Recombinant methods and materials for producing epothilone and epothilone derivatives
AssigneeKosan Biosciences, Inc.
InventorsBryan Julien, Leonard Katz, Chaitan Khosla, Li Tang, Rainer Ziermann
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Abstract
Recombinant nucleic acids that encode all or a portion of the epothilone polyketide synthase (PKS) are used to express recombinant PKS genes in host cells for the production of epothilones, epothilone derivatives, and polyketides that are useful as cancer chemotherapeutics, fungicides, and immunosuppressants.
Claims10
A recombinant Sorangium cellulosum host cell that contains at least one mutated gene for an epothilone PKS protein or epothilone modification enzyme, wherein said mutated gene is inserted in whole or in part into the genomic DNA of said cell by homologous recombination with a recombinant vector comprising all or a part of a mutated epothilone epoK gene, wherein said recombinant host cell produces more epothilone C than epothilone A and/or more epothilone D than epothilone B.
A recombinant Sorangium cellulosum host cell that contains a mutated epoK gene that does not produce an enzymatically active epoK gene product, wherein said cell produces more epothilone C than epothilone A and/or more epothilone D than epothilone B.
An isolated or recombinant host cell that contains a nucleic acid that codes for expression of Sorangium cellulosum epothilone polyketide synthase (“PKS”), wherein said PKS contains an altered epoD extender module 4 acyltransferase (“AT”) domain, whereby said acyltransferase domain has specificity for either malonyl CoA or for methylmalonyl CoA, but does not contain an enzymatically active product of a Sorangium cellulosum epoK gene.
A recombinant Streptomyces or Myxococcus host cell that expresses Sorangium cellulosum epothilone polyketide synthase (“PKS”) genes, said genes including an epoD gene that comprises an altered extender module 4 acyltransferase (“AT”) domain coding sequence, whereby said acyltransferase domain has specificity for either malonyl CoA or for methylmalonyl CoA, and the cell does not express an enzymatically active Sorangium cellulosum epoK gene product, wherein said Strentomyces or Myxococcus host cell optionally comprises one or more of said epothilone PKS genes, either integrated into its chromosomal DNA or comprised by an extrachromosomal expression vector, wherein said recombinant Streptomyces or Myxococcus host cell produces more epothilone C than epothilone A and/or more epothilone D than epothilone B.
Dependencies are inferred from the claim text.