Total and specific activity of DAP in C. albicans ATCC 10231 (membrane fraction)
Cellular fractions were prepared from cells growing in the corresponding medium. Biomass from the culture medium was recovered by centrifugation at 5 °C (2000g), washed twice with 0.1 M Tris buffer, pH 7.0, and disrupted with glass beads to produce a cell homogenate. This fraction was centrifuged at 23,000g, and the resulting supernatant was centrifuged at 100,000g to separate the soluble fraction (cytoplasmic) from the membrane fraction. The 100,000g fraction (membranes) was used to determine DAP enzymatic activity against Ala-pro-pNA as substrate.
Total activity (mU ml−1).
Specific activity (mU mg protein−1).
Total and specific activity of DAP in C. albicans ATCC 10231 (membrane fraction)
Cellular fractions were prepared from cells growing in the corresponding medium. Biomass from the culture medium was recovered by centrifugation at 5 °C (2000g), washed twice with 0.1 M Tris buffer, pH 7.0, and disrupted with glass beads to produce a cell homogenate. This fraction was centrifuged at 23,000g, and the resulting supernatant was centrifuged at 100,000g to separate the soluble fraction (cytoplasmic) from the membrane fraction. The 100,000g fraction (membranes) was used to determine DAP enzymatic activity against Ala-pro-pNA as substrate.
Total activity (mU ml−1).
Specific activity (mU mg protein−1).
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