Regio- and enantioselective alkane hydroxylation with modified cytochrome P450
US7863030B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 15, 2009 |
| Grant date | Jan 4, 2011 |
| Priority date | — |
| Expiry date | Apr 15, 2029 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y114/14001
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Cytochrome P450 BM-3 from Bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant. Mutant 9-10A-A328V hydroxylates octane primarily at the 2-positio to form S-2-octanol (40% ee). Another mutant, 1-12G, hydroxylates alkanes larger than hexane primarily at the 2-position, but forms R-2-alcohols (40-55% ee). These biocatalysts are highly active for alkane substrates and support thousands of product turnovers. These regio- and enantio-selectivities are retained in whole-cell biotransformations with E. coli, where the engineered P450s can be expressed at high levels and the expensive cofactor is supplied endogenously.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.