Abstract:
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.
