Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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Purification and Characterization of Polyphenol Oxidase in the Flesh of the Fuji Apple

  • Published : 2006.04.30
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Abstract

Polyphenol oxidase (PPO) was isolated from the flesh of Fuji apples by DEAE-Cellulose, ammonium sulfate precipitation, phenyl-Sepharose CL-4B, and Sephdex G-100 chromatography. The molecular mass of the purified PPO was estimated to be 40 kDa by SDS polyacrylamide gel electrophoresis. With regard to substrate specificity, maximum activity was achieved with chlorogenic acid as substrate, followed by catechin and catechol whereas, there was no detectable activity with hydroquinic acid, resorcinol, or tyrosine as substrate. The optimum pH and temperature with catechol as substrate were 6.5 and $35^{\circ}C$, respectively. The enzyme was most stable at pH 6.0 and unstable at acidic pH. The enzyme was stable when it was heated to $45^{\circ}C$ but heating at $50^{\circ}C$ for more than 30 min caused 50% loss of activity. Reduced $ZnSO_4$, L-cystein, epigallocatechin-3-o-gallate (EGCG), and gallocatechin gallate (GCG) also inhibited activity.

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References

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