Abstract
This study aimed to extract and purify polyphenol oxidase enzyme (PPO) from mushrooms and use it in pastry industry. The enzyme was extracted from edible mushroom using different buffers and purified by several steps. Some Characterizations of the isolated PPO were studied. The activity of PPO was evaluated using spectrophotometric method and catechol as a substrate. Characterization studies indicated that the enzyme showed the highest specific activity 2087 U/mg protein at pH 6.0 using 0.2M phosphate buffer. The ratio of 1:2 (w:v) from mushroom: buffer was the best for obtaining the highest specific activity (2490 U/mg protein). The optimum time of extraction of PPO was 10 mins to obtain high specific activity (2625 U/mg protein). The precipitation of ammonium sulfate and ion-exchange chromatography techniques were performed to purify PPO. The precipitation with saturated solution of ammonium sulfate (70%) was used, the Specific activity was 4332.7 U/mg protein with purification fold of 1.7 and enzymatic yield of 65.5%. Ion exchange chromatography using DEAE cellulose column estimated that the purification times of the enzymatic extract were 6.01, with an enzymatic yield 22.3%. Storage study of the PPO in edible mushroom at 4oC for 3 months indicated that the PPO loss the stability gradually over time, 33% from its original activity retained after 32 days. It can be concluded that edible mushroom is a good source of PPO and the enzyme still active when the mushroom was stored for one month.
Published Version
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