Purification and characterisation of vanadium haloperoxidases from the brown alga Pelvetia canaliculata

Two enzymes characterised as iodoperoxidases (PcI and PcII), with vanadium-dependent activity, have been purified from the brown alga Pelvetia canaliculata (L.) Decne et Thur. (Fucaceae, Phaeophyceae), collected in the Northern Portuguese coast, at Viana do Castelo. The relative molecular masses were 166 kDa for PcI and 416 kDa for PcII, as determined by gel filtration. SDS-PAGE shows that PcI has just one band corresponding to a subunit of 66 kDa, while PcII shows four bands (66, 72, 157 and 280 kDa). The following kinetic parameters have been determined from a steady-state analysis of the oxidation of iodide by H²O²: PcI, pH(opt) = 6.0, K(M)(I^-) = 2.1 mM, K(M)(H₂O₂) = 110 μM, K(i)(I^-) = 127 mM; and PcII, pH(opt) = 6:5, K(M)(I^- ) = 2.4 mM, K(M)(H₂O₂) = 20 μM and K(i)(I^-) = 69 mM. These iodoperoxidases are thermostable, as also observed for vanadium bromo- and chloroperoxidases. (C) 2000 Elsevier Science Ltd.