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Localization of Alkaline Phosphatase in Yeast

Abstract

The Gomori technique1 for alkaline phosphatase localization has been extensively employed with animal and plant material ; but it does not appear to have been employed with yeasts. The presence of acid and alkaline phosphatases in yeasts is Well known2,3,4; but the location of these enzymes in the cell has not previously been described. We have applied the Gomori technique to preparations of Saccharomyces cerevisie (strain ‘Yeast foam', Carls-berg No. 237). Cells were grown in pure culture in 10 ml. of 8 per cent Pilsner Wort in Freudenreich flasks at 25° C. for 24 hr. At the end of this time the medium was poured off and 10 ml. of sterilized distilled water was added to the cells sedimented at the bottom of the flask. The cells were suspended in the water by shaking, and drops of the suspension placed on microscope slides. The cells were allowed to dry in air for 10-30 min. and then incubated in one of the several phosphate compound substrates in a veronal buffer solution containing magnesium and calcium at pH 9·2 for 16-40 hr. at 37° C. The cobalt sulphide method of staining1 was then applied.

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NICKERSON, W., KRUGELIS, E. & ANDRÈSEN, N. Localization of Alkaline Phosphatase in Yeast. Nature 162, 192–193 (1948). https://doi.org/10.1038/162192a0

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