Abstract:
Cytochromes P450 are powerful enzymes capable of catalyzing oxidation of unactivated carbon-hydrogen bonds at physiological temperatures and pH. They are ubiquitous to life, yet the way in which their sequence dictates substrate selection and specificity is poorly understood. To investigate the problem of how sequence dictates substrate specificity, I focused on two Pseudomonas strains isolated from pine forest soil capable of subsisting on α-terpineol, (R)-(+)-limonene, and (1R)-(+)-α-pinene. Volatile metabolites show transformations of these carbon sources characteristic of cytochromes P450, and a cytochrome P450 was purified from one of the two organisms when it was grown with α-terpineol as its sole carbon source. I collected and analyzed transcriptomes from the two organisms grown with and without the terpenes to identify sequences which are expressed only in the presence of monoterpenes. While sequence data from the organism the cytochrome P450 was isolated from is too short to be conclusive, the other organism contains a sequence which shows promise as worthwhile of further investigation.
