Reverse beta oxidation pathway

1. 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 (C n )-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 (C n+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.

2. The method of claim 1 , further comprising supplementing said nutrient broth with propionate in order to produce odd-chain length products.

3. The method of claim 1 , further comprising growing said engineered microorganism under microaerobic (<10% O 2 ) or anaerobic conditions at a temperature of 30-40° C., wherein said nutrient broth comprises 0-100 μM FeSO 4 and 0-5 mM calcium pantothenate.

4. The method of claim 1 , wherein said one or more olefin-forming enzymes are selected from:

OleA, OleB, OleC, and OleD.

5. The method of claim 4 , wherein said engineered microorganism comprises a genotype selected from the group consisting of:

a) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, ΔydiO, [yqeF+, tesB+];

b) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [fadBA+, fadM+]; and

c) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [fadBA+, yciA+].

6. The method of claim 4 , wherein said engineered microorganism comprises a genotype selected from the group consisting of:

a) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [yqeF+, acrM+, PCC7942_orf1593+];

b) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [yqeF+, acrM+, PCC7942_orf1593+];

c) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [yqeF+, oleABCD+,];

d) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [fadBA+, acrM+, PCC7942_orf1593+];

e) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [fadBA+, acr1+, PCC7942_orf1593+]; and

f) fadR, atoC(c), ΔarcA, Δcrp, crp*, ΔadhE, ΔfrdA, Δpta, ΔyqhA, ΔfucO, ΔfadD, [fadBA+, oleABCD+,].