The effect of horseradish peroxidase (HRP) and its dextran derivative on environment pollutant dyes

Horseradish Peroxidase (HRP; E.C 1.11.1.7) notably used in diagnostic industry and having a great commercial importance, is extracted from the roots of Armporacia rusticana. HRP enzyme is a heme containing peroxidase which has been studied for a century and has so many isoenzymes. Enzymatic activity of HRP take its source from the reduction and oxidation of iron atom present in heme group. As a result of having a heme group, HRP has an absorbance at 403 nm . Peroxidases are oxidoreductases which use hidrogen peroxidase or organic hydrogen peroxidases as oxidants. Most of them are glycoprotein structured enzymes having N-linked oligosaccaharides in their compositions. They have three different forms known as Compound I, II and III with H2O2.Fenolic compaunds are major pollutants in medicine, food and local matrixe.Hence, it is important to detect fenolic compounds sensitively. Horseradish peroxidase (HRP) can be catalyse oxidation of heteroaromatic compounds as phenols, biphenols, alinine. Because of HRP shows wide range of pH and temparature, it is seem favor to use processsing to waste water. In recent 20 years in order to optimize the removal of phenol and analogous from waste water, many studies have been performed. It is known that HRP catalyse free radical existion before polymerization of phenol, chlorophenol and other substitue phenols.H2O2 + 2 AH2 HRP 2AH + 2H2OVarious methods were tested such as chemical oxidation, reverse osmos, adsorpsition to remove these toxic dyes. Recently, investigators who work in this area focus on removing these toxic substances by enzyme catalyses. Especially, it is emphasized on plant peroxidases.In this study, Horseradish Peroxidase-Dextran covalent conjugate was synthesized. Thus, firstly HRP was purified by affinity chromatography using immobilized Con A. Enzyme-dextran conjugate with 1/10 molar ratio was synthesized using dextran aldehyde derivative (Dextran Mw 75.000 Da) and purified enzyme.Bleaching of Naphtol Blue Black and Coomassie Brillant Blue dyes were observed with UV-VIS absorbance spectrophotometer for 60 minutes and results are compared. The reactions by using purified enzyme and conjugate were performed at different pHs (pH: 3, 4, 5, 6, 7, 8) in a 25, 30, 35, 40, 45, 50, 60, 70, oC temperatured in UV-VIS quartz cuvette. Degredation of ?Naphtol Blue Black? and ?Coomassie Brillant Blue? were evaluated using LC-MS and FT-IR spectra.