| Pseudomonas aeruginosa is a pathogenic bacterium that is deadly to patients who suffer from Cystic Fibrosis. To develop a drug to stop the P. aeruginosa infection, the heme oxygenase reaction must be better understood. Heme oxygenases from Neisseriae meningitides (nm-HO) and Pseudomonas aeruginosa (pa-HO) share significant sequence identity (37%). N m-HO produces biliverdin IXalpha, whereas pa-HO yields predominantly biliverdin IXdelta due to an in-plane heme rotation of 110°. To investigate the in-plane conformation of the heme a chimeric protein was made. It was found that a double mutation in the chimeric protein stabilizes the in-plane conformer conducive to delta-meso hydroxylation.; The electron transfer process for the heme oxygenase reaction is known to be different from that of human heme oxygenase. The ferredoxin (Bfd) and ferredoxin reductase (FPR) were unknown until recently, however; they are the enzymes responsible for the seven electrons required for the degradation of heme. The expression and purification method were optimized for FPR, and the cofactor was determined. |
