Cytochrome P450 enzymes are part of an extremely large family that can catalyse many different types of chemical reactions, including oxidations as well as reductions. P450 enzymes are able to introduce – region and enantio selective – an oxygen atom onto an activated or unactivated carbon or a heteroatom (like nitrogen, oxygen or sulphur) in a substrate. This type of reaction is very hard or even not accomplishable through conventional chemical synthesis.
P450 enzymes could be ideal tools for the organic chemist if there is a technology available to create and find P450 enzymes with the desired catalytic characteristics. (designer biocatalysts)
Bacterial P450s are with activities as high as 4600 mol-1 min-1 mol-1 for P450 BM-3 from Bacillus Megaterium ideal starting materials for the development of designer biocatalysts. Bacterial P450s are used by Sylphium Life Sciences to make all kind of P450 libraries. By applying specific screening assays P450 mutants with the desired bio-catalytic characteristics can be identified and used as a lead enzymes for particular industrial bioconversions.
Construction of a random mutagenic library of a bacterial P450 enzyme was part of an EC supported project. Specially designed expression vectors ware made for this project to obtain high levels of bacterial P450 expression. By using error prone PCR massive P450 mutant libraries were obtained. By using specific screening methods enzymes with particular bio-catalytic characteristics were selected and used for biocatalysis.
