The biosynthetic gene cluster for the microtubule-stabilizing agents epothilones A and B from Sorangium cellulosum So ce90.
BACKGROUND
Epothilones are produced by the myxobacterium Sorangium cellulosum So ce90, and, like paclitaxel (Taxol((R))), they inhibit microtubule depolymerisation and arrest the cell cycle at the G2-M phase. They are effective against P-glycoprotein-expressing multiple-drug-resistant tumor cell lines and are more water soluble than paclitaxel. The total synthesis of epothilones has been achieved, but has not provided an economically viable alternative to fermentation. We set out to clone, sequence and analyze the gene cluster responsible for the biosynthesis of the epothilones in S. cellulosum So ce90.


RESULTS
A cluster of 22 open reading frames spanning 68,750 base pairs of the S. cellulosum So ce90 genome has been sequenced and found to encode nine modules of a polyketide synthase (PKS), one module of a nonribosomal peptide synthetase (NRPS), a cytochrome P450, and two putative antibiotic transport proteins. Disruptions in the genes encoding the PKS abolished epothilone production. The first PKS module and the NRPS module are proposed to co-operate in forming the thiazole heterocycle of epothilone from an acetate and a cysteine by condensation, cyclodehydration and subsequent dehydrogenation. The remaining eight PKS modules are responsible for the elaboration of the rest of the epothilone carbon skeleton.


CONCLUSIONS
The overall architecture of the gene cluster responsible for epothilone biosynthesis has been determined. The availability of the cluster should facilitate the generation of designer epothilones by combinatorial biosynthesis approaches, and the heterologous expression of epothilones in surrogate microbial hosts.
Molnár
I
I
Novartis Agribusiness Biotechnology Research, Inc., PO Box 12257, NC 27709, USA. istvan.molnar@nabri.novartis.com
Schupp
T
T
Ono
M
M
Zirkle
R
R
Milnamow
M
M
Nowak-Thompson
B
B
Engel
N
N
Toupet
C
C
Stratmann
A
A
Cyr
D D
DD
Gorlach
J
J
Mayo
J M
JM
Hu
A
A
Goff
S
S
Schmid
J
J
Ligon
J M
JM
D000900:Anti-Bacterial Agents; D000970:Antineoplastic Agents / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D034261:Epothilones; D004852:Epoxy Compounds / Q000378:metabolism*; D015723:Gene Library; D005798:Genes, Bacterial; D018942:Macrolides; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics; D005810:Multigene Family / Q000235:genetics*; D009231:Myxococcales / Q000737:chemistry* / Q000235:genetics*; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D013844:Thiazoles / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D034261:Epothilones; D004852:Epoxy Compounds; D018942:Macrolides; D009097:Multienzyme Complexes; D013844:Thiazoles; C093787:epothilone A; D010453:Peptide Synthases; C093788:epothilone B