Purification of anthranilate-CoA ligase

Purification stepTotal activity (U)Amt of total protein (mg)Sp act (U/mg protein)Yield (%)Purification (fold)
Crude extract93.4a4,1920.022a1001
50% ammonium sulfate pellet88.78200.108954.9
His-Select fraction (Ni2+ affinity)61.8800.7736635
Butyl-Sepharose fraction21.4211.022346
HiTrap Q fraction15.2121.271658
  • a Enzymatic activity values for the crude extract are extrapolated from the sum of the total activities in the pellet and the supernatant from the 50% ammonium sulfate precipitation. Interfering activities gave artificially low values when the activity in the crude extract was measured directly.