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A spheroplast rate assay for determination of cell wall integrity in yeast
Author(s) -
Ovalle Rafael,
Lim Seung T.,
Holder Brigitte,
Jue Chong K.,
Moore Carol Wood,
Lipke Peter N.
Publication year - 1998
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19980930)14:13<1159::aid-yea317>3.0.co;2-3
Subject(s) - spheroplast , lysis , biology , saccharomyces cerevisiae , yeast , candida albicans , strain (injury) , cell wall , enzyme , tonicity , cell , microbiology and biotechnology , biochemistry , chromatography , escherichia coli , chemistry , gene , anatomy
The rate of formation of spheroplasts of yeast can be used as an assay to study the structural integrity of cell walls. Lysis can be measured spectrophotometrically in hypotonic solution in the presence of Zymolyase, a mixture of cell wall‐digesting enzymes. The optical density of the cell suspension decreases as the cells lyse. We optimized this assay with respect to enzyme concentration, temperature, pH, and growth conditions for several strains of Saccharomyces cerevisiae . The level of variability (standard deviation) was 1–5% between trials where the replications were performed on the same culture using enzyme prepared from the same lot, and 5–15% for different cultures of the same strain. This assay can quantitate differences in cell wall structure (1) between exponentially growing and stationary phase cells, (2) among different S. cerevisiae strains, (3) between S. cerevisiae and Candida albicans , (4) between parental and mutated lines, and (5) between drug‐ or chemically‐treated cells and controls. © 1998 John Wiley & Sons, Ltd.