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A highly selective biosynthetic pathway to non-natural C50 carotenoids assembled from moderately selective enzymes

Author

Listed:
  • Maiko Furubayashi

    (Chiba University
    Present address: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA)

  • Mayu Ikezumi

    (Chiba University)

  • Shinichi Takaichi

    (Nippon Medical School)

  • Takashi Maoka

    (Research Institute for Production Development)

  • Hisashi Hemmi

    (Nagoya University)

  • Takuya Ogawa

    (Nagoya University)

  • Kyoichi Saito

    (Chiba University)

  • Alexander V Tobias

    (DuPont Industrial Biosciences, Experimental Station)

  • Daisuke Umeno

    (Chiba University
    Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST))

Abstract

Synthetic biology aspires to construct natural and non-natural pathways to useful compounds. However, pathways that rely on multiple promiscuous enzymes may branch, which might preclude selective production of the target compound. Here, we describe the assembly of a six-enzyme pathway in Escherichia coli for the synthesis of C50-astaxanthin, a non-natural purple carotenoid. We show that by judicious matching of engineered size-selectivity variants of the first two enzymes in the pathway, farnesyl diphosphate synthase (FDS) and carotenoid synthase (CrtM), branching and the production of non-target compounds can be suppressed, enriching the proportion of C50 backbones produced. We then further extend the C50 pathway using evolved or wild-type downstream enzymes. Despite not containing any substrate- or product-specific enzymes, the resulting pathway detectably produces only C50 carotenoids, including ∼90% C50-astaxanthin. Using this approach, highly selective pathways can be engineered without developing absolutely specific enzymes.

Suggested Citation

  • Maiko Furubayashi & Mayu Ikezumi & Shinichi Takaichi & Takashi Maoka & Hisashi Hemmi & Takuya Ogawa & Kyoichi Saito & Alexander V Tobias & Daisuke Umeno, 2015. "A highly selective biosynthetic pathway to non-natural C50 carotenoids assembled from moderately selective enzymes," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8534
    DOI: 10.1038/ncomms8534
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