Mar 2019 DOI 10.14302/issn.2471-2140.jaa-19-2699
M.M. Masoud HassanCorresponding author
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Xanthine oxidase is a commercially important enzyme with wide area of medical applications to develop diagnostic kits. Xanthine oxidase was extracted, purified and characterized from sheep liver (SLXO). The purification procedure involved acetone precipitation and chromatography on DEAE-cellulose and Sephacryl S-300 columns. The sheep liver xanthine oxidase was homogeneously purified 31.8 folds with 3.5 U/mg specific activity and 24.1% recovery. SLXO native molecular weight was 150 kDa and on SDS-PAGE appeared as single major band of 75 kDa representing a homodimer protein. Isoelectric focusing of the purified SLXO resolved into two closely related isoforms with pI values of 5.6 and 5.8. The apparent Km for xanthine oxidase at optimum pH 7.6 was found to be 0.9 mM xanthine. FeCl2 and NiCl2 increased the activity of SLXO, while CuCl2 and ZnCl2 were found to be potent inhibitors of the purified enzyme. Allopurinol inhibits SLXO competitively with one binding site on the purified molecule and Ki value of 0.06 mM.
Dec 2015 DOI 10.14302/issn.2471-2140.jaa-15-715
Kevers ClaireCorresponding author
Plant Molecular Biology and Biotechnology and CEDEVIT B22, University of Liège Chemin de la Vallée, 4, Sart Tilman, 4000 Liège Belgium
Since a few years, more and more attention has been specifically given to dietary antioxidants as agents promoting health and preventing the incidence of diseases. As part of these efforts, analytical methods and assays have been developed to measure the antioxidant content in food substances. In this paper, the antioxidant capacity of 17 orange juices is determined by various assays (DPPH, ORAC, heamolysis, xanthine/xanthine oxidase) as the content in ascorbic acid and total phenolics. The results evidence all the complexity to evaluate the in vitro antioxidant capacity of foods. In very general terms, in spite of the wide utilization in these tests (FRAP, TAC, ORAC TRAP and others), their significance remains obscure. The discrepancy of the results and the absence of good correlation between the assays clearly highlight all the importance of understanding the strengths and weakness of assays evaluating antioxidant potential of a food at the risk of giving erroneous information to the consumer. It is clear that the use of "total antioxidant capacity" assays for the in vitro assessment of antioxidant quality of food does not be employed by food industrials as a marketing argument or for the assessment of the "wholesomeness" of a food.