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Molecular breeding of carotenoid biosynthetic pathways

Nature Biotechnologyvolume 18pages750753 (2000) | Download Citation

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Abstract

The burgeoning demand for complex, biologically active molecules for medicine, materials science, consumer products, and agrochemicals is driving efforts to engineer new biosynthetic pathways into microorganisms and plants. We have applied principles of breeding, including mixing genes and modifying catalytic functions by in vitro evolution, to create new metabolic pathways for biosynthesis of natural products in Escherichia coli. We expressed shuffled phytoene desaturases in the context of a carotenoid biosynthetic pathway assembled from different bacterial species and screened the resulting library for novel carotenoids. One desaturase chimera efficiently introduced six rather than four double bonds into phytoene, to favor production of the fully conjugated carotenoid, 3,4,3′,4′-tetradehydrolycopene. This new pathway was extended with a second library of shuffled lycopene cyclases to produce a variety of colored products. One of the new pathways generates the cyclic carotenoid torulene, for the first time, in E. coli. This combined approach of rational pathway assembly and molecular breeding may allow the discovery and production, in simple laboratory organisms, of new compounds that are essentially inaccessible from natural sources or by synthetic chemistry.

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Acknowledgements

We are grateful to Dr. Peter G. Green (Environmental Analysis Center, Caltech) for technical support and helpful discussions on mass analysis. C.S.-D. was supported by the Deutsche Forschungsgemeinschaft (DFG) and D.U. by the Japan Society for the Promotion of Science (JSPS).

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  1. Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, St. Paul, 55108, MN

    • Claudia Schmidt-Dannert
  2. Division of Chemistry and Chemical Engineering 210-41, California Institute of Technology, Pasadena, 91125, CA

    • Daisuke Umeno
    •  & Frances H. Arnold

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Correspondence to Claudia Schmidt-Dannert or Frances H. Arnold.

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https://doi.org/10.1038/77319

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