[New method for detection of enzymatic activity of flavocytochrome b2 in electrophoregrams].
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.
Institute of Cell Biology, Ukrainian Academy of Sciences, Lviv, 79005 Ukraine.
G Z
GZ
Gaĭda
S Ia
SIa
Stel'mashchuk
O V
OV
Smutok
M V
MV
Gonchar
D004586:Electrophoresis / Q000379:methods; D005290:Ferric Compounds; D005295:Ferrocyanides / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000032:analysis* / Q000378:metabolism; D042844:L-Lactate Dehydrogenase (Cytochrome); D010843:Pichia / Q000201:enzymology; D012680:Sensitivity and Specificity
D005290:Ferric Compounds; D005295:Ferrocyanides; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); C020354:hexacyanoferrate II; C000170:ferric ferrocyanide