✦ LIBER ✦

In situ assay of intracellular enzymes of yeast (Kluyveromyces fragilis) by digitonin permeabilization of cell membrane

✍ Scribed by L.R. Gowda; M.S. Joshi; S.G. Bhat

Publisher: Elsevier Science
Year: 1988
Tongue: English
Weight: 439 KB
Volume: 175
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(88)90579-9

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✦ Synopsis

The yeast, Kluyveromyces fragilis was permeabilized to a number of low-molecular-weight substrates using digitonin. The activities of intracellular yeast enzymes, viz., alcohol dehydrogenase (ADH), beta-galactosidase, glucose-6-phosphate dehydrogenase, aspartase, and hexokinase were found to be much higher in the permeabilized cells than the untreated cells. The optimum conditions for permeabilization with reference to ADH were 0.1% digitonin at 37 degrees C for 15 min. The ADH activity in permeabilized cells was several-fold higher than that in cell free extracts prepared by either physical or chemical methods.

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