[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.
Gaĭda
G Z
GZ
Institute of Cell Biology, Ukrainian Academy of Sciences, Lviv, 79005 Ukraine.
Stel'mashchuk
S Ia
SIa
Smutok
O V
OV
Gonchar
M V
MV
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