Purification and Properties of Polyphenol Oxidase of Dried Volvariella bombycina
Sarsenova A. Demir D. Çağlayan K. Abiyev S. Darbayeva T. Eken C.
January 2023MDPI
Biology
2023#12Issue 1
Polyphenol oxidase (PPO) was purified and characterized from a dried wild edible and medicinal mushroom (V. bombycina). Using Sepharose 4B-L-tyrosine-p-aminobenzoic acid affinity chromatography, PPO was purified from the dried V. bombycina. The purification was completed with a 33.85-fold purification. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the purified enzyme migrated as a single band. The molecular weight of the purified enzyme was estimated by SDS-PAGE to be about 25 kDa. Catechol, 4-methyl catechol, and pyrogallol were used as substrates to determine the enzyme activity and its kinetic parameters (Km and Vmax). At the optimum pH and temperature, dried V. bombycina PPO’s Km and Vmax values for catechol, 4-methyl catechol, and pyrogallol were found to be 1.67 mM–833.33 U/mL, 3.17 mM–158.73 U/mL, and 2.67 mM–3333.33 U/mL, respectively. Also investigated were the effects of pH and temperature on the enzymatic properties of PPO in dried V. bombycina. The optimum pH and temperature values for dried V. bombycina PPO obtained by using catechol, 4-methyl catechol, and pyrogallol as substrates were 6.5, 15 °C; 9.0, 20 °C; and 8.0, 15°C, respectively. This is the first study on the purification and characterization of PPO from dried V. bombycina.
affinity chromatography , characterization , polyphenol oxidase , purification , Volvariella bombycina
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Department of General Biology and Genomics, Faculty of Natural Sciences, L.N. Gumilyov Eurasian National University, Satpayev str. 2, Astana, 010000, Kazakhstan
Department of Biology and Ecology, Faculty of Natural Geography, Makhambet Utemisov West Kazakhstan University, N. Nazarbayev ave.162, Uralsk, 090000, Kazakhstan
Department of Agricultural Biotechnology, Faculty of Agriculture, Isparta University of Applied Sciences, Isparta, 32260, Turkey
Department of Agricultural Biotechnology, Faculty of Agriculture, Aydın Adnan Menderes University, Aydın, 09070, Turkey
Department of General Biology and Genomics
Department of Biology and Ecology
Department of Agricultural Biotechnology
Department of Agricultural Biotechnology
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