Reverse beta oxidation pathway
AssigneeWilliam Marsh Rice University
InventorsRamon Gonzalez, James Clomburg, Clementina Dellomonaco, Elliot N. Miller
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Abstract
The invention relates to recombinant microorganisms that have been engineered to produce various chemicals using genes that have been repurposed to create a reverse beta oxidation pathway. Generally speaking, the beta oxidation cycle is expressed and driven in reverse by modifying various regulation points for as many cycles as needed, and then the CoA thioester intermediates are converted to useful products by the action of termination enzymes.
Claims7
An engineered microorganism, wherein the microorganism is engineered to overexpress β-oxidation cycle enzymes and one or more termination enzymes as compared to a corresponding wild-type microorganism, wherein said β-oxidation cycle enzymes comprise:
- a) a thiolase catalyzing the conversion of (Cn)-acyl CoA to β-ketoacyl-CoA,
- b) a hydroxyacyl-CoA dehydrogenase catalyzing the conversion of β-ketoacyl-CoA to β-hydroxyacyl-CoA,
- c) an enoyl-CoA hydratase catalyzing the conversion of β-hydroxyacyl-CoA to trans-Δ2-Enoyl-CoA, and
- d) an acyl-CoA dehydrogenase or transenoyl-CoA reductase catalyzing the conversion of trans-Δ2-enoyl-CoA to (Cn+2)-acyl CoA,
- wherein said β-oxidation cycle runs in the reverse/biosynthetic direction, as recited in enzyme reactions a) through d),
- and
- wherein said one or more termination enzymes are selected from the group consisting of an alcohol-forming coenzyme-A thioester reductase;
- an aldehyde-forming CoA thioester reductase and an alcohol dehydrogenase;
- a thioesterase;
- an acyl-CoA:acetyl-CoA transferase;
- a phosphotransacylase and a carboxylate kinase;
- an aldehyde-forming CoA thioester reductase and an aldehyde decarbonylase;
- and OleA, OleB, OleC, and OleD.
An engineered microorganism comprising a genotype, wherein said genotype comprises fadR, atoC(c), ΔarcA, Δcrp, crp*.
Dependencies are inferred from the claim text.