A comparison of quantitative and qualitative superoxide dismutase assays for application to low temperature microalgae

P.J. Janknegt, J.W. Rijstenbil, W.H. van de Poll, T.S. Gechev, A.G.J. Buma

    Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

    15 Downloads (Pure)


    Antioxidant enzymes such as superoxide dismutase (SOD) play a key role in the removal of reactive oxygen species produced during visible and ultraviolet irradiance stress in microalgae and plants. However, little is known about the enzymatic antioxidative stress responses in ecologically important Antarctic marine microalgae. SOD in particular is difficult to analyze, possibly due to problems in obtaining sufficient quantities necessary for reliable and reproducible enzymatic assays. The aim of the present work was to create a sensitive, easy-to-use and reliable method for SOD determination in Antarctic microalgal material by comparing and optimizing existing protein extraction procedures and SOD assays in the marine Antarctic diatom Chaetoceros brevis. Optimization was achieved in cell disruption (sonication) and protein extraction procedures, extraction buffers, SOD assay methods (xanthine/xanthine oxidase and NBT/riboflavin photometric quantitative methods and native gel electrophoresis qualitative method) and the assay temperature. Protein extraction was optimal at low sonication amplitudes after a few pulses, irrespective of the type of buffer used. Extraction efficiency varied highly between the tested buffers; most protein was extracted in the presence of 1% of Triton X-100. SOD activity was best quantified using the NBT/riboflavin method in combination with a buffer containing potassium phosphate and Triton X-100. Moreover, the NBT/riboflavin method was demonstrated to be the most reliable and sensitive method at low temperatures (5 °C).
    Originele taal-2Engels
    Pagina's (van-tot)218-226
    TijdschriftJournal of Photochemistry and Photobiology B: Biology
    Nummer van het tijdschrift3
    StatusGepubliceerd - 2007


    Duik in de onderzoeksthema's van 'A comparison of quantitative and qualitative superoxide dismutase assays for application to low temperature microalgae'. Samen vormen ze een unieke vingerafdruk.

    Citeer dit