High Isoprenoid Flux Escherichia coli as a Host for Carotenoids Production
A noncarotenogenic microbe E. coli was engineered for high production of carotenoids. To increase the isoprenoid flux, the chromosomal native promoters of the rate-controlling steps (dxs, idi and ispDispF) in the isoprenoid pathway were replaced with a strong bacteriophage T5 promoter (PT5) by using the λ-Red recombinase system in combination with the Flp/FRT site-specific recombination system for marker excision and P1 transduction for gene trait stacking. The resulting high isoprenoid flux E. coli can be used as a starting strain to produce various carotenoids by introducing heterologous carotenoid genes. In this study, the high isoprenoid flux E. coli was transformed with a plasmid carrying the β-carotene biosynthetic genes from Pantoea stewartii for β-carotene production.
Key wordsHigh isoprenoid flux Escherichia coli Chromosomal promoter replacement Metabolic engineering Isoprenoid pathway Carotenoids β-carotene
The author thanks Professor Barry L. Wanner for providing λ-Red strains and plasmids used in this work and Wendy Suh for critical reading of the manuscript.
- 3.Rohdich, F., Zepeck, F., Adam, P., Hecht, S., Kaiser, J., Laupitz, R., Gräwert, T., Amslinger, S., Eisenreich, W., Bacher, A., and Arigoni, D. (2003) The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: Studies on the mechanisms of the reactions catalyzed by IspG and IspH protein. Proc. Natl. Acad. Sci. USA. 100, 1586–1591.PubMedCrossRefGoogle Scholar
- 12.Bachmann, B. J. (1996) Derivations and genotypes of some mutant derivatives of Escherichia coli K-12. In “Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology” (F. C. Neidhardt, R. Curtiss III, J. L. Ingraham, E. C. C. Lin, K. B. Low, B. Magasanik, W. S. Reznikoff, M. Riley, M. Schaechter, and H. E. Umbarger, Eds.), 2nd ed., pp. 2460–2488, ASM Press, Washington, DC.Google Scholar
- 13.Miller, J. H. (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar