Purification and some kinetic properties of catalase from parsley (Petroselinum hortense Hoffm., Apiaceae) leaves.
In this study, catalase (CAT: EC 1.11.1.6) was purified from parsley (Petroselinum hortense) leaves; analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps, including preparation of homogenate, ammonium sulfate fractionation, and fractionation by DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 9.5% and had a specific activity of 1126 U (mg proteins)(-1). The overall purification was about 5.83-fold. A temperature of 4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured at 240 nm. In order to control the purification of the enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acryl amide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for the enzyme. The molecular weight was found to be 183.29 kDa by Sephadex G-200 gel filtration chromatography. The stable pH, optimum pH, and ionic strength were determined for phosphate and Tris-HCl buffer systems. In addition, K(M) and V(max) values for H(2)O(2), at optimum pH and 25 degrees C, were determined by means of Lineweaver-Burk plots.
Oztürk
Lokman
L
Faculty of Science and Arts, Department of Biology, Gaziosmanpasa University, Tokat, Turkey. lokmanoz@gop.edu.tr
Bülbül
Metin
M
Elmastas
Mahfuz
M
Ciftçi
Mehmet
M
D000645:Ammonium Sulfate / Q000737:chemistry; D002374:Catalase / Q000302:isolation & purification* / Q000378:metabolism*; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D028528:Petroselinum / Q000201:enzymology*; D018515:Plant Leaves / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D002374:Catalase; D000645:Ammonium Sulfate