Method for enantioselective carbene C—H insertion using an iron-containing protein catalyst
AssigneeCalifornia Institute of Technology
InventorsRuijie Zhang, Lena Wohlschlager, Hans H. Renata, Frances H. Arnold, Kai Chen
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Abstract
Methods for catalyzing C—H insertion reactions using heme enzymes are described. The present disclosure provides a method for producing a C—H insertion product comprising providing an substrate having an sp3-hybridized C—H bond, a carbene precursor such as a diazo reagent, and a heme enzyme, and admixing the components in a reaction for a time sufficient to produce the C—H insertion product. Heme enzyme variants useful for carrying out in vivo and in vitro C—H insertion reactions, as well as expression vectors and host cells expressing the heme enzymes, are also described.
Claims16
A reaction mixture for producing a C—H insertion product, the reaction mixture comprising a substrate having an sp3-hybridized C—H bond, a carbene precursor for modification of the carbon atom in the sp3-hybridized C—H bond, and a heme protein comprising an iron porphyrin,
- wherein the heme protein is selected from the group consisting of a truncated P450BM3 comprising the amino acid sequence set forth in SEQ ID NO:1, or a variant thereof, a full-length P450BM3 comprising the amino acid sequence set forth in SEQ ID NO:2, or a variant thereof, and a nitric oxide dioxygenase protein from Rhodothermus marinus or a variant thereof, and
- wherein the carbene precursor and the substrate having the spa-hybridized C—H bond are separate compounds, or the carbene precursor is present in the same compound as the substrate having the spa-hybridized C—H bond.
A method for producing a C—H insertion product, the method comprising:
- (a) providing a substrate having an sp3-hybridized C—H bond, a carbene precursor for modification of the carbon atom in the sp3-hybridized C—H bond, and a heme protein comprising an iron porphyrin,
- wherein the heme protein is selected from the group consisting of a truncated P450BM3 comprising the amino acid sequence set forth in SEQ ID NO:1, or a variant thereof, a full-length P450BM3 comprising the amino acid sequence set forth in SEQ ID NO:2, or a variant thereof, and a nitric oxide dioxygenase protein from Rhodothermus marinus or a variant thereof;
- and
- (b) admixing the components of step (a) under conditions sufficient to produce the C—H insertion product.
Dependencies are inferred from the claim text.