Optimisation Of Expression And Immobilized Metal Ion Affinity Chromatographic Purification Of Recombinant (His)(6)-Tagged Cytochrome P450 Hydroperoxide Lyase In Escherichia Coli

Fatty acid hydroperoxide lyase (HPL) is a cytochrome P450 acting on fatty acid’s hydroperoxides in many plants. The optimisation of the expression of recombinant (His) -tagged HPL in Escherichia coli is described: the highest HPL 6 production yield were obtained with TB medium supplemented with 2.5 mM d-aminolevulinic acid and 0.5 mM IPTG. For the first time, the time course expression of a plant P450 in a bench-scale fermentor is detailed and the amount of recombinant HPL production is 16.3 mg/ l. The UV–Visible spectrum of the recombinant (His) -tagged HPL have been recorded after a Ni -based affinity chromatography (IMAC).