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US 2018/0355394 A1Published application
Bioconversion of short-chain hydrocarbons to fuels and chemicals
OwnerRamon Gonzalesinventor
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OwnerRamon Gonzalesinventor
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An engineered microorganism(s) with novel pathways for the conversion of short-chain hydrocarbons to fuels and chemicals (e.g. carboxylic acids, alcohols, hydrocarbons, and their alpha-, beta-, and omega-functionalized derivatives) is described. Key to this approach is the use of hydrocarbon activation enzymes able to overcome the high stability and low reactivity of hydrocarbon compounds through the cleavage of an inert C—H bond. Oxygen-dependent or oxygen-independent activation enzymes can be exploited for this purpose, which when combined with appropriate pathways for the conversion of activated hydrocarbons to key metabolic intermediates, enables the generation of product precursors that can subsequently be converted to desired compounds through established pathways. These novel engineered microorganism(s) provide a route for the production of fuels and chemicals from short chain hydrocarbons such as methane, ethane, propane, butane, and pentane.
A genetically engineered microorganism with one or more expression vectors or integrated sequences encoding overexpressed 1) alkane monooxygenase or alkane hydroxylase (EC 1.14.15.3), 2) alcohol dehydrogenase (EC 1.1.1.-), 3) aldehyde dehydrogenase (EC 1.2.1.-), and 4) acyl-CoA synthetase (EC 6.2.1.-);
A genetically engineered microorganism with one or more expression vectors or integrated sequences encoding overexpressed 1) alkyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-alkyl-malonyl-CoA mutase (EC 5.4.99.-), 4) 2-methyl-alkyl-malonyl-CoA decarboxylase (EC 4.1.1.-), 5) propionyl-CoA carboxylase (EC 6.4.1.3), 6) methylmalonyl-CoA epimerase (EC 5.1.99.1), 7) methylmalonyl-CoA mutase (EC 5.1.99.2), 8) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or succinyl-CoA synthetase (EC 6.2.1.5), and 9) succinate dehydrogenase (EC 1.3.5.1), wherein said microorganism is able to convert a short chain alkane (C1-C5) to an acyl-CoA through fumarate addition to said short chain alkane and subsequent regeneration of said fumarate.
A genetically engineered microorganism with one or more expression vectors or integrated sequences encoding overexpressed 1) methyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-succinyl-CoA dehydrogenase (EC 1.3.99.-), 4) mesaconyl-C1-CoA-C4-CoA transferase (EC 2.8.3.-), 5) mesaconyl-C4-CoA hydratase (EC 4.2.1.153), 6) L-malyl-CoA/citramalyl-CoA lyase (EC 4.1.3.25), 7) pyruvic-malic carboxylase (EC 1.1.1.39), and 8) fumarase (EC 4.2.1.2), wherein said microorganism is able to convert methane to an acyl-coA through fumarate addition to said methane and subsequent regeneration of said fumarate.
A genetically engineered microorganism with one or more expression vectors encoding overexpressed 1) methyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-alkyl-succinyl-CoA dehydrogenase (EC 1.3.99.-), 4) mesaconyl-CoA hydratase/β-methylmalyl-CoA dehydratase (EC 4.2.1.148), 5) β-methylmalyl-CoA lyase (EC 4.1 3.24), 6) propionyl-CoA carboxylase (EC 6.4.1.3), 7) methylmalonyl-CoA epimerase (EC 5.1.99.1) and methylmalonyl-CoA mutase (EC 5.1.99.2), 8) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or succinyl-CoA synthetase (EC 6.2.1.5), 9) succinate dehydrogenase (EC 1.3.5.1), 10:
A genetically engineered microorganism being E. coli and comprising one or more expression vectors or integrated sequences encoding overexpressed 1) alkane monooxygenase or alkane hydroxylase (EC 1.14.15.3), 2) alcohol dehydrogenase (EC 1.1.1.-), 3) aldehyde dehydrogenase (EC 1.2.1.-), and 4) acyl-CoA synthetase (EC 6.2.1.-);
A genetically engineered microorganism being E. coli and comprising one or more expression vectors or integrated sequences encoding overexpressed 1) alkyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-alkyl-malonyl-CoA mutase (EC 5.4.99.-), 4) 2-methyl-alkyl-malonyl-CoA decarboxylase (EC 4.1.1.-), 5) propionyl-CoA carboxylase (EC 6.4.1.3), 6) methylmalonyl-CoA epimerase (EC 5.1.99.1), 7) methylmalonyl-CoA mutase (EC 5.1.99.2), 8) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or succinyl-CoA synthetase (EC 6.2.1.5), and 9) succinate dehydrogenase (EC 1.3.5.1), wherein said microorganism is able to converts a short chain alkane (C1-C5) to an acyl-CoA through fumarate addition to said short chain alkane and subsequent regeneration of said fumarate.
A genetically engineered microorganism being E. coli and comprising one or more expression vectors or integrated sequences encoding overexpressed 1) methyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-succinyl-CoA dehydrogenase (EC 1.3.99.-), 4) mesaconyl-C1-CoA-C4-CoA transferase (EC 2.8.3.), 5) mesaconyl-C4-CoA hydratase (EC 4.2.1.153), 6) L-malyl-CoA/citramalyl-CoA lyase (EC 4.1.3.25), 7) pyruvic-malic carboxylase (EC 1.1.1.39), and 8) fumarase (EC 4.2.1.2), wherein said microorganism is able to convert methane to an acyl-coA through fumarate addition to said methane and subsequent regeneration of said fumarate.
A genetically engineered microorganism being E. coli and comprising one or more expression vectors or integrated sequences encoding overexpressed 1) methyl succinate synthase (EC 4.1.-), 2) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or 2-methyl-alkyl-succinyl-CoA synthetase (EC 6.2.1.-), 3) 2-methyl-alkyl-succinyl-CoA dehydrogenase (EC 1.3.99.-), 4) mesaconyl-CoA hydratase/β-methylmalyl-CoA dehydratase (EC 4.2.1.148), 5) β-methylmalyl-CoA lyase (EC 4.1.3.24), 6) propionyl-CoA carboxylase (EC 6.4.1.3), 7) methylmalonyl-CoA epimerase (EC 5.1.99.1) and methylmalonyl-CoA mutase (EC 5.1.99.2), 8) succinyl-CoA:2-methyl-alkyl-succinyl-CoA transferase (EC 2.8.3.-) or succinyl-CoA synthetase (EC 6.2.1.5), 9) succinate dehydrogenase (EC 1.3.5.1), 10:
A genetically engineered microorganism converting a short-chain (C1-C5) alkane substrate to a product, said microorganism comprising enzymes or overexpressed enzymes catalyzing:
A genetically engineered microorganism converting a short-chain (C1-C5) alkane substrate to a product, said microorganism comprising enzymes or overexpressed enzymes catalyzing:
Dependencies are inferred from the claim text.