Heterologous expression of epothilone biosynthetic genes in Myxococcus xanthus.
Epothilones are potential anticancer drugs that stabilize microtubules in a manner similar to paclitaxel (Taxol). Epothilones are produced from the myxobacterium Sorangium cellulosum, which has a 16-h doubling time and produces only milligram-per-liter amounts of epothilone A and epothilone B. Furthermore, genetic manipulation of S. cellulosum is difficult. To produce epothilones in a more genetically amenable and rapidly growing host, we chose the closely related and best-characterized myxobacteria Myxococcus xanthus. We inserted 65.4 kb of S. cellulosum DNA that encompassed the entire epothilone gene cluster into the chromosome of M. xanthus by a series of homologous recombination events. The resulting strain produced epothilones A and B. Construction of a strain that contained a mutation in epoK, the P450 epoxidase, resulted in production of epothilones C and D.
Julien
Bryan
B
Kosan Biosciences, Inc., Hayward, California 94545, USA. julien@kosan.com
Shah
Sanjay
S
D001426:Bacterial Proteins / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics; D003470:Culture Media; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D034261:Epothilones / Q000096:biosynthesis* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D005810:Multigene Family / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016941:Myxococcus xanthus / Q000235:genetics* / Q000378:metabolism*; D010088:Oxidoreductases / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering
D001426:Bacterial Proteins; D003470:Culture Media; D004269:DNA, Bacterial; D034261:Epothilones; C426624:epothilone C; D003577:Cytochrome P-450 Enzyme System; C403885:EpoK protein, Polyangium cellulosum; D010088:Oxidoreductases; C114026:desoxyepothilone B