Method to produce novel polyketides
AssigneesStanford University, Brown University Research Foundation
InventorsChaitan Khosla, Rembert Pieper, Guanglin Luo, David E. Cane, Camilla Kao
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Abstract
Modified PKS gene clusters which produce novel polyketides in an efficient system in a host cell or in a cell free extract are described.
Claims12
Recombinant host cells which contain a polyketide synthase (PKS), gene cluster which encodes a functional modified modular PKS containing at least a first and second module wherein said gene cluster has been modified so that the PKS encoded is unable to utilize a starter unit used by said modular PKS in unmodified form, but is able to incorporate a diketide into at least a triketide,
- said gene cluster operably linked to control sequences for expression,
- wherein at least one of said gene cluster and said control sequences is heterologous to said cells,
- wherein said modification inactivates the ketosynthase (KS) catalytic domain of the first module,
- wherein said inactivation is by modification of a single codon of said catalytic domain,
- wherein said codon, in its unmodified form, encodes cysteine, and
- wherein said codon in its modified form encodes alanine.
A recombinant plasmid vector which comprises an expression system for production of polyketide synthase (PKS) wherein said expression system comprises a nucleotide sequence encoding a functional modified modular PKS operatively linked to control sequences for expression of said modified PKS containing at least a first and second module,
- wherein said nucleotide sequence has been modified so that the PKS encoded is unable to utilize a starter unit used by said modular PKS in unmodified form, but is able to incorporate a diketide into at least a triketide,
- wherein said modification inactivates the ketosynthase (KS) catalytic domain of the first module,
- wherein said inactivation is by modification of a single codon of said catalytic domain,
- wherein said codon, in its unmodified form, encodes cysteine, and
- wherein said codon in its modified form encodes alanine.
Dependencies are inferred from the claim text.