CYTOCHEMICAL LOCALIZATION OF DEHYDROGENASE AND DIAPHORASE ACTIVITY IN BACILLUS MEGATERIUM

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Abstract
Using 3-(4,5-dimethyl-thiazolyl-2) 2,5-diphenyltetrazolium bromide and cobalt chelation of its formazan, the localization of succinic, [beta]-hydroxybutyric dehydrogenat and diphosphopyrldine nucleotide-diaphorase was studied in B. megaterium. Formazan pigment showed as dots of 0.1 to 0.2 [mu] diameter at regular intervals within the cytoplasm and often was disposed as parts of a spiral. During protoplast formation, the dye deposits adhered to the cytoplasmic body. On stabilized protoplasts, enzymatically formed pigment accumulated in the region of the cytoplasmic membrane. Quantitative studies showed tetrazolium reduction by B. megaterium to be substrate dependent. Highest activity was observed during the early phase of growth and with succinate as a substrate. The activity of protoplasts was less than the activity of intact cells.