Abstract: |
This disclosure generally relates to the use of microorganisms to make various functionalized polyketides through polyketoacyl-CoA thiolase-catalyzed non-decarboxylative condensation reactions instead of decarboxylative reactions catalyzed by polyketide synthases. Native or engineered polyketoacyl-CoA thiolases catalyze the non-decarboxylative Claisen condensation in an iterative manner (i.e. multiple rounds) between two either unsubstituted or functionalized ketoacyl-CoAs (and polyketoacyl-CoAs) serving as the primers and acyl-CoAs serving as the extender unit to generate (and elongate) polyketoacyl-CoAs. Before the next round of polyketoacyl-CoA thiolase reaction, the β-keto group of the polyketide chain of polyketoacyl-CoA can be reduced and modified step-wise by 3-OH-polyketoacyl-CoA dehydrogenase or polyketoenoyl-CoA hydratase or polyketoenoyl-CoA reductase. Dehydrogenase converts the β-keto group to β-hydroxy group. Hydratase converts the β-hydroxy group to α-β-double-bond. Reductase converts the α-β-double-bond to single bond. Spontaneous or thioesterase catalyzed termination reaction terminates the elongation of polyketide chain of polyketoacyl-CoA at any point through CoA removal and spontaneous reactions rearrange the structure, generating the final functional polyketide products. |
Inventor: |
GONZALEZ, Ramon (Tampa, FL, US); CHEONG, Seokjung (Houston, TX, US); CLOMBURG, James M. (Houston, TX, US) |
Applicant: |
Gonzalez, Ramon (Tampa, FL, US) |
Face Assignee: |
N/A |
Filed: |
2020-07-15 |
Issued: |
2021-02-25 |
Claims: |
20 |
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US20210054427
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1) A method of making a polyketide, comprising growing a genetically engineered microorganism in a nutrient broth for a time sufficient to produce a polyketide and isolating said polyketide or a spontaneously rearranged form of said polyketide or a derivative of said polyketide, wherein said microorganism has a polyketide-producing pathway comprising the following substrate(s) to product(s) conversions:
(3)
(8)
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10) A method, comprising:
(2)
(3)
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13) A genetically engineered microorganism, wherein said microorganism has a polyketide-producing pathway comprising the following substrate(s) to product(s) conversions:
(2)
(7)
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16) A recombinant microorganism having expressed polyketoacyl-CoA thiolase(s) that catalyze(s) the following reaction:
(1)
(1)
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20) A recombinant microorganism having expressed genes encoding at least two different thiolases, said thiolases including a ketoacyl thiolase and a polyketoacyl thiolase. (0)
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