75 granules, and also freeze drying. The yield of enzymatic preparations per unit mass of mycelium and the volume of culture fluid were calculated. The individual characteristics of EP – enzymatic activity, the mass percentage of protein and associated amino acids, the ratio of the latter in groups depending on the nature of the radicals (amphotericity) of protein molecules are established. It was proved that the amino acid content in the proteins of fungal EP catalases and peroxidases indicates their acidic nature and this is confirmed by the pH values of aqueous solutions. Examination and toxicity testing of enzymatic preparations were carried out in certified laboratories, which confirmed their characteristics and compliance with safety requirements. The therapeutic properties of amino acids that are part of proteins or are in a free state in enzyme preparations are analyzed. In this way, the methods have been developed for producing enzymatic preparations of peroxidases and catalases of extra- and intracellular location, which allow new antioxidant enzymes with individual properties to be obtained, and, as a result, bring prospects for use in various industries and scientific research.


Bisko, N. A., Lomberg, M. L., Mytropolska, N. Y., & Mykchaylova, O. B. (2016). The IBK mushroom culture collection. Kyiv, M. G. Kholodny Institute of Botany, National Academy of Sciences of the Ukraine. Alterpres, Kyiv.

Chien, R. C., Lin, L. M., Chang, Y. H., Lin, Y. C., Wu, P. H., Asatiani, M. D., Wasser, S. G., Krakhmalnyi, M., Agbarya, A., Wasser, S. P., & Mau, J. L. (2016). Anti-inflammation properties of fruiting bodies and submerged cultured mycelia of culinary-medicinal higher Basidiomycetes mushrooms. International Journal of Medicinal Mushrooms, 18(11), 999–1009.

Classic Enzyme: Horseradish Peroxidase (2018). Journal of Chemical, Environmental and Biological Engineering, 2(2), 52–59.

Elisashvili, V., Kachlishvili, E., & Asatiani, M. D. (2018). Efficient production of lignin-modifying enzymes and phenolics removal in submerged fermentation of olive mill by-products by white-rot Basidiomycetes. International Biodeterioration and Biodegradation, 134, 39–47.

Fedotov, O. V. (2017). Condition of the prooxidant-antioxidant system of some strains of Basidiomycetes. Regulatory Mechanisms in Biosystems, 8(1), 77–83.

Fedotov, O. V., & Bisko, N. A. (2018). Effect of phenolic substances and hydrogen peroxide on antioxidant activity of some strains of Basidiomycetes. Innovative Biosystems and Bioengineering, 2(1), 4–10.

Fedotov, O. V., & Usikova, Z. L. (2020). The study of vegetative incompatibility strains of Basidiomycetes. Innovative scientific researches: European development trends and regional aspect. Baltija Publishing, Riga. Pp. 136–155.

Fedotov, O. V., & Velygodska, A. K. (2016). Milk-clotting and antioxidant activity of enzyme preparations of fungi strains of the order Polyporales s.l. The scientific heritage. Biological Sciences, 2(2), 71–76.

Fedotov, O. V., & Voloshko, T. E. (2014). Otrymannya ta analiz fermentnykh preparativ oksydoreduktaz deyakykh bazydiomitsetiv [Production and analysis of enzyme preparations of oxidoreductases of some Basidiomycetes]. Microbiology and Biotechnology, 3, 65–76 (in Ukrainian).

Gurung, N., Ray, S., Bose, S., & Rai, V. (2013). A broader view: Microbial enzymes and their relevance in industries, medicine and beyond. BioMed Research International, 2013, 1–18.

Hameed, A., Divine, M. S., Mazhar, K., Sm Faysal, B., & Sm Saker, B. (2018). Structure, function and applications of a classic enzyme: Horseradish peroxidase. Journal of Chemical, Environmental and Biological Engineering, 2(2), 52–59.

Hassan, W., Noreen, H., Rehman, S., Gul, S., Kamal, M. A., Kamdem, J. P., Zaman, B., & da Rocha, J. B. T. (2017). Oxidative stress and antioxidant potential of one hundred medicinal plants. Current Topics in Medicinal Chemistry, 17(12), 1336–1370.

Lushchak, V. I. (2016). Time-course and intensity-based classifications of oxidative stresses and their potential application in biomedical, comparative and environmental research. Redox Report, 21(6), 262–270.

Metri, Y., Warly, L., Metri, Y., Fachry, A. R., & Astuti, P. (2018). The enzymic degradation of lignin by white-rot fungi. Pakistan Journal of Nutrition, 17, 71–75.

Mhamdi, A., Queval, G., Chaouch, S., Vanderauwera, S., Van Breusegem, F., & Noctor, G. (2010). Catalase function in plants: A focus on Arabidopsis mutants as stress-mimic models. Journal of Experimental Botany, 61(15), 4197–4220.

Mittler, R. (2002). Oxidative stress, antioxidants and stress tolerance. Trends Plant Science, 7, 405–410.

Nandi, A., Yan, L. J., Jana, C. K., & Das, N. (2019). Role of catalase in oxidative stress- and age-associated degenerative diseases. Oxidative Medicine and Cellular Longevity, 2019, 9613090.

Singh, R., Kumar, M., Mittal, A., & Mehta, P. K. (2016). Microbial enzymes: Industrial progress in 21st century. Biotechnology, 6, 1–15.

Uneyama, H., & Takeuchi, K. (2012) New therapeutic strategy for amino acid medicine: Preface. Journal of Pharmacological Sciences, 118(2), 129–130.

Vitak, T. Y., Wasser, S. P., Nevo, E., & Sybirna, N. O. (2017). Enzymatic system of antioxidant protection of erythrocytes in diabetic rats treated with medicinal mushrooms Agaricus brasiliensis and Ganoderma lucidum (Agaricomycetes). International Journal of Medicinal Mushrooms, 19, 697–708.

Voloshko, T. E., & Fedotov, O. V. (2011). Skryninh shtamiv bazydiomitsetiv za aktyvnistyu antyoksydantnykh oksydoreduktaz [Screening of basidiomycetes strains on the antioxidant activity of oxidoreductases]. Microbiology and Biotechnology, 16, 69–81 (in Ukrainian).

Wasser, S. P. (2011). Current findings, future trends, and unsolved problems in studies of medicinal mushrooms. Applied Microbiology and Biotechnology, 89, 1323–1332.

Wasser, S. P. (2017). Medicinal mushrooms in human clinical studies. Part I. Anticancer, oncoimmunological, and immunomodulatory activities: A review. International Journal of Medicinal Mushrooms, 19(4), 279–317.

Yoshizawa, F. (2004). Regulation of protein synthesis by branched-chain amino acids in vivo. Biochemical and Biophysical Research Communications, 313, 417–422.

Yoshizawa, F. (2012). New therapeutic strategy for amino acid medicine: Notable functions of branched chain amino acids as biological regulators. Journal of Pharmacological Sciences, 118(2), 149–155.

How to Cite
Fedotov, O. V., & Usikova, Z. L. (2020). Modification of isolation methods and physico-biochemical properties of preparations of fungal oxidoreductases . Regulatory Mechanisms in Biosystems, 11(2), 310-314.