Journal
JOURNAL OF GINSENG RESEARCH
Volume 37, Issue 1, Pages 117-123Publisher
KOREAN SOC GINSENG
DOI: 10.5142/jgr.2013.37.117
Keywords
Panax ginseng; Polyphenol oxidase; Purification; Catechol; Carboxymethyl-Sepharose
Funding
- Chung-Ang University Research Scholarship Grant
- 'Industry and Academy Collaboration for Gyeonggi-Ginseng' Program
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Polyphenol oxidase (PPO) was purified from fresh ginseng roots using acetone precipitation, carboxymethyl (CM)-Sepharose chromatography, and phenyl-Sepharose chromatography. Two isoenzymes (PPO 1 and PPO 2) were separated using an ion-exchange column with CM-Sepharose. PPO 1 was purified up to 13.2-fold with a 22.6% yield. PPO 2 bound to CM-Sepharose, eluted with NaCl, and was purified up to 22.5-fold with a 17.4% yield. PPO 2 was further chromatographed on phenyl-Sepharose. The molecular weight of the purified PPO 2 from fresh ginseng was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was about 40 kDa. The optimum temperature and pH were 20 degrees C and 7.0, respectively, using catechol as a substrate. Pyrogallol showed the highest substrate specificity. The effect of a PPO inhibitor showed that its activity increased slightly in the presence of a low concentration of citric acid. High concentrations of acidic compounds and sulfite agents significantly inhibited purified ginseng PPO 2.
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