Heterologous Production of Epothilone C and D in Escherichia coli

The epothilones are a family of polyketide natural products that show a high potential as anticancer drugs. They are synthesized by the action of a hybrid nonribosomal peptide synthetase/polyketide synthase in the myxobacterium Sorangium cellulosum. In this work, the genes encoding the entire cluste...

Full description

Saved in:
Bibliographic Details
Published in:Biochemistry (Easton) 2006-01, Vol.45 (4), p.1321-1330
Main Authors: Mutka, Sarah C, Carney, John R, Liu, Yaoquan, Kennedy, Jonathan
Format: Article
Language:English
Subjects:
Catalysis
Cysteamine - analogs & derivatives
Cysteamine - metabolism
Epothilones - biosynthesis
Epothilones - metabolism
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Methacrylates - metabolism
Peptide Chain Elongation, Translational - genetics
Peptide Chain Elongation, Translational - physiology
Peptide Synthases - genetics
Peptide Synthases - metabolism
Polyketide Synthases - biosynthesis
Polyketide Synthases - genetics
Polyketide Synthases - metabolism
Sorangium cellulosum
Thiazoles - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The epothilones are a family of polyketide natural products that show a high potential as anticancer drugs. They are synthesized by the action of a hybrid nonribosomal peptide synthetase/polyketide synthase in the myxobacterium Sorangium cellulosum. In this work, the genes encoding the entire cluster, epoA, epoB, epoC, epoD, epoE, and epoF, were redesigned and synthesized to allow for expression in Escherichia coli. The expression of the largest of the proteins, EpoD, also required the protein be separated into two polypeptides with compatible module linkers. Using a combination of lowered temperature, chaperone coexpression, and alternative promoters, we succeeded in producing a soluble protein from all genes in the epothilone cluster. The entire synthetic epothilone cluster was then expressed in a strain of E. coli modified to enable polyketide biosynthesis, resulting in the production of epothilones C and D. Furthermore, feeding a thioester of the normal substrate for EpoD to cells expressing the epoD, epoE, and epoF genes also led to the production of epothilones C and D. The design of the synthetic epothilone genes together with E. coli expression provides the ideal platform for both the biochemical investigation of the epothilone PKS and the generation of novel biosynthetic epothilone analogues.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi052075r