Metabolic engineering of sesquiterpene metabolism in yeast

Authors

  • Shunji Takahashi,

    Corresponding author
    1. Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
    Current affiliation:
    1. Antibiotics Laboratory, Discovery Research Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
    • Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
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  • Yunsoo Yeo,

    1. Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
    Current affiliation:
    1. Microbial Genetics Division, National Institute of Agricultural Biotechnology, Suwon 441-707, Korea.
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  • Bryan T. Greenhagen,

    1. Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
    Current affiliation:
    1. Allylix, Inc, A165 ASTeCC, University of Kentucky, Lexington, Kentucky 40546.
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  • Tom McMullin,

    1. Bio-Technical Resources, 1035 South 7th Street, Manitowoc, Wisconsin 54220
    Current affiliation:
    1. NatureWorks LLC, 15305 Minnetonka Blvd., Minnetonka, Minnesota 55345.
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  • Linsheng Song,

    1. Bio-Technical Resources, 1035 South 7th Street, Manitowoc, Wisconsin 54220
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  • Julie Maurina-Brunker,

    1. Bio-Technical Resources, 1035 South 7th Street, Manitowoc, Wisconsin 54220
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  • Reinhardt Rosson,

    1. Bio-Technical Resources, 1035 South 7th Street, Manitowoc, Wisconsin 54220
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  • Joseph P. Noel,

    1. Howard Hughes Medical Institute, The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, La Jolla, California 92037
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  • Joe Chappell

    Corresponding author
    1. Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
    • Department of Plant & Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312; telephone: (859)257-5020; fax: (859)257-7125
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Abstract

Terpenes are structurally diverse compounds that are of interest because of their biological activities and industrial value. These compounds consist of chirally rich hydrocarbon backbones derived from terpene synthases, which are subsequently decorated with hydroxyl substituents catalyzed by terpene hydroxylases. Availability of these compounds is, however, limited by intractable synthetic means and because they are produced in low amounts and as complex mixtures by natural sources. We engineered yeast for sesquiterpene accumulation by introducing genetic modifications that enable the yeast to accumulate high levels of the key intermediate farnesyl diphosphate (FPP). Co-expression of terpene synthase genes diverted the enlarged FPP pool to greater than 80 mg/L of sesquiterpene. Efficient coupling of terpene production with hydroxylation was also demonstrated by coordinate expression of terpene hydroxylase activity, yielding 50 mg/L each of hydrocarbon and hydroxylated products. These yeast now provide a convenient format for investigating catalytic coupling between terpene synthases and hydroxylases, as well as a platform for the industrial production of high value, single-entity and stereochemically unique terpenes. Biotechnol. Bioeng. 2007;97: 170–181. © 2006 Wiley Periodicals, Inc.

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