期刊
BMC BIOTECHNOLOGY
卷 13, 期 -, 页码 -出版社
BMC
DOI: 10.1186/1472-6750-13-6
关键词
Lycopene; Escherichia coli; Chemically induced chromosomal evolution; Metabolic engineering
资金
- National Natural Science Foundation of China [30970089, 20876181, 21276289]
- Natural Science Foundation of Guangdong Province [9351027501000003, S2011010001396]
Background: Plasmid-based overexpression of genes has been the principal strategy for metabolic engineering. However, for biotechnological applications, plasmid-based expression systems are not suitable because of genetic instability, and the requirement for constant selective pressure to ensure plasmid maintenance. Results: To overcome these drawbacks, we constructed an Escherichia coli lycopene production strain that does not carry a plasmid or an antibiotic marker. This was achieved using triclosan-induced chromosomal evolution, a high gene copy expression system. The engineered strain demonstrated high genetic stability in the absence of the selective agent during fermentation. The replacement of native appY promoter with a T5 promoter, and the deletion of the iclR gene in E. coli CBW 12241 further improved lycopene production. The resulting strain, E. coli CBW 12241(Delta iclR, P-T5-appY), produced lycopene at 33.43 mg per gram of dry cell weight. Conclusions: A lycopene hyper-producer E. coli strain that does not carry a plasmid or antibiotic marker was constructed using triclosan-induced chromosomal evolution. The methods detailed in this study can be used to engineer E. coli to produce other metabolites.
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