Partial purification of NADH:menadione oxidoreductase from S. wolfei

Purification stepProtein amt (mg)Sp act (U/mg)aYield (%)Purification factor
NADH oxidationbNADH:menadione oxidoreductasec
First preparation run
    Cell extract63.90.31.7NAdNA
    Soluble fraction41.40.33.41001
    DEAE-Sepharose CL-4B (pH 8.0)10.20.569.9722.9
    MonoQ (pH 8.0)1.072.4919.9155.8
    POROS HQ/H (pH 8.0)0.61.6516.574.8
    MonoQ (pH 7.5)0.042.3459.31.617.4
Second preparation run
    Cell extract721.75.9NANA
    Soluble fraction45.43.04.4NANA
    Membrane fraction28.80.31.551001
    Solubilized membrane protein14.160.375.141633.3
    DEAE-Sepharose CL-6B (pH 8.0)7.920.24.7584.33.06
    MonoQ (pH 8.0)0.881.4620.239.813.03
    MonoQ (pH 7.5)0.480.6626.4612.717.07
  • a Mean activities of three independent measurements are shown.

  • b Unspecific oxidation of NADH before the addition of menadione.

  • c Values corrected for unspecific NADH-oxidizing activity.

  • d NA, not applicable.