US6080555A
Expired / Lapsed
US
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Synthesis of polyketides from diketides
Assignee: Brown University Research Foundation Inc
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Filing: 1996-07-05
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Granted: 2000-06-27
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Expires: 2016-07-05
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
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 03:35
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: 15
Claim 1
Independent Claim
111 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; (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
111 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; (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
21 words
The method of claim 1 wherein one amino acid in the catalytic domain of said first module ketosynthase has been altered.
Claim 4
Dependent Claim
11 words
The method of claim 2 wherein said amino acid is cysteine.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein said cysteine is replaced by alanine.
Claim 6
Dependent Claim
14 words
The method of claim 1 wherein said modified PKS is contained in host cells.
Claim 7
Dependent Claim
14 words
The method of claim 5 wherein said cells are heterologous to said modified PKS.
Claim 8
Dependent Claim
10 words
The method of claim 5 wherein said cells are Streptomyces.
Claim 9
Dependent Claim
13 words
The method of claim 5 wherein said cells are permeable to said diketide.
Claim 10
Dependent Claim
20 words
The method of claim 5 wherein said cells have been modified to delete a native PKS contained in said cells.
Claim 11
Dependent Claim
12 words
The method of claim 9 wherein said cells are S. coelicolor CH999.
Claim 12
Dependent Claim
55 words
The method of claim 5 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 13
Dependent Claim
18 words
The method of claim 1 wherein said diketide is in the form of an N-acetyl cysteamine (NAc) thioester.
Claim 14
Dependent Claim
11 words
The method of claim 12 wherein said diketide is (2S,3R)-2-methyl-3-hydroxypentanoyl-NAc thioester.
Claim 15
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.
References Cited (Backward Citations)
US5824513A
Cited
WO1993013663A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-06-27 | 2003-12-27 | 2004-06-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-06-27 | 2007-12-27 | 2008-06-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-06-27 | 2011-12-27 | 2012-06-27 | $7700.00 | Window Closed / Lapsed |