New method for detection of enzymatic activity of flavocytochrome b2 in electrophoregrams |
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Authors: | Gaĭda G Z Stel'mashchuk S Ia Smutok O V Gonchar M V |
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Affiliation: | Institute of Cell Biology, Ukrainian Academy of Sciences, Lviv, 79005 Ukraine. |
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Abstract: | A new method of visualization of the activity of flavocytochrome b2 (FC b2; L-lactate: ferricytochrome c oxidoreductase, EC 1.1.2.3) in electrophoretograms was developed, based on the interaction between ferrocyanide (generated during the enzymatic reaction) and Fe23+, resulting in the formation of intensely colored precipitates of Berlin blue. The main advantages of this method were its high sensitivity (less than 0.005 U FC b2 was detected within a suitable time period) and the stability of the dye formed. The method developed can be used for determining FC b2 activity in cell-free extracts (e.g., in the selection of FC b2 producers) and monitoring chromatographic purification of proteins, as well as in other cases associated with FC b2 assessment. |
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