TABLE 2 

Purification of the PDH complex of E. faecalis expressed in S. cerevisiaea

SamplePurification stepSp act (μmol/min/mg of protein)Purification factor
Homogenate0.067 ± 0.0031
Cytosol C1Centrifugation0.062 ± 0.0010.9
Cytosol C2Centrifugation0.071 ± 0.0031.1
Cytosol C3Concentration on 100-kDa cutoff filter0.083 ± 0.0031.2
Column fraction 14Column separation8.83 ± 0.25131
  • a PDH activity was measured in homogenates of strain IMY104. In the next step, mitochondrial and cytosolic fractions of the homogenate were separated. The cytosolic fraction was further purified and applied onto a size exclusion chromatographic column. Fraction 14 obtained from the column showed the highest specific activity of the PDH complex. Means and standard deviations were obtained from duplicate measurements on a single column. An independent duplicate experiment gave similar results.