DETECTION OF DIVERSE CYTOCHROME P450-DEPENDENT BIOOXYGENATION CATALYSTS IN MICROORGANISMS USING A MULTIPURPOSE INDUCER

Screening for new microbial cytochromes P450 to create a useful pool o f oxygenating biocatalysts can be facilitated by applying multivalent inducers for it. The broad inducer spectrum for cytochrome P450 of Aci netobacter calcoaceticus EB 104 has been analyzed as a model for struc ture-function relationships governing the inducer recognition of hydro carbon-oxidizing microbial cytochromes P450 with the aim to deduce str uctural requirements for the desired multivalent inducer. The cytochro me was induced by all compounds of adequate hydrophobicity that contai n coplanar structural elements. It was concluded that due to the inher ent low specificity of hydrophobic binding forces some few simple-stru ctured hydrocarbons with a sterically flexible backbone can induce a v ariety of different hydrocarbon-oxidizing microbial cytochromes P450. This approach has been proved to be successfull by induction of cytoch rome P450 with the help of n-hexane in Bacillus megaterium, in various strains of the genus Rhodococcus and in the yeast Candida apicola. Cy tochrome P450 of Rhodococcus rhodochrous IMET 7278, that has partially been purified from n-hexane-induced cells, exhibited an induction pro file differing to P450 of Acinetobacter.