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

AMB Express

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

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

^aActivity was determined towards (+)-camphor and analyzed by GC-MS.
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.
^bProtein amount as determined by the BCA assay.