Microorganisms and methods for the biosynthesis of (2-hydroxy-3methyl-4-oxobutoxy) phosphonate

A non-naturally occurring Escherichia coli comprising:

• a reductive tricarboxylic acid (rTCA) pathway comprising at least one exogenous nucleic acid encoding a rTCA pathway enzyme expressed in a sufficient amount to convert (i) CO, (ii) CO2 and H2, (iii) CO and CO2, (iv) synthesis gas comprising CO and H2, or (v) synthesis gas comprising CO, CO2, and H2 to acetyl-CoA, wherein the rTCA pathway enzyme is selected from the group consisting of an ATP citrate-lyase, a citrate lyase, an aconitase, an isocitrate dehydrogenase, an alpha-ketoglutarate:ferredoxin oxidoreductase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarate reductase, a fumarase and a malate dehydrogenase;
• a reducing equivalent producing pathway comprising at least one exogenous nucleic acid encoding a reducing equivalent producing enzyme selected from the group consisting of a carbon monoxide dehydrogenase, a hydrogenase and a NAD(P)H:ferredoxin oxidoreductase;
• at least one exogenous nucleic acid encoding a (2-hydroxy-3-methyl-4 oxobutoxy)phosphonate (2H3M40P) pathway enzyme expressed in a sufficient amount to produce (2H3M40P), wherein the (2H3M40P) pathway enzyme is selected from the group consisting of 1-deoxy-D-xylulose-5-phosphate (DXP) synthase, DXP reductoisomerase;
• and 2ME4P dehydratase;
• at least one exogenous nucleic acid encoding a p-toluate pathway enzyme in a sufficient amount to produce p-toluate, wherein the p-toluate pathway enzyme is selected from the group consisting of 2-dehydro-3-deoxyphosphoheptonate synthase;
• 3-dehydroquinate synthase;
• 3-dehydroquinate dehydratase;
• shikimate dehydrogenase;
• shikimate kinase;
• 3-phosphoshikimate-2-carboxyvinyltransferase;
• chorismate synthase;
• and chorismate lyase;
• and
• at least one exogenous nucleic acid encoding a terephthalic acid (PTA) pathway enzyme expressed in a sufficient amount to produce PTA wherein the PTA pathway enzyme is selected from the group consisting of p-toluate methyl-monooxygenase reductase, 4-carboxybenzl alcohol dehydrogenase and 4-carboxybenzyl aldehyde dehydrogenase;
• and
• wherein the Escherichia coli does not require acetyl-CoA synthase;
• and wherein the biosynthetic product is p-toluate, terephthalate or (2-hydroxy-3-methyl-4 oxobutoxy)phosphonate.

A non-naturally occurring Escherichia coli comprising:

• a reducing equivalent producing pathway comprising at least one exogenous nucleic acid encoding a reducing equivalent producing enzyme selected from the group consisting of a carbon monoxide dehydrogenase, a hydrogenase and a NAD(P)H:ferredoxin oxidoreductase, and at least one exogenous nucleic acid encoding a biosynthetic product pathway enzyme expressed in a sufficient amount to produce the biosynthetic product, wherein the Escherichia coli does not require acetyl-CoA synthase;
• and wherein biosynthetic product is p-toluate, terephthalate (PTA) or (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate (2H3M40P);
• wherein the 2H3M40P pathway enzyme is expressed in a sufficient amount to produce 2H3M40P, wherein the (2H3M40P) pathway enzyme is selected from the group consisting of 1-deoxy-D-xylulose-5-phosphate (DXP) synthase, DXP reductoisomerase, and 2ME4P dehydratase;
• wherein the p-toluate pathway enzyme is expressed in a sufficient amount to produce p-toluate, wherein the p-toluate pathway enzyme is selected from the group consisting of 2-dehydro-3-deoxyphosphoheptonate synthase;
• 3-dehydroquinate synthase;
• 3-dehydroquinate dehydratase;
• shikimate dehydrogenase;
• shikimate kinase;
• 3-phosphoshikimate-2-carboxyvinyltransferase;
• chorismate synthase;
• and chorismate lyase;
• and
• wherein the PTA pathway enzyme expressed in a sufficient amount to produce PTA wherein the PTA pathway enzyme is selected from the group consisting of p-toluate methyl-monooxygenase reductase, 4-carboxybenzl alcohol dehydrogenase and 4-carboxybenzyl aldehyde dehydrogenase.