Purification and catalytic properties of two manganese peroxidase isoenzymes from Pleurotus eryngii.
The ligninolytic basidiomycetes Pleurotus eryngii, Pleurotus ostreatus, Pleurotus pulmonarius and Pleurotus sajor-caju did not exhibit detectable levels of manganese peroxidase (MP) when grown in liquid media with ammonium tartrate as N source. However, after examination of cells grown on different organic N-based media, high MP activity was obtained in peptone medium, up to nearly 3 U/ml in cultures of P. eryngii. Moreover, Mn2+ supplementation was not used to produce MP, since all Mn2+ concentrations assayed (1-4000 microM) inhibited production of this enzyme in liquid medium. Two MP isoenzymes were purified to homogeneity from shaken or stationary cultures of P. eryngii grown in peptone medium. The purification process (which included chromatography on Biorad Q-cartridge, Sephacryl S-200 and Mono-Q) attained 56% activity yield with a purification factor of 25. The isoenzymes differed in pI (3.75 and 3.65), N-terminal sequence and some catalytic properties. They were in some aspects (e.g, molecular mass of 43 kDa) similar to Phanerochaete chrysosporium MP but exhibited some distinct characteristics, including Mn(2+)-independent peroxidase activities against 2,6-dimethoxyphenol and veratryl alcohol, and higher resistance to H2O2. Recent studies have shown that MP are ubiquitous enzymes in ligninolytic fungi, but the results obtained suggest that differences in catalytic properties probably exist between different Mn(2+)-oxidizing peroxidases produced by these fungi.
Martínez
M J
MJ
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Ruiz-Dueñas
F J
FJ
Guillén
F
F
Martínez
A T
AT
D000595:Amino Acid Sequence; D002384:Catalysis; D003470:Culture Media; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010544:Peroxidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011124:Polyporaceae / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
D003470:Culture Media; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008345:Manganese; D006861:Hydrogen Peroxide; D010544:Peroxidases; C051129:manganese peroxidase