Structural analysis of Cytochrome P450 BM3 mutant M11 in complex with dithiothreitol

Role of Residue 87 in Substrate- and Regioselectivity of Drug Metabolizing Cytochrome P450 Bm3 M11

BM3, originating from Bacillus megaterium, is a highly active enzyme which has attracted much attention because of its potential applicability as a biocatalyst for oxidative reactions. Previously we developed a drug metabolizing mutant BM3 M11 by a combination of site-directed and random mutagenesis. BM3 M11 contains ten mutations, when compared to wild-type BM3 and is able to produce human-rel...

On-line diversity screening of cytochrome P450 BM3 mutant libraries

Photooxidation of cytochrome P450-BM3.

High-valent iron-oxo species are thought to be intermediates in the catalytic cycles of oxygenases and peroxidases. An attractive route to these iron-oxo intermediates involves laser flash-quench oxidation of ferric hemes, as demonstrated by our work on the ferryl (compound II) and ferryl porphyrin radical cation (compound I) intermediates of horseradish peroxidase. Extension of this work to in...

Protein-surfactant film voltammetry of wild-type and mutant cytochrome P450 BM3.

We are investigating the redox chemistry of wild-type (WT) and mutant (1-12G) cytochrome P450 BM3. Absorption spectra in solution feature the Fe(III) Soret at 418 nm for WT and a split Soret for 1-12G at 390 and 418 nm. Voltammetry of the proteins within DDAPSS films on the surface of carbon electrodes reveal nearly identical Fe(III/II) potentials (approximately -200 mV vs Ag/AgCl), but signifi...

Probing Steroidal Substrate Specificity of Cytochrome P450 BM3 Variants.

M01A82W, M11A82W and M01A82WS72I are three cytochrome P450 BM3 (CYP102A1) variants. They can catalyze the hydroxylation of testosterone (TES) and norethisterone at different positions, thereby making them promising biocatalysts for steroid hydroxylation. With the aim of obtaining more hydroxylated steroid precursors it is necessary to probe the steroidal substrate diversity of these BM3 variant...