P450-BM3 variants with improved activity
The present invention provides improved P450-BM3 variants with improved activity. In some embodiments, the P450-BM3 variants exhibit improved activity over a wide range of substrates.
1. A recombinant polynucleotide sequence encoding a recombinant cytochrome P450-BM3 variant, wherein said sequence comprises SEQ ID NO: 3, and further wherein said recombinant cytochromie P450-BM3 variant oxidizes at least three organic substrates.
2. An expression vector comprising the polynucleotide sequence of claim 1 .
3. The vector of claim 2 , wherein said polynucleotide sequence is operably linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell.
4. The vector of claim 3 , wherein said host cell is a prokaryotic or eukaryotic cell.
5. The vector of claim 4 , wherein said host cell is a prokaryotic cell.
6. The vector of claim 5 , wherein said host cell is E. coli.
7. A host cell comprising the vector of claim 2 .
8. A method for producing at least one recombinant cytochrome P450-BM3 variant comprising culturing a host cell under conditions such that at least one recombinant cytochrome P450-BM3 variant of claim 1 is produced by said host cell.
9. The method of claim 8 , further comprising the step of recovering said at least one recombinant cytochrome P450 variant.