植物過氧化酶 (EC 1.11.17) 是屬於同功異構酶的家族。 這些同功異構酶的胺基酸序列有50% 以上的不同。 過氧化酶可作為IAA (植物生長素) 氧化酶。 以純化後的山葵 (Wasabia japonica) 過氧化酶進行動力分析，以IAA為受質的情況下，其對受質的親和力為 17.1 μM。針對以syringalazine (木質素單體的類似物) 為受質的情況下，親和力為80.7 μM 。以二維電泳純化後的山葵蛋白質，用LC/MS/MS (液相層析質譜儀) 做蛋白質定序，其胺基酸序列跟蛋白質資料庫比對，比對結果與阿拉伯介 (Arabidopsis thaliana)
果有10% 相似。

The plant peroxidases (EC1.11.17) exit as a large family of isozymes. These isozymes have more than 50% amino acid sequence differences. The function of Wasaba japonica peroxides plays the role as IAA oxidases. The kinetics result shows Wasabia japonica peroxidases displayed affinity (Km = 17.1 μM) for IAA. The kinetics results in Wasabia japonica peroxidases display affinity (Km = 80.6 μM) for syringaldazine. LC/MS/MS technique described the data that has proven to be a method for identification and characterization of proteins. The soluble proteins extracted form Wasabia japonica was purified by gel filtration chromatography and two-dimensional gel electrophoresis (2-DE). LC-MS/MS analyses of 2-DE gel spots and identify proteins structure based on the protein fragmentation characteristics. The Mascot Search Results showed that Wasabia japonica peroxidase has a significant similarity (10%) with Arabidopsis thaliana peroxidase.

Chinese abstract ----------------------------------- I

English Abstract ---------------------------------- II

Table of contents ------------------------------- III

List of figures ----------------------------------- IV

Introduction ------------------------------------- 1

Materials and Methods ------------------------ 5

Results ------------------------------------------- 10

Discussion --------------------------------------- 12

References --------------------------------------- 15

Figures ------------------------------------------- 19

Reference
Berglund, G.I., Carlsson, G.H., Smith, A.T., Szo¨ ke, H., Henriksen,
A., Hajdu, J., (2002) The catalytic pathway of horseradish peroxidase at high resolution. Nature 417, 463–468.
Christensen JH, Bauw G, Welinder KG, Van Montagu M, and Borerjan W (1998) Purification and Characterization of peroxidase correlated with lignification in popar xylem. Plant Physiol 118: 125-135
Dawson JH (1992) Probing structure-function relations in hem-containing oxygenases and peroxidase. Science 240; 433-439
Depree J. A., Howard T. M., and Savage G. P. (1998) Flavour and pharmaceutical properties of the volatile sulphur compounds of Wasabi (Wasabia japonica). Food Research International, 31:329-337.
Depree, J. A., Howard, T. M., and Savage, G. P. recently (1999) reported in Food Research International (31:329-337). Flavour and pharmaceutical properties of the volatile sulphur compounds of Wasabi (Wasabia japonica).
Duroux, L., Welinder, K.G., (2003) The peroxidase gene family in plants: a phylogenetic overview. J. Mol. Evol. 57, 397–407.
F’Oarrel, P.Z. (1975) High-resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007-4021.
Fuke Y., Ohishi Y., Iwashita K., Ono H., and Shinohara K. (1994) Growth suppression of MKN-28 human stomach cancer cells by Wasabi (Eutrema wasabi Maxim.). Journal of the Japanese Society for Food Science & Technology, 41:709-711.
Fuke Y., Haga Y., Ono H., Nomura, T., and Ryoyama K. (1997) Anti-carcinogenic activity of 6-methylsulfinylhexyl isothiocyanate-, an active anti-proliferative principal of Wasabi (Eutrema wasabi Maxim.). Cytotechnology, 25:197-203.
Gajhede, M., Schuller, D.J., Henriksen, A., Smith, A.T., Poulos, T.L.
(1997) Crystal structure of horseradish peroxidase C at 2.15
angstrom resolution. Nature Struct. Biol. 4, 1032–1038.
Gazaryan IG, Lagrimini LM (1996) Purification and unusual kinetic properties of a tobacco anionic peroxidase. Phytochemistry 41, 1029-34.
Gazaryan, I.G. and Lagrimini, L.M. (1996) Phytochemistry 42: 1271–1278.
Gazaryan I.G, lagrimini L.M, Ashby G.A., Thorneley R.N.F, Mechanism of iodole-3-acetic acid oxidation by plant peroxidase: anerobic stopped-flow spectrophotometric studies
Gazaryan I.G, Chubar T.A., Mareeva E.A., Largrimini L.M., Van Huystee R.B., Thorneley R.N.F., Aerobic oxidation of indole-3-acetic acid catalysed by anionic and cationic peanut peroxidase, Phytochemistry 51 (1999) 175-186
Hinman, R.L. and Lang, J. (1965) Biochemistry 4: 144–158.
on horseradish and tobacco peroxidases, Biochem.J.313 (1996) 841-847
Hiraga Susumu, Sasaki Katsutomo, Sasaki Hiroyuki, Ohashi YuKo, and Matsui Hirokazu (2001) A Large Family Class III Plant Peroxidases, Plant Cell Physiol. 42(5): 462-468
Lagrimini LM, Rothstein S (1987) Tissue Specificity of Tobacco Peroxidase Isozymes and Their Induction by Wounding and Tobacco Mosaic Virus Infection. Plant Physiol. 84, 438-442.
Lagrimini LM, Joly RJ, Dunlap JR, Liu TT (1997b) The consequence of peroxidase overexpression in transgenic plants on root growth and development. Plant Mol. Biol. 33, 887-95.
Lagrimini LM, Bradford S, Rothstein S. (1990) Peroxidase-Induced Wilting in Transgenic Tobacco Plants. Plant Cell 2, 7-18.
Mansouri, I.E., Mercado, J.A., Santiago-D