Abstract
Polyphenol oxidase from the latex of opium poppy was purified to the electrophoretic homogeneity by affinity chromatography using p-aminobenzoic acid as a ligand coupled to Sepharose CL-4B by divinyl sulphone activation method. The purified enzyme was used to prepare the polyclonal antibodies. The purified latex PPO exhibited high diphenolase activity in comparison with almost unmeassurable monophenolase activity. Both of these activities were sensitive to the activation with sodium dodecyl sulphate. Two isoforms (65 and 40 kDa) of latex PPO were separated by the gel filtration. There were no differences in substrate specifity (weak monophenolase and high diphenolase activity) and sensitivity to inhibitors between these isoforms, but they showed differences in electrophoretic mobility.
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References
Bilka, F., Bala zová, A., Bilková, A., Pšenák, M.: Purification of polyphenol oxidase from opium poppy latex.-Pharmazie 55: 155-156, 2000.
Bilková, A., Bezáková, L., Bilka, F., Bala ová, A., Pšenák, M.: [Amine oxidase in biosynthesis of opium alkaloids in seedlings of Papaver somniferum L.]-Chem. Listy 94: 739, 2000a. [In Slovak.]
Bilková, A., Bezáková, L., Bilka, F., Pšenák, M.: [Role of aminoxidase in alkaloid biosynthesis.]-Czech Slov. Pharm. 49: 171-176, 2000b. [In Slovak.]
Bradford, M.: A rapid and sensitive method for the quantication of microgram quantities of proteins utiliting the principle of protein-dye binding.-Anal. Biochem. 72: 248-254, 1976.
Chou, W.M., Kutchan, T.M.: Enzymatic oxidations in the biosynthesis of complex alkaloids.-Plant J. 15: 289-300, 1998.
Escribano, J., Cabanes, J., Chazarra, S., García-Carmona, F.: Characterization of monophenolase activity of table beet polyphenol oxidase. Determination of kinetic parameters on the tyramine-dopamine pair.-J. agr. Food Chem. 45: 4209-4214, 1997.
Facchini, P.J., Bird, D.A.: Developmental regulation of benzylisoquinoline alkaloid biosynthesis in opium poppy plants and tissue cultures.-In Vitro cell. dev. Biol.-Plant 34: 69-79, 1998.
Facchini, P.J., De Luca, V.: Differential and tissue-specific expression of a gene family for tyrosine/dopa decarboxylase in opium poppy.-J. biol. Chem. 269: 26684-26690, 1994.
Fujita, S., Saari, N., Maegawa, M., Tetsuka, T., Hayashi, N., Tono, T.: Purification and properties of polyphenol oxidase from cabbage (Brassica oleracea L.).-J. agr. Food Chem. 43: 1138-1142, 1995.
Jiménez, M., García-Carmona, F.: The effect of sodium dodecyl sulphate on polyphenol oxidase.-Phytochemistry 42: 1503-1509, 1996.
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4.-Nature 227: 680-685, 1970.
Leoni, O., Palmieri, S.: Polyphenol oxidase from Artichoke (Cynara scolymus L.).-Food Chem. 38: 27-39, 1990.
Moore, B.M., Flurkey, W.H.: Sodium dodecylsulfate activation of a plant polyphenoloxidase.-J. biol. Chem. 265: 4982-4988, 1990.
Nozue, M., Souri, M., Arakawa, D., Kojima, M.: Purification and characterization of two isoforms of chlorogenic acid oxidase from sweet potato cells in suspension culture.-J. Plant Physiol. 153: 552-557, 1998.
Park, S.U., Johnson, A.G., Penzes-Yost, C., Facchini, P.: Analysis of promoters from tyrosine/dihydroxyphenylalanine decarboxylase and berberine bridge enzyme genes involved in benzylisoquinoline alkaloid biosynthesis in opium poppy.-Plant mol. Biol. 40: 121-131, 1999.
Roberts, M.F.: Polyphenolases in the 1 000 g fractions of Papaver somniferum latex.-Phytochemistry 10: 3021-3027, 1971.
Rodriquez, M.O., Flurkey, H.W.: A biochemistry project to study mushroom tyrosinase.-J. chem. Educ. 69: 767-769, 1992.
Rueffer, M., Zenk, M.H.: Distant precursors of benzylisoquinoline alkaloids and their enzymatic formation.-Z. Naturforsch. 42c: 319-332, 1987.
Stadler, R., Kutchan, T.M., Zenk, M.H.: (S)-norcoclaurine is the central intermediate in benzylisoquinoline alkaloid biosynthesis.-Phytochemistry 28: 1083-1086, 1989.
Van Gelder, C.W.G., Flurkey, W.H., Wichers, H.J.: Sequence and structural features of plant and fungal tyrosinases.-Phytochemistry 45: 1309-1323, 1997.
Zichová, M., Švajdlenka, E., Pšenák, M.: The content and distribution of precursors and alkaloids in developing poppy seedlings.-Chem. Listy 9: 687-688, 1996a.
Zichová, M., Švajdlenka, E., Pšenák, M.: Content of precursors and alkaloids in opium poppy latex during capsule maturation.-Chem. Listy 9: 701-702, 1996b.
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Bilka, F., Balažová, A., Bilková, A. et al. Characterization of Polyphenol Oxidase from the Latex of Opium Poppy. Biologia Plantarum 47, 111–115 (2003). https://doi.org/10.1023/A:1027345318226
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DOI: https://doi.org/10.1023/A:1027345318226