A new thermostable peroxidase from garlic Allium sativum: purification, biochemical properties, immobilization, and use in H2O2 detection in milk.

Abstract:

:Analysis of peroxidase activity by native polyacrylamide gel electrophoresis (PAGE) from a garlic bulb (Allium sativum L) extract showed two major activities (designated POX1 and POX2). The POX2 isoenzyme was purified to homogeneity by ammonium sulfate precipitation, gel filtration, and cation-exchange chromatography. The purified enzyme was found to be monomeric with a molecular mass of 36.5 kDa, as determined by sodium dodecyl sulfate-PAGE. The optimum temperature ranged from 25 to 40 degrees C and optimum pH was about 5.0. The apparent Km values for guaiacol and H2O2 were 9.5 and 2 mM, respectively. POX2 appeared highly stable since 50% of its activity was conserved at 50 degrees C for 5 h. Moreover POX2 was stable over a pH range of 3.5-11.0. Immobilization of POX2 was achieved by covalent binding of the enzyme to an epoxy-Sepharose matrix. The immobilized enzyme showed great stability toward heat and storage when compared with soluble enzyme. These properties permit the use of this enzyme as a biosensor to detect H2O2 in some food components such as milk or its derivatives.

authors

Marzouki SM,Limam F,Smaali MI,Ulber R,Marzouki MN

doi

10.1385/abab:127:3:201

subject

Has Abstract

pub_date

2005-12-01 00:00:00

pages

201-14

issue

3

eissn

0273-2289

issn

1559-0291

pii

ABAB:127:3:201

journal_volume

127

pub_type

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