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US 2016/0348133 A1Published application
Method for isomerizing double bonds
AssigneeResearch Foundation of the City University of New York
InventorsEleanore T. Wurtzel, Jesús Beltrán
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Abstract
A method for isomerizing a double bond is provided. A substrate is exposed to an isomerase enzyme, wherein the isomerase enzyme comprises a redox-regulated ligand switch and heme b cofactor. A reducing agent is added which changes iron (III) to iron (II). The enzyme isomerizes double bonds in the iron (II) state but not in the iron (III) state. In one embodiment, the enzyme is homologous with 15-cis-ζ-carotene isomerase (Z-ISO).
Claims17
A method for isomerizing a double bond, the method comprising:
- exposing a substrate to an isomerase enzyme, wherein the isomerase enzyme comprises a redox-regulated ligand switch and heme b cofactor, wherein the heme b cofactor comprises an iron having an iron (III) oxidation state and the substrate comprises a double bond in the substrate with a cis stereochemistry;
- exposing the isomerase enzyme to a reducing agent such that the isomerase enzyme changes from the iron (III) oxidation state to an iron (II) oxidation state, wherein the isomerase enzyme isomerizes a double bond when in the iron (II) oxidation state and does not isomerize the double bond when in the iron (III) oxidation state;
- and
- permitting the double bond in the substrate to undergo isomerization from the cis stereochemistry to a trans stereochemistry, wherein the isomerization is catalyzed by the isomerase enzyme.
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A method for isomerizing a double bond, the method comprising sequential steps of:
- exposing a carotenoid substrate to an isomerase enzyme, wherein the isomerase enzyme comprises a redox-regulated ligand switch and heme b cofactor, wherein the heme b cofactor comprises an iron having an iron (III) oxidation state and the carotenoid substrate comprises a double bond in the carotenoid substrate with a cis stereochemistry, wherein the isomerase enzyme has fewer than four hundred residues and comprises SEQ ID NO. 11;
- exposing the isomerase enzyme to a reducing agent in vitro such that the isomerase enzyme changes from the iron (III) oxidation state to an iron (II) oxidation state, wherein the isomerase enzyme isomerizes a double bond when in the iron (II) oxidation state and does not isomerize the double bond when in the iron (III) oxidation state;
- and
- permitting a double bond in the carotenoid substrate to undergo isomerization from the cis stereochemistry to a trans stereochemistry, wherein the isomerization is catalyzed by the isomerase enzyme.
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Dependencies are inferred from the claim text.