Purification and characterization of an intracellular catalase-peroxidase from Penicillium simplicissimum.
The first dimeric catalase-peroxidase of eucaryotic origin, an intracellular hydroperoxidase from Penicillium simplicissimum which exhibited both catalase and peroxidase activities, has been isolated. The enzyme has an apparent molecular mass of about 170 kDa and is composed of two identical subunits. The purified protein has a pH optimum for catalase activity at 6.4 and for peroxidase at 5.4. Both activities are inhibited by cyanide and azide whereas 3-amino-1,2,4-triazole has no effect. 3,3'-Diaminobenzidine, 3,3'-dimethoxybenzidine, guaiacol, 2,6-dimethoxyphenol and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) all serve as substrates. The optical spectrum of the purified enzyme shows a Soret band at 407 nm. Reduction by dithionite results in the disappearance of the Soret band and formation of three absorption maxima at 440, 562 and 595 nm. The prosthetic group was identified as a protoheme IX and EPR spectroscopy revealed the presence of a histidine residue as proximal ligand. In addition to the catalase-peroxidase, an atypical catalase which is active over a broad pH range was also partially purified from P. simplicissimum. This catalase is located in the periplasm and contains a chlorin-type heme as prosthetic group.
Fraaije
M W
MW
Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
Roubroeks
H P
HP
Hagen
W R
WR
Van Berkel
W J
WJ
D001386:Azides / Q000494:pharmacology; D002374:Catalase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D003486:Cyanides / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007424:Intracellular Fluid / Q000201:enzymology; D008024:Ligands; D015394:Molecular Structure; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D011166:Porphyrins / Q000737:chemistry; D011487:Protein Conformation; D013053:Spectrophotometry; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity
D001386:Azides; D003486:Cyanides; D008024:Ligands; D011166:Porphyrins; C006969:chlorin; D006418:Heme; D002374:Catalase; D009195:Peroxidase