Cloning and gene expression of cytochrome P450 gene from Alcanivorax borkumensis Bacterium

Alcanivorax borkumensis is a marine bacterium that has ability to grow on limited substrates that mainly is alkanes. The ability to use wide range of hydrocarbons is advantage of this bacterium to other marine community bacteria. A. borkumensis have two genetic systems for alkane biodegradation. The First system is alkane hydroxylase (alk-B1and alk-B2) and the second system is  cytochrome P450. Until now there is not any report on cloning of cytochrome P450 gene of A. borkumensis. In this study cytochrome P450 gene from A. borkumensis was cloned. At first P450 gene was cloned in pBluescript plasmid by blunt cloning and insertion was confirmed by colony PCR.  Then P450 gene was cloned in PET-26 expression vector and finally, IPTG induced gene expression in E.coli BL-21 bacterium. It was shown that longer induction time led to more expression level. The recombinant enzyme has difference absorbance in 450 nm that confirmed the activity of this recombinant enzyme.