Characterization of structure and activity of garlic peroxidase (POX(1B)).
Structural characterization and study of the activity of new POX(1B) protein from garlic which has a high peroxidase activity and can be used as a biosensor for the detection of hydrogen peroxide and phenolic compounds were performed and compared with the findings for other heme peroxidases. The structure-function relationship was investigated by analysis of the spectroscopic properties and correlated to the structure determined by a new generation of high-performance hybrid mass spectrometers. The reactivity of the enzyme was analyzed by studies of the redox activity toward various ligands and the reactivity with various substrates. We demonstrated that, in the case of garlic peroxidase, the heme group is pentacoordinated, and has an histidine as a proximal ligand. POX(1B) exhibited a high affinity for hydrogen peroxide as well as various reducing cosubstrates. In addition, high enzyme specificity was demonstrated. The k(cat) and K(M) values were 411 and 400 mM(-1) s(-1) for 3,3',5,5'-tetramethylbenzidine and 2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), respectively. Furthermore, the reduction of nitro compounds in the presence of POX(1B) was demonstrated by iron(II) nitrosoalkane complex assay. In addition, POX(1B) showed a great potential for application for drug metabolism since its ability to react with 1-nitrohexane in the presence of sodium dithionite was demonstrated by the appearance of a characteristic Soret band at 411 nm. The high catalytic efficiency obtained in the case of the new garlic peroxidase (POX(1B)) is suitable for the monitoring of different analytes and biocatalysis.
Equipe de Chimie Bioorganique et Bioinorganique, Institut de Chimie Moléculaire et Matériaux d'Orsay, UMR 8182, CNRS, Université Paris-Sud, Orsay, France.
Sarra
S
El Ichi
Anna
A
Miodek
Hélène
H
Sauriat-Dorizon
Jean-Pierre
JP
Mahy
Céline
C
Henry
Mohamed Nejib
MN
Marzouki
Hafsa
H
Korri-Youssoufi
D055162:Biocatalysis; D015374:Biosensing Techniques; D004789:Enzyme Activation; D005737:Garlic / Q000201:enzymology*; D006861:Hydrogen Peroxide / Q000032:analysis; D009195:Peroxidase / Q000737:chemistry* / Q000378:metabolism*; D010636:Phenols / Q000032:analysis; D011487:Protein Conformation
D010636:Phenols; D006861:Hydrogen Peroxide; D009195:Peroxidase