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Table 1 Purification of 2,5-DKCMO via nickel-based affinity chromatography and imidazole removal

From: Recombinant expression and purification of the 2,5-diketocamphane 1,2-monooxygenase from the camphor metabolizing Pseudomonas putida strain NCIMB 10007

Step V [mL] Volumetric activity a [U/mL] Activity a [U] Protein amount b [mg/mL] Specific activity [mU/mg] Yield [%] Factor
Crude extract 30 0.0021 0.063 14 0.15 100 1
Purified and desalted 7 0.0015 0.0103 1.6 0.90 16 6
  1. aActivity was determined towards (+)-camphor and analyzed by GC-MS.
  2. Activity of the purified protein containing imidazole prior to the size exclusion chromatography could not be determined, since imidazole interferes with the used GC-MS column.
  3. bProtein amount as determined by the BCA assay.