کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1191670 | 963575 | 2007 | 9 صفحه PDF | دانلود رایگان |
An intracellular peroxidase (POD) produced by genetically transformed root cultures of red beet (Beta vulgaris L.) was purified using a combination of (NH4)2SO4 fractionation and ion exchange chromatography resulting in 15-fold enhancement of activity. This enzyme exhibited highest activity (10,500 U mg−1 protein) and stability at pH 5.0 and retained over 70% of the activity for 20 min at 70 °C where horseradish peroxidase (HRP) – a vastly used commercial source, had lost its activity after 11 min. The purified enzyme showed highest preference for H2O2 as the substrate (Km value of 0.1). Among the H donors, the enzyme appeared to have affinity in the order of orthodianisidine > 2,2′-azino-bis(3-ethylbenz-thiazoline)-6-sulfonic acid > guaiacol. The purified POD was completely and competitively inhibited by periodate (HIO6-, Ki = 0.2 mM) whereas sodium azide (NaN3) was a non-competitive inhibitor. The purified POD had a molecular mass of 45 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and activity staining. This is the first report describing purification and characterization of POD from red beet hairy roots showing its better efficacy than commercial HRP.
Journal: Food Chemistry - Volume 105, Issue 3, 2007, Pages 1312–1320