P450-BM3 variants with improved activity
US-10704030-B2 · Jul 7, 2020 · US
US10982197B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10982197-B2 |
| Application number | US-202016890408-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jun 2, 2020 |
| Priority date | Jul 7, 2015 |
| Publication date | Apr 20, 2021 |
| Grant date | Apr 20, 2021 |
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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.
Opening claim text (preview).
What is claimed is: 1. A recombinant cytochrome P450-BM3 variant comprising the polypeptide sequence set forth in SEQ ID NO: 14. 2. The recombinant cytochrome P450-BM3 variant of claim 1 , wherein said variant oxidizes at least three organic substrates. 3. The recombinant cytochrome P450-BM3 variant of claim 2 , wherein said organic substrate is selected from loratadine, imatinib, geftinib, and diclofenac. 4. A recombinant polynucleotide sequence, wherein said sequence comprises SEQ ID NO: 13. 5. An expression vector comprising the polynucleotide sequence of claim 4 . 6. The vector of claim 5 , wherein said polynucleotide sequence is operably linked with regulatory sequences suitable for expression of said polynucleotide sequence in a suitable host cell. 7. The vector of claim 6 , wherein said host cell is a prokaryotic or eukaryotic cell. 8. The vector of claim 7 , wherein said host cell is a prokaryotic cell. 9. The vector of claim 8 , wherein said host cell is E. coli. 10. A host cell comprising the vector of claim 5 . 11. 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. 12. The method of claim 11 , further comprising the step of recovering said at least one recombinant cytochrome P450-BM3 variant.
Genes encoding for enzymes or proenzymes · CPC title
NADPH-hemoprotein reductase (1.6.2.4), i.e. NADP-cytochrome P450-reductase · CPC title
NADPH-cytochrome P450 reductase (1.6.2.4) · CPC title
Unspecific monooxygenase (1.14.14.1) · CPC title
acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title
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