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        Effect of Calcium on Immobilization of Rice (Oryza sativa L.) Peroxidase for Bioassays in Sodium Alginate and Agarose Gel

        Kavita Shah,Sareeta Nahakpam,Puneeta Singh 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.5

        The direct immobilization of soluble peroxidase isolated and partially purified from shoots of rice seedlings in calcium algi-nate beads and in calcium agarose gel was carried out. Peroxidase was assayed for guaiacol oxidation products in pres-ence of hydrogen peroxide. The maximum specific activity and immobilization yield of the calcium agarose immobilized per-oxidase reached 2,200 U mg-¹ protein (540 mU cm-³ gel) and 82%, respectively. In calcium alginate the maximum activity of peroxidase upon immobilization was 210 mU g-¹bead with 46% yield. The optimal pH for agarose immobilized peroxidase was 7.0 which differed from the pH 6.0 for soluble peroxidase. The optimum temperature for the agarose immobilized per-oxidase however was 30°C, which was similar to that of soluble peroxidase. The thermal stability of calcium agarose immo-bilized peroxidase significantly enhanced over a temperature range of 30~60°C upon immobilization. The operational stabil-ity of peroxidase was examined with repeated hydrogen peroxide oxidation at varying time intervals. Based on 50% conver-sion of hydrogen peroxide and four times reuse of immobilized gel, the specific degradation of guaiacol for the agarose im-mobilized peroxidase increased three folds compared to that of soluble peroxidase. Nearly 165% increase in the enzyme protein binding to agarose in presence of calcium was noted. The results suggest that the presence of calcium, ions help in the immobilization process of peroxidase from rice shoots and mediates the direct binding of the enzyme to the agarose gel and that agarose seems to be a better immobilization matrix for peroxidase compared to sodium alginate.

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