Purification and partial characterization of haloperoxidase from fresh water algae Cladophora glomerata

Many haloperoxidases have been purified from diverse organisms, including l ichen, fungi, bacteria, and marine algae. In this study a haloperoxidase wa s purified from the fresh water algae. Cladophora glomerata, by homogenizat ion and centrifugation, ammonium sulfate fractionation, ion-exchange and ge l filtration chromatography. Molecular weight was determined by SDS-PAGE an d by size exclusion HPLC and found to be approximately 43 kDa. The isoelect ric point was determined to be approximately 8.1 by isoelectric focusing. T he UV spectrum of the peroxidase showed a strong absorbance in the Soret ba nd indicating a heme protein, unlike vanadium-dependent haloperoxidases fro m marine algae. Fresh water algal haloperoxidase catalyzed the iodination o f tyrosine at a pH of 3.1. This haloperoxidase also catalyzes the oxidation of guaiacol and oxidation of iodide as well as catalyzing a peroxide-depen dent reaction in both the presence and absence of chloride and bromide ions . (C) 2000 Published by Elsevier Science Inc. All rights reserved.