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Chromosomal evolution of Escherichia coli for the efficient production of lycopene

Author:
Chen, Yun-Yan  Shen, Hong-Jie  Cui, Yan-Yan  Chen, Shang-Guang  Weng, Zhi-Ming  Zhao, Ming  Liu, Jian-Zhong  


Journal:
BMC BIOTECHNOLOGY


Issue Date:
2013


Abstract(summary):

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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