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US 2020/0048662 A1Published application
Microorganisms and methods for the co-production of ethylene glycol and isobutene
AssigneeBraskem S.A.
InventorsFelipe GALZERANI, Daniel Johannes KOCH, Mateus Schreiner LOPES
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Abstract
The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and isobutene. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and isobutene. Also provided are methods of producing MEG and isobutene using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and isobutene.
Claims277
A recombinant microorganism capable of co-producing monoethylene glycol (MEG) and isobutene from exogenous D-xylose, wherein the recombinant microorganism expresses one or more of the following from (a) to (d):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a D-tagatose 3-epimerase that catalyzes the conversion of D-xylulose to D-ribulose;
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulokinase that catalyzes the conversion of D-ribulose from (a) to D-ribulose-1-phosphate,
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulose-1-phosphate aldolase that catalyzes the conversion of D-ribulose-1-phosphate from (b) to glycolaldehyde and dihydroxyacetonephosphate (DHAP);
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (c) to MEG;
- wherein the recombinant microorganism further expresses one or more of the following from (e) to (h):
- (e) at least one endogenous or exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (e) to acetoacetate;
- (g) at least one endogenous or exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (f) to acetone;
- (h) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovalerate synthase that catalyzes the conversion of acetone from (g) and acetyl-CoA to 3-hydroxyisovalerate (3HIV);
- or
- wherein the recombinant microorganism expresses one or more of the nucleic acid molecule from (a) to (d) above and further expresses one or more of the following from (i) to (n):
- (i) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (i) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (k) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (j) to 3-methylglutaconyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (k) to 3-methylcrotonyl-CoA;
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (l) to 3-hydroxyisovaleryl-CoA;
- (n) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (m) to 3HIV;
- wherein the recombinant microorganism further expresses (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (h) or (n) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3-phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (h) or (n) to isobutene;
- wherein the produced intermediate DHAP is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
19 more elements
A recombinant microorganism capable of co-producing monoethylene glycol (MEG) and isobutene from exogenous D-xylose, wherein the recombinant microorganism expresses one or more of the following from (a) to (c):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a D-xylulose 1-kinase that catalyzes the conversion of D-xylulose to D-xylulose-1-phosphate;
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a D-xylulose-1-phosphate aldolase that catalyzes the conversion of D-xylulose-1-phosphate from (a) to glycolaldehyde and dihydroxyacetonephosphate (DHAP);
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (b) to MEG;
- wherein the recombinant microorganism further expresses one or more of the following from (d) to (g):
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (e) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (d) to acetoacetate;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (e) to acetone;
- (g) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovalerate synthase that catalyzes the conversion of acetone from (f) and acetyl-CoA to 3-hydroxyisovalerate (3HIV);
- or
- wherein the recombinant microorganism expresses one or more of the nucleic acid molecule from (a) to (c) above and further expresses one or more of the following from (h) to (m):
- (h) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (i) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (h) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (i) to 3-methylglutaconyl-CoA;
- (k) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (j) to 3-methylcrotonyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (k) to 3-hydroxyisovaleryl-CoA;
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (l) to 3HIV;
- wherein the recombinant microorganism further expresses (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (g) or (m) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3-phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (g) or (m) to isobutene;
- and wherein the produced intermediate DHAP is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
18 more elements
A recombinant microorganism capable of co-producing monoethylene glycol (MEG) and isobutene from exogenous D-xylose, wherein the recombinant microorganism expresses one or more of the following from (a) to (c):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a xylose dehydrogenase that catalyzes the conversion of D-xylose to D-xylonolactone,
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a xylonolactonase that catalyzes the conversion of D-xylonolactone from (a) to D-xylonate,
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a xylose dehydrogenase that catalyzes the conversion of D-xylose to D-xylonate;
- wherein the recombinant microorganism further expresses one or more of the following from (d) to (f):
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a xylonate dehydratase that catalyzes the conversion of D-xylonate from (b) or (c) to 2-keto-3-deoxy-xylonate;
- (e) at least one endogenous or exogenous nucleic acid molecule encoding a 2-keto-3-deoxy-D-pentonate aldolase that catalyzes the conversion of 2-keto-3-deoxy-xylonate from (d) to glycolaldehyde and pyruvate;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (e) to MEG;
- wherein the recombinant microorganism further expresses one or more of the following from (g) to (j):
- (g) at least one exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (h) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (g) to acetoacetate;
- (i) at least one exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (h) to acetone;
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovalerate synthase that catalyzes the conversion of acetone from (i) and acetyl-CoA to 3-hydroxy-isovalerate (3HIV);
- or
- wherein the recombinant microorganism expresses one or more of the nucleic acid molecule from (a) to (c) above and one or more of the nucleic acid molecule from (d) to (f) above, and further expresses one or more of the following from (k) to (p):
- (k) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (k) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (l) to 3-methylglutaconyl-CoA;
- (n) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (m) to 3-methylcrotonyl-CoA;
- (o) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (n) to 3-hydroxyisovaleryl-CoA;
- (p) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (o) to 3HIV;
- wherein the recombinant microorganism further expresses (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (j) or (p) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3-phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (j) or (p) to isobutene;
- and wherein the produced intermediate pyruvate is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
22 more elements
A method of producing MEG and isobutene using a recombinant microorganism of any of the preceding claims, wherein the method comprises cultivating the recombinant microorganism in a culture medium containing a feedstock providing a carbon source until the MEG and isobutene are produced.
A method of producing a recombinant microorganism that produces or accumulates MEG and isobutene from exogenous D-xylose, comprising introducing into the recombinant microorganism and/or overexpressing one or more of the following from (a) to (d):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a D-tagatose 3-epimerase that catalyzes the conversion of D-xylulose to D-ribulose;
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulokinase that catalyzes the conversion of D-ribulose from (a) to D-ribulose-1-phosphate,
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulose-1-phosphate aldolase that catalyzes the conversion of D-ribulose-1-phosphate from (b) to glycolaldehyde and dihydroxyacetonephosphate (DHAP);
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (c) to MEG;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (e) to (h):
- (e) at least one endogenous or exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (e) to acetoacetate;
- (g) at least one endogenous or exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (f) to acetone;
- (h) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetone from (g) and acetyl-CoA to 3-hydroxyisovalerate (3HIV);
- or
- wherein the method comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the nucleic acid molecule from (a) to (d) above and further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (i) to (n):
- (i) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (i) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (k) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (j) to 3-methylglutaconyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (k) to 3-methylcrotonyl-CoA;
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (l) to 3-hydroxyisovaleryl-CoA,
- (n) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (m) to 3HIV;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (h) or (n) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (h) or (n) to isobutene;
- and wherein the produced intermediate DHAP is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
19 more elements
A method of producing a recombinant microorganism that produces or accumulates MEG and isobutene from exogenous D-xylose, comprising introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (a) to (c):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a D-xylulose 1-kinase that catalyzes the conversion of D-xylulose to D-xylulose-1-phosphate;
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a D-xylulose-1-phosphate aldolase that catalyzes the conversion of D-xylulose-1-phosphate from (a) to glycolaldehyde and dihydroxyacetonephosphate (DHAP);
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (b) to MEG;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (d) to (g):
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (e) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (d) to acetoacetate;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (e) to acetone;
- (g) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovalerate synthase that catalyzes the conversion of acetone from (f) and acetyl-CoA to 3-hydroxyisovalerate (3HIV);
- or
- wherein the method comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the nucleic acid molecule from (a) to (c) above and further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (h) to (m):
- (h) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (i) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (h) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (i) to 3-methylglutaconyl-CoA;
- (k) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (j) to 3-methylcrotonyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (k) to 3-hydroxyisovaleryl-CoA;
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (l) to 3HIV;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (g) or (m) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (g) or (m) to isobutene;
- and wherein the produced intermediate DHAP is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
18 more elements
A method of producing a recombinant microorganism that produces or accumulates MEG and isobutene from exogenous D-xylose, comprising introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (a) to (c):
- (a) at least one endogenous or exogenous nucleic acid molecule encoding a xylose dehydrogenase that catalyzes the conversion of D-xylose to D-xylonolactone;
- and
- (b) at least one endogenous or exogenous nucleic acid molecule encoding a xylonolactonase that catalyzes the conversion of D-xylonolactone from (a) to D-xylonate;
- (c) at least one endogenous or exogenous nucleic acid molecule encoding a xylose dehydrogenase that catalyzes the conversion of D-xylose to D-xylonate;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (d) to (f):
- (d) at least one endogenous or exogenous nucleic acid molecule encoding a xylonate dehydratase that catalyzes the conversion of D-xylonate from (b) or (c) to 2-keto-3-deoxy-xylonate;
- (e) at least one endogenous or exogenous nucleic acid molecule encoding a 2-keto-3-deoxy-D-pentonate aldolase that catalyzes the conversion of 2-keto-3-deoxy-xylonate from (d) to glycolaldehyde and pyruvate;
- (f) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (e) to MEG;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (g) to (j):
- (g) at least one exogenous nucleic acid molecule encoding a thiolase or acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (h) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl-CoA:acetoacetate-CoA transferase or acetate:acetoacetyl-CoA hydrolase that catalyzes the conversion of acetoacetyl-CoA from (g) to acetoacetate;
- (i) at least one exogenous nucleic acid molecule encoding an acetoacetate decarboxylase that catalyzes the conversion of acetoacetate from (h) to acetone;
- (j) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovalerate synthase that catalyzes the conversion of acetone from (i) and acetyl-CoA to 3-hydroxy-isovalerate (3HIV);
- or
- wherein the method comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the nucleic acid molecule from (a) to (c) above and one or more of the nucleic acid molecule from (d) to (f) above, and further comprises introducing into and/or overexpressing in the recombinant microorganism one or more of the following from (k) to (p):
- (k) at least one endogenous or exogenous nucleic acid molecule encoding an acetyl coenzyme A acetyltransferase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;
- (l) at least one endogenous or exogenous nucleic acid molecule encoding a hydroxymethylglutaryl-CoA synthase that catalyzes the conversion of acetoacetyl-CoA from (k) and acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA);
- (m) at least one endogenous or exogenous nucleic acid molecule encoding a methylglutaconyl-CoA hydratase that catalyzes the conversion of HMG-CoA from (l) to 3-methylglutaconyl-CoA;
- (n) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA carboxylase that catalyzes the conversion of 3-methylglutaconyl-CoA from (m) to 3-methylcrotonyl-CoA;
- (o) at least one endogenous or exogenous nucleic acid molecule encoding a methylcrotonyl-CoA hydratase that catalyzes the conversion of 3-methylcrotonyl-CoA from (n) to 3-hydroxyisovaleryl-CoA;
- (p) at least one endogenous or exogenous nucleic acid molecule encoding a 3-hydroxyisovaleryl-CoA thioesterase that catalyzes the conversion of 3-hydroxyisovaleryl-CoA from (o) to 3HIV;
- wherein the method further comprises introducing into and/or overexpressing in the recombinant microorganism (a1) and (a2), and/or (b1) selected from:
- (a1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV kinase that catalyzes the conversion of 3HIV from (j) or (p) to 3HIV-3-phosphate;
- (a2) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV-3-phosphate decarboxylase that catalyzes the conversion of 3HIV-3phosphate from (a1) to isobutene;
- (b1) at least one endogenous or exogenous nucleic acid molecule encoding a 3HIV decarboxylase that catalyzes the conversion of 3HIV from (j) or (p) to isobutene;
- and wherein the produced intermediate pyruvate is converted to acetyl-CoA through the endogenous glycolysis pathway in the microorganism, and wherein MEG and isobutene are co-produced.
23 more elements
A recombinant microorganism co-producing monoethylene glycol (MEG) and isobutene.
Dependencies are inferred from the claim text.