Isolation and characterization of the epothilone biosynthetic gene cluster from Sorangium cellulosum.
The epothilone biosynthetic gene cluster was isolated from Sorangium cellulosum strain SMP44. The gene cluster contains seven genes and spans approx. 56kb. The genes encoding the PKS, epoA, epoC, epoD, epoE, and epoF, are divided into nine modules. The EpoB protein is a non-ribosomal peptide synthetase (NRPS) that catalyzes formation of the thiazole found in the epothilones. EpoK is a P450 enzyme responsible for the epoxidation of epothilones C and D to epothilones A and B, respectively. EpoK was expressed in Escherichia coli, and the purified protein was shown to convert epothilone D to epothilone B in vitro.
Julien
B
B
Kosan Biosciences, Inc., Hayward, CA 94545, USA. julien@kosan.com
Shah
S
S
Ziermann
R
R
Goldman
R
R
Katz
L
L
Khosla
C
C
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D034261:Epothilones; D004852:Epoxy Compounds / Q000378:metabolism*; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D009231:Myxococcales / Q000235:genetics* / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013844:Thiazoles / Q000378:metabolism*
D001426:Bacterial Proteins; D034261:Epothilones; D004852:Epoxy Compounds; D009097:Multienzyme Complexes; D013844:Thiazoles; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; C403885:EpoK protein, Polyangium cellulosum; D010088:Oxidoreductases; C093788:epothilone B