Cytosolic isobutanol pathway localization for the production of isobutanol
DC
First Claim
1. A recombinant yeast microorganism for producing isobutanol, the recombinant yeast microorganism comprising an isobutanol producing metabolic pathway, wherein said isobutanol producing metabolic pathway comprises the following substrate to product conversions:
- (i) pyruvate to acetolactate;
(ii) acetolactate to 2,3-dihydroxyisovalerate;
(iii) 2,3-dihydroxyisovalerate to α
-ketoisovalerate;
(iv) α
-ketoisovalerate to isobutyraldehyde; and
(v) isobutyraldehyde to isobutanol;
wherein said recombinant yeast microorganism expresses;
an exogenous nucleic acid encoding dihydroxy acid dehydratase to catalyze the conversion of 2,3-dihydroxyisovalerate to α
-ketoisovalerate, said exogenous nucleic acid encoding dihydroxy acid dehydratase comprising an amino acid motif which is at least 90% identical to the motif of SEQ ID NO;
27, wherein said dihydroxy acid dehydratase is active in the cytosol of said recombinant yeast microorganism, and wherein the recombinant yeast microorganism has been engineered to inactivate one or more endogenous pyruvate decarboxylase (PDC) genes.
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Abstract
The present invention provides recombinant microorganisms comprising isobutanol producing metabolic pathway with at least one isobutanol pathway enzyme localized in the cytosol, wherein said recombinant microorganism is selected to produce isobutanol from a carbon source. Methods of using said recombinant microorganisms to produce isobutanol are also provided. In various aspects of the invention, the recombinant microorganisms may comprise a cytosolically active isobutanol pathway enzymes. In some embodiments, the invention provides mutated, modified, and/or chimeric isobutanol pathway enzymes with cytosolic activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.
65 Citations
20 Claims
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1. A recombinant yeast microorganism for producing isobutanol, the recombinant yeast microorganism comprising an isobutanol producing metabolic pathway, wherein said isobutanol producing metabolic pathway comprises the following substrate to product conversions:
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(i) pyruvate to acetolactate; (ii) acetolactate to 2,3-dihydroxyisovalerate; (iii) 2,3-dihydroxyisovalerate to α
-ketoisovalerate;(iv) α
-ketoisovalerate to isobutyraldehyde; and(v) isobutyraldehyde to isobutanol; wherein said recombinant yeast microorganism expresses; an exogenous nucleic acid encoding dihydroxy acid dehydratase to catalyze the conversion of 2,3-dihydroxyisovalerate to α
-ketoisovalerate, said exogenous nucleic acid encoding dihydroxy acid dehydratase comprising an amino acid motif which is at least 90% identical to the motif of SEQ ID NO;
27, wherein said dihydroxy acid dehydratase is active in the cytosol of said recombinant yeast microorganism, and wherein the recombinant yeast microorganism has been engineered to inactivate one or more endogenous pyruvate decarboxylase (PDC) genes.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification
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- Resources
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Current AssigneeGevo Incorporated
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Original AssigneeGevo Incorporated
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InventorsUrano, Jun, Dundon, Catherine Asleson
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Primary Examiner(s)Saidha, Tekchand
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Assistant Examiner(s)Meah, MD. Younus
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Application NumberUS12/855,276Publication NumberTime in Patent Office719 DaysField of SearchNoneUS Class Current435/254.2CPC Class CodesC12N 15/81 for yeastsC12N 9/0006 acting on CH-OH groups as d...C12N 9/1022 transferring aldehyde or ke...C12N 9/88 Lyases (4.)C12P 7/16 ButanolsY02E 50/10 Biofuels, e.g. bio-diesel
