A simple colorimetric determination of the manganese content in photosynthetic membranes |
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Authors: | Boris K Semin Michael Seibert |
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Institution: | (1) Department of Biophysics, Faculty of Biology, Lomonosov Moscow State University, 119991 Moscow, Russia;(2) Energy Sciences Directorate, National Renewable Energy Laboratory, Golden, Colorado 80401, USA |
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Abstract: | The functional Mn content of intact photosystem II membrane fragments was measured as 4.06 ± 0.13 Mn/reaction center when
determined using a simple, sensitive colorimetric assay that will also work with thylakoids and core complexes. This procedure
requires minimal sample material, does not need expensive assay equipment, requires four simple steps, and only takes 20–30 min
to perform. These include (a) removal of the adventitious Mn ions by CaCl2 treatment of the membranes, (b) extraction of the Mn from the O2-evolving complex with hydrochloric acid, (c) purification of the extract by centrifugation followed by filtration of the
supernatant through an Acrodisc syringe filter (0.2 μm nylon membrane), and (d) colorimetric determination of Mn in the extract
using the reaction of the chromogenic agent, 3,3′,5,5′-tetramethylbenzidine, with previously oxidized Mn(II) cations carried
out at high pH. The colorimetric assay itself has been used previously by Serrat (Mikrochim Acta 129:77–80, 1998) for assaying
Mn concentrations in sea water and drinking water. |
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Keywords: | Manganese assay Colorimetric assay Manganese content Photosystem II Oxygen-evolving complex |
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