Characterization of CYPllO from anabaena 7120

Anabaena 7120 is a fresh water photoantotrophic cyanobacterium. The chro mosome contains an 11kb excisable element with 9 open reading frances. One of the open reading frames encodes the cytochrome P450, CYP110. Most bacterial P450's are thought to aid in metabolizing xenobiotics for carbon and energy. To determine its biological role. Characterizaiton of CYP110 was undertaken. An E. coli expression system was constructed and CYP110 overrexpressed. Localization of CYP110 in E. coli was stadied using carbon monoxide difference spectra to monitor the protein. CYP110 associated with the mean brane and can be dissociated with high salt. Membrane associated bacterial P450's are often alkane inducible, Using a rabbit rabbit anti CYP110 antibody, in P450 cDNAs. Sequence comparison reveals that these house fly P450s are most closely related to mammalian mitochondrial P450s. CYPI2A1, abundantly represented in the cDNA library, is constitutively overexpressed in the insecticide-resistant strain Rutgers of the house fly. Overexpression is linked to a gene on chromosome II which is known to control insecticide resistance in this strain. CYP12A1 has been expressed in E. coli and Activity of CYP110 towards fatty acids is being examined using Na10₄-NADPH FNR, spinach ferredoxin and anaredoxin as electron donors.