Novel yeast cell dehydrogenase activity assay in situ

• Autorzy: Berlowska J , Kregiel D. , Klimek L. , Orzeszyna B. , Ambroziak W.
• Języki publikacji: EN

Abstrakty

EN

The aim of this research was to develop a suitable method of succinate dehydrogenase activity assay in situ for different industrial yeast strains. For this purpose different compounds: EDTA, Triton X-100, sodium deoxycholate, digitonin, nystatin and β-mercaptoethanol were used. The permeabilization process was controlled microscopically by primuline staining. Enzyme assay was conducted in whole yeast cells with Na-succinate as substrate, phenazine methosulfate (PMS) as electron carrier and in the presence one of two different tetrazolium salts: tetrazolium blue chloride (BT) or cyanoditolyl tetrazolium chloride (CTC) reduced during the assay. In comparabile studies of yeast vitality the amount of intracellular ATP was determined according to luciferin/luciferase method. During the succinate dehydrogenase assay in intact yeast cells without permeabilization, BT formazans were partially visualized in the cells, but CTC formazans appeared to be totally extracellular or associated with the plasma membrane. Under these conditions there was no linear relationship between formazan color intensity signal and yeast cell density. From all chemical compounds tested, only digitonin was effective in membrane permeabilization without negative influence on cell morphology. Furthermore, with digitonin-treated cells a linear relationship between formazan color intensity signal and yeast cell number was noticed. Significant decreasing of succinate dehydrogenase activity and ATP content were observed during aging of the tested yeast strains.

Słowa kluczowe

EN

yeast; succinate dehydrogenase; living cell; microorganism; permeabilization; dehydrogenase activity; yeast cell

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2006
• Tom: 55
• Numer: 2
• Opis fizyczny: p.127-131,fig.,ref.

Twórcy

autor

Berlowska J

• Technical University of Lodz, Wolczanska 171/173, 90-924 Lodz, Poland

autor

Kregiel D.

autor

Klimek L.

autor

Orzeszyna B.

autor

Ambroziak W.

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