Recombinant methods and materials for producing epothilones C and D
AssigneeKason Biosciences, Inc.
InventorsBryan Julien, Leonard Katz, Chaitan Khosla, Li Tang
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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.
Claims29
A method to produce epothilone C and/or epothilone D, which method comprises culturing an isolated or recombinant host cell that contains a nucleic acid that codes for expression of Sorangium cellulosum epothilone polyketide synthase (“PKS”) but does not contain a functionally active product of a Sorangium cellulosum epoK gene under conditions wherein the nucleic acid is expressed to obtain a functional PKS, and epothilone C and/or epothilone D are produced.
A method to produce epothilone C and/or epothilone D, which method comprises culturing 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 but does not contain a functionally active product of a Sorangium cellulosum epoK gene under conditions wherein the nucleic acid is expressed to obtain a functional PKS, and epothilone C and/or epothilone D are produced.
A method to produce epothilone C and/or epothilone D, which comprises culturing recombinant Streptomyces or Myxococcus host cells that express Sorangium cellulosum epothilone polyketide synthase (“PKS”) genes and do not express a fully functional Sorangium cellulosum epoK gene product, optionally comprising one or more of said epothilone PKS genes integrated into their chromosomal DNA or on an extrachromosomal expression vector, wherein said recombinant cells produce more epothilone C than epothilone A and/or more epothilone D than epothilone B.
A method to produce epothilone C and/or epothilone D, which comprises culturing recombinant Streptomyces or Myxococcus host cells that express 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, and do not express a fully functional Sorangium cellulosum epoK gene product, optionally comprising one or more of said epothilone PKS genes integrated into their chromosomal DNA or on an extrachromosomal expression vector, wherein said recombinant cells produce more epothilone C than epothilone A and/or more epothilone D than epothilone B.
A method to produce epothilone C and/or epothilone D, which method comprises culturing a recombinant Sorangium cellulosum host cell that contains a mutated epoK gene that does not produce a fully functional gene product, wherein said cell produces more epothilone C than epothilone A and/or more epothilone D than epothilone B.
Dependencies are inferred from the claim text.