TYPE II FATTY ACID SYNTHESIS ENZYMES IN REVERSE ß-OXIDATION

摘要

This disclosure describes enzymes from the type II (a discrete set of enzymes) fatty acid synthesis (“FAS”) pathway that can be used in combination with thiolases to operate a functional reversal of the β-oxidation cycle. A combination of thiolases with one or more of 3-oxoacyl-[acyl-carrier-protein] reductase (FabG, others), 3-hydroxyacyl-[acp] dehydratase (FabA, FabZ, others), and enoyl-[acyl-carrier-protein] reductase (FabI, FabK, FabL, FabV, others) yields a functional reversal of the β-oxidation cycle. If only one or two enzymes are used, the remaining enzymes will be traditional beta oxidation enzymes. Once this cycle is coupled with the appropriate priming and termination pathways, the production of carboxylic acids, alcohols, hydrocarbons, amines and their α-, β-, and ω-functionalized derivatives from renewable carbon sources can be achieved.

主权项

1. A genetically engineered microorganism, said microorganism having a reverse beta oxidation (BOX-R) cycle that grows a primer by adding a 2-carbon donar thereto in each cycle, said BOX-R cycle comprising:
a. an overexpressed thiolase that catalyzes the non-decarboxylative condensation of an acyl-CoA primer with a 2-carbon donor acetyl-CoA to produce a β-ketoacyl-CoA; b. an overexpressed 3-oxoacyl-[acyl-carrier-protein] reductase or overexpressed 3-hydroxyacyl-CoA dehydrogenase that catalyzes the reduction of a β-ketoacyl-CoA to a β-hydroxyacyl-CoA; c. an overexpressed 3-hydroxyacyl-[acyl-carrier-protein] dehydratase or an overexpressed enoyl-CoA hydratase or 3-hydroxyacyl-CoA dehydratase that catalyzes the dehydration of a (3R)-β-hydroxyacyl-CoA to a transenoyl-CoA; d. an overexpressed enoyl-[acyl-carrier-protein] reductase or acyl-CoA dehydrogenase or trans-enoyl-CoA reductase that catalyzes the reduction of a transenoyl-CoA to an acyl-CoA that is two carbons longer than said acyl coA primer;
i. wherein at least one of the enzymes from steps b-d is a fatty acid synthesis enzyme; e. an overexpressed termination pathway that catalyzed the exit of an intermediate from said BOX-R cycle.