Thermostabilization of a cytochrome P450 peroxygenase

Abstract

Some like it hot: We previously described a laboratory-evolved variant of the cytochrome P450 BM-3 heme domain which functions as a peroxide-driven hydroxylase (peroxygenase, see picture). This biocatalyst does not require additional proteins or NADPH to drive hydroxylation and is amenable to further improvements through protein engineering. Here we describe a thermostable variant whose half-life at 57.5 °C is 50 times that of the F87A variant and 250 times that of the wildtype holoenzyme.