Purification and characterization of manganese peroxidase of the white-rot fungus Irpex lacteus.
The production of manganese peroxidase (MnP) by Irpex lacteus, purified to electrophoretic homogeneity by acetone precipitation, HiPrep Q and HiPrep Sephacryl S-200 chromatography, was shown to correlate with the decolorization of textile industry wastewater. The MnP was purified 11.0-fold, with an overall yield of 24.3%. The molecular mass of the native enzyme, as determined by gel filtration chromatography, was about 53 kDa. The enzyme was shown to have a molecular mass of 53.2 and 38.3 kDa on SDS-PAGE and MALDI-TOF mass spectrometry, respectively, and an isoelectric point of about 3.7. The enzyme was optimally active at pH 6.0 and between 30 and 40 degrees C. The enzyme efficiently catalyzed the decolorization of various artificial dyes and oxidized Mn (II) to Mn (III) in the presence of H(2)O(2). The absorption spectrum of the enzyme exhibited maxima at 407, 500, and 640 nm. The amino acid sequence of the three tryptic peptides was analyzed by ESI Q-TOF MS/MS spectrometry, and showed low similarity to those of the extracellular peroxidases of other white-rot basidiomycetes.
Shin
Kwang-Soo
KS
Department of Microbiology, Daejeon University, Republic of Korea. shinks@dju.ac.kr
Kim
Young Hwan
YH
Lim
Jong-Soon
JS
D001487:Basidiomycota / Q000201:enzymology*; D002853:Chromatography, Liquid; D004396:Coloring Agents / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005590:Fractional Precipitation; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013057:Spectrum Analysis; D013696:Temperature; D014874:Water Pollutants, Chemical / Q000378:metabolism
D004396:Coloring Agents; D014874:Water Pollutants, Chemical; D008345:Manganese; D006861:Hydrogen Peroxide; D010544:Peroxidases; C051129:manganese peroxidase