Polyphenol oxidase from wheat bran is a serpin

• Autorzy: Yamasaki Y. , Konno H. , Noda K.
• Języki publikacji: EN

Abstrakty

EN

Polyphenol oxidase (PPO; EC 1.10.3.2) was isolated from wheat bran by a procedure that included ammonium sulfate fractionation, batch adsorption by DEAE-cellulofine, CM-cellulofine column chromatography, DEAE-cellulofine column chromatography, preparative isoelectric focusing, adsorption on the membrane of a Vivapure Q Maxi H spin column, and heat treatment. These procedures led to 150-fold purification with 4.2% recovery. The PPO was homogeneous by SDS/PAGE. The relative molecular weight of the PPO was estimated to be 37000 based on its mobility in SDS/PAGE. The isoelectric point of the PPO was 4.4. The Kmvalues of the PPO for caffeic acid, chlorogenic acid, pyrocatechol, 4-methyl catechol and l-DOPA as substrates were 0.077, 0.198, 1.176, 1.667 and 4.545 mM. The PPO was strongly inhibited by tropolone. The Kivalue for tropolone is 2.2 × 10–7M. The sequence of the 15 N-terminal amino-acid residues was determined to be ATDVRLSIAHQTRFA, which was identical to those of serpin from Triticum aestivum and protein Z from Hordeum vulgare. The PPO strongly inhibited the activity of trypsin, which is an enzyme of serine proteases; 50% inhibition was observed with 1.5 × 10–7M PPO. The Kivalue for PPO is 2.3 × 10–8M. The wheat bran PPO should be a very important protein for protecting wheat against disease, virus, insect and herbivore damages by both the activities of PPO and protease inhibitor.

Słowa kluczowe

PL

wheat bran; polyphenol oxidase; serpin; purification; general property; chelating reagent; substrate specificity

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2008
• Tom: 55
• Numer: 2
• Opis fizyczny: p.325-328,fig.,ref.

Twórcy

autor

Yamasaki Y.

• Okayama University, Kurashiki-shi, Okayama 710-0046, Japan

autor

Konno H.

autor

Noda K.

Bibliografia

• Awdeh ZL, Williamson AR, Askonas BA (1968) Isoelectric focusing in polyacrylamide gel and its application to immunoglobulins. Nature 219:66-67.
• Brandt A, Svendsen I, Hejgaard J (1990) A plant serpin gene. Structure, organization and expression of the gene encoding barley protein Z. Eur J Biochem 194:499-505.
• Chen JS, Wei CI, Marshall M (1991) Inhibition mechanism of kojic acid on polyphenol oxidase. J Agric Food Chem 39:1897-1901.
• Fawcett JS (1968) Isoelectric fractionation of proteins on polyacrylamide gels. FEBS Lett 1:81-82.
• Felton GW, Donato K, Del Vecchio RJ, Duffey SS (1989) Activation of plant polyphenol oxidases by insect feeding damage reduces nutritive quality of foliage for noctuid herbivores. J Chem Ecol 15:2667-2694.
• Felton GW, Donato K, Del Vecchio RJ, Duffey SS (1992) Impact of oxidized plant phenolics on the nutritional quality of dietary protein to a noctuid herbivore. J Insect Physiol 38:277-285.
• Hejgaard J, Rasmussen SK, Brandt A, Svendsen I (1985) Sequence homology between barley endosperm protein Z and protease inhibitors of the alpha-1-antitrypsin family. FEBS Lett 180:89-94.
• Hind G, Marshak D, Coughalan S (1995) Spinach thylakoid polyphenol oxidase: cloning, characterization, and relation to a putative protein kinase. Biochemistry 34:8157-8164.
• Kahn V, Andrawis A (1985) Inhibition of mushroom tyrosinase by tropolone. Phytochemistry 24:905-908.
• Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature 227:680-685.
• Li L, Steffens J C (2002) Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance. Planta 215:239-247.
• Mazzafera P, Robinson SP (2000) Characterization of polyphenol oxidase in coffee. Phytochemistry 55:285-296.
• Melo GA, Shimizu MM, Mazzafera P (2006) Polyphenoloxidase activity in coffee leaves and its role in resistance against the coffee leaf miner and coffee leaf rust. Phytochemistry 67:277-285.
• Moore BM, Flurkey WH (1990) Sodium dodecyl sulfate activation of a plant polyphenoloxidase: effect of sodium dodecyl sulfate on enzymatic and physical characteristics of purified broad bean polyphenol oxidase. J Biol Chem 265:4982-4988.
• Nielsen G, Johansen H, Jensen J, Hejgaard J (1983) Localization on barley chromosome 4 of genes coding for beta-amylase (Bmyl) and protein Z (Pazl). Barley Genet Newsl 13:55-57.
• Rasmussen SK, Dahl SW, Norgard A, Hejgaard J (1996) A recombinant wheat serpin with inhibitory activity. Plant Mol Biol 30:673-677.
• Ray H, Hammerschmidt R (1998) Responses of potato tuber to infection by Fusarium sambucinum. Physiol Mol Plant Pathol 53:81-92.
• Ostergaard H, Rasmussen SK, Roberts TH, Hejgaard J (2000) Inhibitory serpins from wheat grain with reactive centers resembling glutamine-rich repeats of prolamin storage proteins - Cloning and characterization of five major molecular forms. J Biol Chem 275:33272-33279.
• Shi C, Dai Y, Xu X, Xie Y, Liu Q (2002) The purification of polyphenol oxidase from tobacco. Protein Expr Purif 24:51-55.
• Steffens JC, Harel E, Hunt MD (1994) Polyphenol oxidase. In: Recent Advances in Phytochemistry, Genetic Engineering of Plant Secondary Metabolism. Ellis BE, Kuroki GW, Stafford HA, eds, vol 28, pp 275-312. Plenum Press, New York.
• Swain T, Mapson IW, Robb DA (1966) Activation of Vicia faba (L) tyrosinase as effected by denaturing agents. Phytochemistry 5:469-482.
• Trebst A, Depka B (1995) Polyphenol oxidase and photosynthesis research. Photosynthesis Res 46:41-44.
• Wang J, Constabel CP (2003) Biochemical characterization of two differentially expressed polyphenol oxidases from hybrid poplar. Phytochemistry 64:115-121.
• Warburg O, Christian W (1942) Isolation and crystallization of enolase. Biochem Z 310:384-421.
• Wititsuwannakul D, Chareonthiphakorn N, Pace M, Wititsuwannakul R (2002) Polyphenol oxidases from latex of Hevea brasiliensis: purification and characterization. Phytochemistry 61:115-121.