title: Anionic tobacco peroxidase is active at extremely low pH: veratryl alcohol oxidation with a pH optimum of 1.8.
abstract: Tobacco peroxidase (36 kDa, pI 3.5) exhibits unique catalytic and spectral properties that are modulated by pH, calcium and magnesium ions. It catalyses the oxidation of veratryl alcohol by hydrogen peroxide over a wide pH range (1.5-5.0) in the presence of these metal ions with a pH optimum of 1.8. This is the only example of a holoperoxidase described so far that is active and comparatively stable at such a low pH. The enhancement of tobacco peroxidase activity by magnesium ions is to our knowledge the first example of a magnesium-induced peroxidase activation. UV/visible spectra of tobacco peroxidase showed that the Soret band shifted and its absorption coefficient increased upon the addition of calcium or magnesium ions and on lowering the pH. The tobacco peroxidase spectrum at pH 1.85, in the presence of calcium chloride (> 50 mM), is similar to that of lignin peroxidase at pH 6.0, with the Soret band shifting from 403 to 409 nm and the molar absorption coefficient increasing from 108,000 to 148,000 +/- 2000 M-1.cm-1 (results given +/- S.E.M.; n = 3). The data provide evidence for a low-affinity site for bivalent metal ion binding in addition to the two constitutive calcium sites that are present in all plant peroxidases. The presence of a glutamic acid residue (Glu-141) at the entrance to the haem-binding pocket, analogous to Glu-146 in lignin peroxidase and not present in other plant peroxidases, may account for these novel properties.
authors.affiliation: Nitrogen Fixation Laboratory, John Innes Centre, Colney, Norwich, U.K.
authors.forename: I G
authors.initials: IG
authors.lastname: Gazarian
authors.forename: L M
authors.initials: LM
authors.lastname: Lagrimini
authors.forename: S J
authors.initials: SJ
authors.lastname: George
authors.forename: R N
authors.initials: RN
authors.lastname: Thorneley
mesh_terms: D001592:Benzyl Alcohols / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D006735:Horseradish Peroxidase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D018551:Solanum lycopersicum; D008274:Magnesium / Q000494:pharmacology; D008956:Models, Chemical; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013025:Glycine max; D014026:Nicotiana / Q000201:enzymology*
chemical_list: D001592:Benzyl Alcohols; D007527:Isoenzymes; D010940:Plant Proteins; D011994:Recombinant Proteins; D006735:Horseradish Peroxidase; D010544:Peroxidases; C042858:lignin peroxidase; C085547:tobacco anionic peroxidase; D008274:Magnesium; C042197:veratryl alcohol; D002118:Calcium