title: In situ assay of intracellular enzymes of yeast (Kluyveromyces fragilis) by digitonin permeabilization of cell membrane.
abstract: 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.
authors.affiliation: Department of Food Chemistry, Central Food Technological Research Institute, Mysore, India.
authors.forename: L R
authors.initials: LR
authors.lastname: Gowda
authors.forename: M S
authors.initials: MS
authors.lastname: Joshi
authors.forename: S G
authors.initials: SG
authors.lastname: Bhat
mesh_terms: D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D000642:Ammonia-Lyases / Q000378:metabolism*; D001220:Aspartate Ammonia-Lyase / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002474:Cell-Free System; D004072:Digitonin; D005696:Galactosidases / Q000378:metabolism*; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism*; D007700:Kinetics; D007716:Kluyveromyces / Q000201:enzymology*; D004718:Saccharomycetales / Q000201:enzymology*; D001616:beta-Galactosidase / Q000378:metabolism*
chemical_list: D000426:Alcohol Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D005696:Galactosidases; D001616:beta-Galactosidase; D000642:Ammonia-Lyases; D001220:Aspartate Ammonia-Lyase; D004072:Digitonin