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Table 1 Specific activity of recombinant F6H proteinsa

From: Molecular characterization and functional expression of flavonol 6-hydroxylase

Step Specific Activity (pkat/mg)
rF6Hb
   Cell Lysate 0
   Ni-NTA 0.091
   Ni-NTA (30°C) 0.087
   Ni-NTA + Superose 12 0.103
   Superose 12 0.075
   Cleavage of (His)6-tag after Ni-NTA 0.044
rF6Hy (intracellular)
   Cell Lysate 0
   Ni-NTA 0.17
   Ni-NTA + Superdex 75 0.21
   Superdex 75 0.087
rF6Hys (secreted)
   (NH4)2SO4 (35–70%) 0
   (NH4)2SO4 + Ni-NTA 0.11
   (NH4)2SO4 + Ni-NTA + Superdex 75 0.08
   Superdex 75 0.04
  1. aAll assays were carried in the presence of assay buffer, 5 μM 3,7,4'-trimethylquercetin and ODD cofactors using the F6H enzyme assay as described in [3].