DNA encoding methymycin and pikromycin
AssigneeRegents of the University of Minnesota
InventorsDavid H. Sherman, Hung-Wen Liu, Yongquan Xue, Lishan Zhao
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Abstract
A novel pathway for the synthesis of polyhydroxyalkanoates is provided. A method of synthesizing a recombinant polyhydroxyalkanoate monomer synthase is also provided. These recombinant polyhydroxyalkanoate synthases are derived from multifunctional fatty acid synthases or polyketide synthases and generate hydroxyacyl acids capable of polymerization by a polyhydroxyalkanoate synthase. Also provided is a biosynthetic gene cluster for methymycin and pikomycin as well as a biosynthetic gene cluster for desosamine.
Claims26
An isolated and purified nucleic acid segment comprising a nucleic acid sequence comprising a desosamine biosynthetic gene cluster, a fragment or a biologically active variant thereof, wherein the nucleic acid sequence is not derived from the eryC gene cluster of Saccharopolyspora erythraea or Streptomyces antibioticus.
A recombinant bacterial host cell in which at least a portion of a nucleic acid sequence encoding desosamine is disrupted so as to result in a decrease or lack of desosamine synthesis, wherein the nucleic acid sequence which is disrupted is not derived from the eryC gene cluster of Saccharopolyspora erythraea.
An isolated and purified nucleic acid segment comprising a nucleic acid sequence comprising a macrolide biosynthetic gene cluster encoding methymycin, pikomycin, neomethymycin, narbomycin, or a combination thereof, or a biologically active variant or fragment thereof.
An isolated and purified nucleic acid sequence comprising SEQ ID NO:3, SEQ ID NO:5, a fragment thereof, the complement thereto, or which hybridizes thereto.
A recombinant host cell in which a pikAI gene, a pikAII gene, a pikAIII gene, a pikAIV gene, a pikB gene cluster, a pikAV gene cluster, a pikC gene, a pikR1 gene, apikR2 gene, or a combination thereof, is disrupted so as to reduce or eliminate production of methymycin, neomethymycin, pikromycin, narbomycin, or a combination thereof.
An isolated and purified DNA molecule comprising a first DNA segment encoding a first module and a second DNA segment encoding a second module, wherein the DNA segments together encode a recombinant polyhydroxyalkanoate monomer synthase, and wherein at least one DNA segment is derived from the pikA gene cluster of Streptomyces venezuelae.
A method of providing a polyhydroxyalkanoate monomer, comprising:
- (a) introducing into a host cell a DNA molecule comprising a DNA segment encoding a recombinant polyhydroxyalkanoate monomer synthase operably linked to a promoter functional in the host cell, wherein the recombinant polyhydroxyalkanoate monomer synthase comprises a first module and a second module, and wherein at least one DNA segment is derived from the pikA gene cluster of Streptomyces venezuelae;
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- (b) expressing the DNA encoding the recombinant polyhydroxyalkanoate monomer synthase in the host cell so as to generate a polyhydroxyalkanoate monomer.
A recombinant vector comprising one or more modules of a polyketide synthase wherein at least one module is from Streptomyces venezuelae.
A method of providing a polyhydroxyalkanoate monomer, comprising:
- (a) introducing into a host cell a DNA molecule encoding a fusion polypeptide, wherein the DNA molecule comprises a first DNA segment operably linked to a promoter functional in the host cell and a second DNA segment, wherein at least one DNA segment is derived from the pikA gene cluster of Streptomyces venezuelae;
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- (b) expressing the DNA in the host cell so as to generate the fusion polypeptide.
Dependencies are inferred from the claim text.