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US 6,274,560 B1Granted
Cell-free synthesis of polyketides
AssigneesBrown University Research Foundation, The Board of Trustees of the Leland Stanford Junior University
InventorsChaitan Khosla, Rembert Pieper, Guanglin Luo, David E. Cane, Camilla Kao
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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.
Claims13
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.
3 more elements
Dependencies are inferred from the claim text.