Browse > Article

Purification and Characterization of Polyphenol Oxidase from Lotus Root (Nelumbo nucifera G.)  

Moon, Sang-Mi (Department of Food Biotechnology, Mokpo National University)
Kim, Hyun-Jin (Department of Food Science and Human Nutrition, Clemson University)
Ham, Kyung-Sik (Department of Food Biotechnology, Mokpo National University)
Publication Information
Korean Journal of Food Science and Technology / v.35, no.5, 2003 , pp. 791-796 More about this Journal
Abstract
Polyphenol oxidase isoforms were purified from the lotus roots using 50% acetone precipitation, conventional chromatographies of Q-Sepharose and hydrophobic interaction, and high performance liquid chromatographies of Mono-Q and Superdex 75 gel-filtration. Molecular mass of a purified PPO isoform (LPIII-2) was determined to be 56 kDa using gel-filtration chromatography. The active form of LPIII-2 appeared to bea heterodimer, as purified LPIII-2 on SDS-PAGE gel showed two bands that were determined to be 28 kDa and 26 kDa. To further characterize PPO, partially purified PPO isoforms (LP-II, LP-III) were obtained from Q-Sepharose anion-exchange chromatography. In substrate specificity, the partially purified PPO isoform LP-II showed a high affinity to catechol, while LP-III showed a high affinity to pyrogallol. The optimum pH of LP-II and LP-III was pH 7.0. Interestingly, the partially purified PPO isoforms showed high activities at low temperatures $(0{\sim}5^{\circ}C)$, and as temperatures rose, the activities decreased. Both PPO isoforms were stable at $40^{\circ}C$ and were inactivated by incubation at $60^{\circ}C$ for 40 min.
Keywords
lotus root; polyphenol oxidase; purification; characterization;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Park, S.-Y., Hwang, T.-Y., Kim, J.-H. and Moon, K.-D. Quality changes of minimally processed lotus root (Nelumbo nucifera) with browning inhibitors. Korean J. Postharvest Sci. Technol. 8: 164-168 (2001)
2 Kim, D.M. Minimal processing technology of fruits and vegetables. Food Sci. Ind. 30: 95-102 (1997)
3 Jang, J.-S. and Hong, G.-H. The effects of storage temperature and pH on color change in garlic puree. Korean J. Postharvest Sci. Technol. 5: 211-216 (1998)
4 Choi, S.-T., Chang, K.-S., Lim, B.-S., Lee, C.-S. and Kim, Y.-B. Effect of storage and marketing condition on biochemical property changes of garlic (Allium sativum L.). Korean J. Postharvest Sci. Technol. 5: 111-117 (1998)
5 Espm, J.C., Morales, M., Gracin-Ruiz, P.A., Tedela, J. and Garcia-Canovas, F. Improvement of a continuous spectrophotometric method for determining the monophenolase and diphenolase activities of mushroom polyphenol oxidase. J. Agric. Food Chem. 45: 1084-1090 (1997)   DOI   ScienceOn
6 Rathjen, A.H. and Robinson, S.P. Aberrant processing of polyphenol oxidase in a variegated grapevine mutant. Plant PhysioL 99: 1619-1625 (1992)   DOI   ScienceOn
7 Sanchez-Ferrer, A., Laveda, F. and Garcia-Carmona, F. Substratedependent activation of latent leaf polyphenol oxidase by anionic surfactants. J. Agric. Food Chem. 41: 1583-1586 (1993)   DOI   ScienceOn
8 Moore, B.M. and Flurkey, W.H. Sodium dodecyl sulfate activation of a plant polyphenol oxidase: Effect of sodium dodecyl sulfate on enzymatic and physical characteristics of purified broad bean polyphenol oxidase. J. BioI. Chem. 265: 4982-4988 (1990)
9 Donald, V. and Judith, G.V. Biochemistry, pp. 345-370. John Wiley & Sons, Inc., New York, USA (1995)
10 Sodfrhall, J. Properties of carrot polyphenol oxidase. Phytochemistry 39: 33-38 (1995)   DOI   ScienceOn
11 Espin, J.C., van Leeuwen, J. and Wichers, H.J. Kinetic study of the activation process of a latent mushroom (Agaricus bisporus) tyrosinase by serine proteases. J. Agric. Food Chem. 47: 3509-3517 (1999)   DOI   ScienceOn
12 Kubo, E., Kinst-Hori, I., Kubo, Y., Yamagiwa, Y., Kamikawa, T. and Haraguchi, H. Molecular design of antibrowning agents. J. Agric. Food Chem. 48: 1393-1399 (2000)   DOI   ScienceOn
13 Kim, D.M. Extension of freshness of minimally processed fruits and vegetable. Korean J. Hort. Sci. Technol. 17: 790-795 (1999)
14 Valero, E. and Garcia-Carmona, F. pH-dependent effect of sodium chloride on latent grape polyphenol oxidase. J. Agric. Food Chem. 46: 2447-2451 (1998)   DOI   ScienceOn
15 Ham, K.-S., Kauffmann S., Albersheim, P. and Darvill, A.G. Host-pathogen interactions XXXIX. A soybean pathogenesisrelated protein with $\beta$-1,3-glucanase activity releases phytoalexin elicitor-active heat-stable fragments from fungal walls. Mol. Plant-Microbe Interactions 4: 545-552 (1991)   DOI
16 Kim, D.-Y., Rhee, C.-O. and Kim, Y.-B. Characteristics of polyphenol oxidase from garlic (Allium sativum L.). J. Korean Agric. Chem. Soc. 24: 167-173 (1981)
17 Kim, Y.-S., Jeon, S.-S. and Jung, S.-T. Effect of lotus root powder on the baking quality of wheat bread. Korean J. Soc. Food Cookery Sci. 18: 413-425 (2002)
18 Van Geledr, C.W.G., Flurkey, W.H. and Wichers, H.J. Sequence and structural features of plant and fungal tyrosinases. Phytochemistry 45: 1309-1323 (1997)   DOI   ScienceOn
19 Kim, Y.-S., Chun, S.-S., Jung, S.-T. and Kim, R.-Y. Effect of lotus root powder on the quality of dough. Korean J. Soc. Food Cookery Sci. 18: 573-578 (2002)
20 Seo, J.H., Hwang, Y.S., Chun, J.P. and Lee, J.-C. Change of phenolic compounds and occurrence of skin browning, and characterization of partially purified polyphenol oxidases in oriental pear fruits. J. Korean Soc. Hort. Sci. 42: 184-188 (2001)
21 Burton, K.S., Love, M.E. and Smith, J.F. Biochemical changes associated with mushroom quality in Agaricus spp.. Enzyme Mirobiol. Technol. 15: 736-741 (1993)   DOI   ScienceOn
22 Thipyapong, P. and Steffens, J.C. Tomato polyphenol oxidase. Plant Physiol 115: 409-418 (1997)   PUBMED
23 Pyo, H.-F., Son, D.-Y. and Lee, C. Purification and characterization of polyphenol oxidase from Flammulina velutips. Korean J. Food Sci. Technol. 34: 552-558 (2002)
24 Mayer, A.M. Polyphenol oxidase in plants-recent progress. Phytochemistry 26: 11-20 (1987)   DOI   ScienceOn
25 Espin, J.C. and Wichers, H.J. Kinetics of activation of latent mushroom (Agaricus bisporus) tyrosinase by benzyl alcohol. J. Agric. Food Chem. 47: 3503-3508 (1999)   DOI   ScienceOn
26 Friedman, M. Chemistry, biochemistry, and dietary role of potato polyphenols. J. Agric. Food Chem. 45: 1523-1540 (1997)   DOI   ScienceOn
27 Van Gelder, C.W.G., Flurkey, W.H. and Wichers, H.J. Sequence and structural features of plant and fungal tyrosinases. Phytochemistry 45: 1309-1323 (1997)   DOI   ScienceOn
28 Burton, S.G. and Duncan, J.R. Activation of mushroom polyphenol oxidase in organic medium by the detergent SDS. Biotechnol. Lett. 17: 627-630 (1995)   DOI   ScienceOn
29 Kahn, V. Polyphenol oxidase isoenzymes in avocado. Phytochemistry 15: 267-272 (1976)   DOI   ScienceOn
30 Park, W.P., Cho, S.H. and Lee, D.S. Screening of antibrowning agents for minimally porcessed vegetable. Korean J. Food Sci. Technol. 30: 278-282 (1998)
31 Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254 (1976)   DOI   PUBMED   ScienceOn