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Gewählte Publikation:

Zamocky, M; Furtmüller, PG; Bellei, M; Battistuzzi, G; Stadlmann, J; Vlasits, J; Obinger, C; .
(2009): Intracellular catalase/peroxidase from the phytopathogenic rice blast fungus Magnaporthe grisea: expression analysis and biochemical characterization of the recombinant protein.
Biochem J. 2009; 418(2):443-451 FullText FullText_BOKU

Abstract:
Phytopathogenic fungi such as the rice blast fungus Magnaporthe grisea are unique in having two catalase/peroxidase (KatG) paralogues located either intracellularly (KatG1) or extracellularly (KatG2). The coding genes have recently been shown to derive from a lateral gene transfer from a (proteo)bacterial genome followed by gene duplication and diversification. Here we demonstrate that KatG1 is expressed constitutively in M. grisea. It is the first eukaryotic catalase/peroxidase to be expressed heterologously in Escherichia coli in high amounts, with high purity and with almost 100% haem occupancy. Recombinant MagKatG1 is an acidic, mainly homodimeric, oxidoreductase with a predominant five-co-ordinated high-spin haem b. At 25 degrees C and PI 17.0, the E(sic) (standard reduction potential) of the Fe(III)/Fe(II) Couple Was found to be -186 +/- 10 mV. It bound cyanide monophasically with an apparent bimolecular rate constant of (9.0 +/- 0.4) x 10(5) M (1) . s (1) at lit 17.0 and at 25 degrees C and with a K-d value of 1.5 mu M. Its predominantly catalase activity was characterized by a pH optimum at 6.0 and k(sic) and K-m values of 7010 s (1) and 4.8 mM respectively. In addition, it acts as a versatile peroxidase with a pH optimum in the range 5.0-5.5 using both one-electron [2,2'-,-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) o-dianisidine, pyrogallol or guaiacol] and two-electron (Br, I or ethanol) donors. Structure-Junction relationships are discussed with respect to data reported for prokaryotic KatGs, as is the physiological role of MagKatG1. Phylogenetic analysis suggests that (intracellular) MagKatG1 can be regarded as a typical representative for catalase/peroxidase of both phytopathogenic and saprotrophic fungi.
Autor*innen der BOKU Wien:
Furtmüller Paul Georg
Obinger Christian
Stadlmann Johannes
Zamocky Marcel
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Catalase/chemistry;Catalase/genetics*;Catalase/isolation & purification*;Catalase/physiology;Cloning, Molecular;Cyanides/metabolism;Enzyme Stability;Gene Expression Profiling;Gene Expression Regulation, Enzymologic;Gene Expression Regulation, Fungal;Hydrogen-Ion Concentration;Intracellular Space/enzymology;Magnaporthe/enzymology;Magnaporthe/genetics*;Magnaporthe/physiology;Models, Molecular;Oryza sativa/parasitology*;Peroxidases/chemistry;Peroxidases/genetics;Peroxidases/isolation & purification;Peroxidases/physiology;Recombinant Proteins/chemistry;Recombinant Proteins/genetics;Recombinant Proteins/isolation & purification;Structure-Activity Relationship;Thermodynamics;

Find related publications in this database (Keywords)
catalase
KatG
Magnaporthe grisea
oxidative stress
peroxidase
phytopathogenic fungus


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