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AssigneeCalifornia Institute of Technology
InventorsFrances H. Arnold, Matthew W. Peters, Peter Meinhold
Estimating the term…
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.
An isolated mutant cytochrome P450 Bacillus megaterium 3 (BM-3) enzyme comprising the amino acid sequence of SEQ ID NO:2, except that a leucine is at position 82.
An isolated mutant P450 enzyme comprising at least 90% identity to SEQ ID NO:2 and having a A82L point mutation and can hydroxylate an alkane.
An isolated mutant P450 enzyme having at least 90% sequence identity to the P450 enzyme of SEQ ID NO:2, wherein said enzyme comprises a leucine at position 82 of SEQ ID NO:
Dependencies are inferred from the claim text.