Reverse beta oxidation pathway

A method of making a product, comprising growing an engineered microorganism in a nutrient broth for a time sufficient to make a product, and isolating said product, wherein said product is selected from the group consisting of carboxylic acids, alkanes, or alkenes, wherein said engineered microorganism comprises:

• a) overexpression of β-oxidation cycle enzymes as compared to a corresponding wild type microorganism, wherein said enzymes comprise:
• i) a thiolase catalyzing the conversion of (Cn)-acyl CoA to β-ketoacyl-CoA;
• ii) a hydroxyacyl-coA dehydrogenase catalyzing the conversion of β-ketoacyl-CoA to β-hydroxyacyl-CoA;
• iii) an enoyl-coA hydratase catalyzing the conversion of β-hydroxyacyl-CoA to trans-Δ2-enoyl-coA;
• and
• iv) an acyl-CoA dehydrogenase or a transenoyl-CoA reductase catalyzing the conversion of trans-Δ2-enoyl-coA to (Cn+2)-acyl CoA;
• b) functional operation of a β-oxidation cycle in a reverse biosynthetic direction as recited in steps i) to iv);
• and
• c) overexpression of one or more termination enzyme(s) as compared to a corresponding wild type microorganism, wherein said termination enzyme(s) are selected from:
• i) an alcohol-forming coenzyme-A thioester reductase, or an aldehyde-forming CoA thioester reductase plus an alcohol dehydrogenase, to convert intermediates produced by reversal of the β-oxidation cycle to trans Δ2 fatty alcohols, β-keto alcohols, 1,3 diols, or β-hydroxy acids;
• ii) a thioesterase, or an acyl-CoA:acetyl-CoA transferase, or a phosphotransacylase and a carboxylate kinase, to convert intermediates produced by reversal of the β-oxidation cycle to carboxylic acids;
• iii) an aldehyde-forming CoA thioester reductase and an aldehyde decarbonylase, to convert intermediates produced by reversal of the β-oxidation cycle to alkanes or terminal alkenes;
• and
• iv) one or more olefin-forming enzymes to convert intermediates produced by reversal of the β-oxidation cycle to alkenes.