Patent US6274560B1 successfully refreshed from Google Patents.
US6274560B1
Expired / Lapsed
US
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Cell-free synthesis of polyketides
Assignee: Brown University Research Foundation Inc
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Filing: 1999-08-13
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Granted: 2001-08-14
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Expires: 2019-08-13
Abstract
Abstract Cell-free systems which effect the production of polyketides employing modular polyketide synthases are described. Libraries of new and/or known polyketides may also be produced in cell-free systems employing aromatic PKS, modular PKS or both.
Inventors
- 👤 Chaitan Khosla
- 👤 Rembert Pieper
- 👤 Guanglin Luo
- 👤 David E. Cane
- 👤 Camilla Kao
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 23:53
CPC Classifications
C; C07; C07H; C07H17/00; C07H17/04; C07H17/08; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/52; C12P; C12P17/00; C12P17/02; C12P17/06; C12P17/08
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
126 words
A method to produce a desired polyketide which method comprises: (a) providing a functional modular polyketide synthase (PKS) comprising at least a first and second module wherein said PKS has been modified to prevent its utilization of a native starter unit for said modular PKS by inactivation of the catalytic domain of the ketosynthase of said first module, but wherein said PKS is able to incorporate a diketide substrate into at least a triketide, and which PKS is a complete erythromycin PKS gene cluster, i.e., 6-deoxyerythronolide B (6-dEB) synthase; (b) adding to said modified PKS a diketide that is a substrate for the modified PKS; (c) incubating the modified PKS and said diketide under conditions wherein said polyketide is synthesized; and (d) optionally recovering the polyketide.
Claim 2
Independent Claim
126 words
A method to produce a desired polyketide which method comprises: (a) providing a functional modular polyketide synthase (PKS) comprising at least a first and second module wherein said PKS has been modified to prevent its utilization of a native starter unit for said modular PKS by inactivation of the catalytic domain of the ketosynthase of said first module, but wherein said PKS is able to incorporate a diketide substrate into at least a triketide, and which PKS is a complete erythromycin PKS gene cluster, i.e., 6-deoxyerythronolide B (6-dEB) synthase; (b) adding to said modified PKS a diketide that is a substrate for the modified PKS; (c) incubating the modified PKS and said diketide under conditions wherein said polyketide is synthesized; and (d) optionally recovering the polyketide.
Claim 3
Dependent Claim
14 words
The method of claim 1 wherein said modified PKS is contained in host cells.
Claim 4
Dependent Claim
45 words
The method of claim 2 wherein said cells are heterologous to said modified PKS, and/or wherein said cells are Streptomyces; and/or wherein said cells are permeable to said diketide; and/or wherein said cells have been modified to delete a native PKS contained in said cells.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein said cells are S. coelicolor CH999.
Claim 6
Dependent Claim
13 words
The method of claim 1 which is conducted in a cell free system.
Claim 7
Dependent Claim
55 words
The method of claim 1 wherein the diketide is that obtained by the coupling of a starter unit which is acetyl CoA, malonamyl Co-A, propionyl Co-A, butyryl Co-A, isobutyryl Co-A, isovaleryl Co-A, benzoyl Co-A, aminobenzoyl Co-A, aminohydroxybenzoyl Co-A, or thiophene carboxyl Co-A, with an extender unit which is malonyl Co-A, methylmalonyl Co-A or ethylmalonyl Co-A.
Claim 8
Dependent Claim
46 words
The method of claim 6 wherein the diketide is that obtained by the coupling of a starter unit which is malonamyl Co-A, butyryl Co-A, isobutyryl Co-A, isovaleryl Co-A, benzoyl Co-A, aminobenzoyl Co-A, or aminohydroxybenzoyl Co-A with an extender unit which is methylmalonyl Co-A or ethylmalonyl Co-A.
Claim 9
Dependent Claim
37 words
The method of claim 7 wherein the diketide is that obtained by the coupling of a starter unit which is malonamyl Co-A, butyryl Co-A, isobutyryl Co-A or isovaleryl Co-A with an extender unit which is methylmalonyl Co-A.
Claim 10
Dependent Claim
30 words
The method of claim 8 wherein the diketide is that obtained by the coupling of a starter unit which is butyryl Co-A with an extender unit which is methylmalonyl Co-A.
Claim 11
Dependent Claim
10 words
A polyketide prepared by the method of claim 7 .
Claim 12
Dependent Claim
10 words
A polyketide prepared by the method of claim 8 .
Claim 13
Dependent Claim
10 words
A polyketide prepared by the method of claim 9 .
Claim 14
Dependent Claim
8 words
The polyketide of claim 12 which is 13-propyl-6-dEB.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2004-08-13 | 2005-02-12 | 2005-08-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-08-13 | 2009-02-12 | 2009-08-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-08-13 | 2013-02-12 | 2013-08-14 | $7700.00 | Window Closed / Lapsed |