Precursor-directed biosynthesis of epothilone in Escherichia coli.
Engineered biosynthetic pathways provide a powerful method for generating complex molecules. Precursor-directed biosynthesis, which combines chemical synthesis and enzymatic transformations, allows non-native starting materials to be incorporated into biosynthetic pathways. Using this approach, we achieved the production of the anticancer agent epothilone C in Escherichia coli. An E. coli strain was engineered to express the last three modules of the epothilone biosynthetic pathway (epoD-M6, epoE, and epoF) and the substrate required to complement the biosynthetic enzymes was obtained by chemical synthesis. Under high-density cell culture conditions, the E. coli strain processed exogenously fed synthetic substrate into epothilone C at levels comparable to the native host (1 mg/L) and at higher levels than other heterologous hosts. Importantly, this precursor-directed approach will allow chemical modifications to be introduced into the polyketide backbone and may ultimately provide access to epothilone analogues with improved pharmacological properties in quantities sufficient for clinical development.
Boddy
Christopher N
CN
Departments of Chemical Engineering, Chemistry, and Biochemistry, Stanford University, Stanford, California 94305-5025, USA.
Hotta
Kinya
K
Tse
Martha Lovato
ML
Watts
R Edward
RE
Khosla
Chaitan
C
D000903:Antibiotics, Antineoplastic / Q000032:analysis / Q000096:biosynthesis; D034261:Epothilones / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000235:genetics; D011498:Protein Precursors / Q000502:physiology*
D000903:Antibiotics, Antineoplastic; D034261:Epothilones; D009097:Multienzyme Complexes; D011498:Protein Precursors; C426624:epothilone C