Construction and performance of heterologous polyketide-producing K-12- and B-derived Escherichia coli.
OBJECTIVE
Escherichia coli has emerged as a viable heterologous host for the production of complex, polyketide natural compounds. In this study, polyketide biosynthesis was compared between different E. coli strains for the purpose of better understanding and improving heterologous production.


RESULTS
Both B and K-12 E. coli strains were genetically modified to support heterologous polyketide biosynthesis [specifically, 6-deoxyerythronolide B (6dEB)]. Polyketide production was analysed using a helper plasmid designed to overcome rare codon usage within E. coli. Each strain was analysed for recombinant protein production, precursor consumption, by-product production, and 6dEB biosynthesis. Of the strains tested for biosynthesis, 6dEB production was greatest for E. coli B strains. When comparing biosynthetic improvements as a function of mRNA stability vs codon bias, increased 6dEB titres were observed when additional rare codon tRNA molecules were provided.


CONCLUSIONS
Escherichia coli B strains and the use of tRNA supplementation led to improved 6dEB polyketide titres.


CONCLUSIONS
Given the medicinal potential and growing field of polyketide heterologous biosynthesis, the current study provides insight into host-specific genetic backgrounds and gene expression parameters aiding polyketide production through E. coli.
Wu
J
J
State Key Laboratory of Bioreactor Engineering, National Engineering Research Center for Biotechnology, East China University of Science & Technology, Shanghai, China.
Boghigian
B A
BA
Myint
M
M
Zhang
H
H
Zhang
S
S
Pfeifer
B A
BA
D000900:Anti-Bacterial Agents / Q000378:metabolism; D053898:Biosynthetic Pathways / Q000235:genetics*; D003062:Codon; D004917:Erythromycin / Q000031:analogs & derivatives* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D005818:Genetic Engineering; D006801:Humans; D010957:Plasmids; D020871:RNA Stability; D011994:Recombinant Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic
D000900:Anti-Bacterial Agents; D003062:Codon; D011994:Recombinant Proteins; C034277:6-deoxyerythronolide B; D004917:Erythromycin