Purification of d'Anjou Pear (Pyrus communis L.) Polyphenol Oxidase
作者： Kimberly W. Wissemann , Morris W. Montgomery
DOI: 10.1104/PP.78.2.256
关键词: Polyacrylamide 、 Hydroxylapatite 、 Pyrus communis 、 Polyphenol oxidase 、 PEAR 、 Chromatography 、 Enzyme assay 、 Gel electrophoresis 、 Chemistry 、 Pigment 、 Plant science 、 Genetics 、 Physiology
摘要: Polyphenol oxidase (PPO) was extensively purified to homogeneity from d9Anjou pear ( Pyrus communis L.) by extraction in the presence of phenolic binder AG 2-X8 andTriton X-100. Chlorophyll pigment removed chromatography resulting a clear, colorless enzyme extract. Purification PPO achieved after on Phenyl Sepharose CL-4B, DEAE-cellulose, and hydroxylapatite columns. Only columns were run at room temperature rather than 4°C sharp peaks good resolution obtained. Reproducibility entire scheme excellent with hydrophobic resin as key successful purification. Three separate fractions homogeneous when stained for protein silver stain polyacrylamide slab gel electrophoresis corresponded relative mobilities 0.41, 0.43, 0.73. The effect dimethylsulfoxide activity investigated found increase significantly over control.
plantphysiol.org
oup.com参考文章(19)
Mark A. Einerson, Effect of cyclopropenoid fatty acids on membrane components of liver of rainbow trout (Salmo gairdneri) ,(1982)
Denise Myrtle Smith, Methods of extraction and characterization of polyphenol oxidase from d'Anjou pears (Pyrus Communis L.) ,(1979)
Stellan Hjertén, Jan Rosengren, Sven P»hlman, Hydrophobic interaction chromatography Journal of Chromatography A. ,vol. 101, pp. 281- 288 ,(1974) , 10.1016/S0021-9673(00)82845-9
William H. Flurkey, Joseph J. Jen, Purification and characterization of polyphenol oxidase in redhaven peaches Biochemie und Physiologie der Pflanzen. ,vol. 175, pp. 637- 642 ,(1980) , 10.1016/S0015-3796(80)80017-5
Miriam Kidron, Eitan Harel, Alfred M. Mayer, Copper content and amino acid composition of catechol oxidase from clairette grapes Phytochemistry. ,vol. 16, pp. 1050- 1051 ,(1977) , 10.1016/S0031-9422(00)86724-2
WILLIAM H. FLURKEY, JOSEPH J. JEN, PEROXIDASE AND POLYPHENOL OXIDASE ACTIVITIES IN DEVELOPING PEACHES Journal of Food Science. ,vol. 43, pp. 1826- 1828 ,(1978) , 10.1111/J.1365-2621.1978.TB07424.X
Wayne Wray, Teni Boulikas, Virginia P. Wray, Ronald Hancock, Silver staining of proteins in polyacrylamide gels. Analytical Biochemistry. ,vol. 118, pp. 197- 203 ,(1981) , 10.1016/0003-2697(81)90179-2
R.J. Yon, Recent developments in protein chromatography involving hydrophobic interactions International Journal of Biochemistry. ,vol. 9, pp. 373- 379 ,(1978) , 10.1016/0020-711X(78)90048-4
P.W. Holloway, A simple procedure for removal of triton X-100 from protein samples Analytical Biochemistry. ,vol. 53, pp. 304- 308 ,(1973) , 10.1016/0003-2697(73)90436-3
Kimberly W. Wissemann, C.Y. Lee, Purification of grape polyphenoloxidase with hydrophobic chromatography Journal of Chromatography A. ,vol. 192, pp. 232- 235 ,(1980) , 10.1016/S0021-9673(00)81860-9