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1 Biotechnologia

1 2002

1 Salek A. T.

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Classic techniques for improvement of industrial yeast strains: Part III ? A method for enucleation of Saccharomyces sp.

100%

Salek A. T.

Biotechnologia

2002

nr 1

187-194

The first method for enucleation of yeast Saccharomyces cerevisiae is reported. Various strains, including some killer strain and respiratory-deficient mutants of Saccharomyces cerevisiae were enucleated after treatment with cytochalasin B. Removal of nuclei from protruding sphaeroplasts was induced by centrifugation in a Percoll density gradient. The enucleation yield (which averaged about 80%) and the quality of the cytoplasts were best when the yeast culture had been synchronized with nocodazole before the preparation. The presence of 1 mM CaCl2 and ATP (10 muM) in the enucleation medium prevented the formation of fragile products or aggregation. Cytoplasts could be stored for at least 1 day without visible deterioration.