Haloperoxidase activity of manganese peroxidase from Phanerochaete chrysosporium.

Manganese peroxidase (MnP) from Phanerochaete chrysosporium exhibits haloperoxidase activity at low pH. In the presence of hydrogen peroxide, MnP oxidizes bromide and iodide as measured by the formation of tribromide and triiodide complexes and the halogenation of various organic substrates. The optimum pHs for bromide and iodide oxidation are 2.5 and 3.0, respectively. Transient-state kinetic studies show that the reaction between MnP compound I and bromide or iodide occurs via a single two-electron step process, obeying second-order kinetics. The second-order rate constants for MnP compound I reduction by bromide and iodide are (4.1 +/- 0.2) x 10(3) and (1.1 +/- 0.1) x 10(5) m-1 s-1, respectively, at pH 3.0. MnP brominates a variety of aromatic substrates, including veratryl (3,4-di-methoxybenzyl) alcohol (I) to produce of 2-bromo-4,5-dimethoxybenzyl alcohol (II). MnP also hydrobrominates cinnamic acid (VI) to produce 2-bromo-3-hydroxyphenylpropionic acid (VII). With 3,4-dimethoxycinnamic acid (III) as the substrate, two bromination products are identified: trans-2-bromo-1-(3, 4-dimethoxyphenyl) ethylene (IV) and 2-bromo-3-(3, 4-dimethoxyphenyl)-3-hydroxypropionic acid (V). MnP also brominates 1,3-dicarbonyl compounds such as monochlorodimedone and malonic acid. Incubation of MnP with bromide and H2O2 in the absence of organic substrates results in enzyme inactivation. MnP binds halides to produce characteristic optical difference spectra. From these spectra, apparent dissociation constants at pH 3.0 are determined to be 0.13, 20, and 45 mm for fluoride, chloride, and bromide, respectively.