A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes
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Furubayashi-2015-Highly selective.pdf
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Author(s)
Furubayashi, Maiko
Ikezumi, Mayu
Takaichi, Shinichi
Maoka, Takashi
Hemmi, Hisashi
Ogawa, Takuya
Saito, Kyoichi
Tobias, Alexander V
Umeno, Daisuke
Date Issued
July 2015
Journal
Nature Communications
Publisher
Nature Publishing Group
Citation
Furubayashi, Maiko, Mayu Ikezumi, Shinichi Takaichi, Takashi Maoka, Hisashi Hemmi, Takuya Ogawa, Kyoichi Saito, Alexander V Tobias, and Daisuke Umeno. “A Highly Selective Biosynthetic Pathway to Non-Natural C[subscript 50] Carotenoids Assembled from Moderately Selective Enzymes.” Nature Communications 6 (July 14, 2015): 7534. © 2015 Macmillan Publishers Limited
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Final published version
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 C[subscript 50]-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 C[subscript 50] backbones produced. We then further extend the C[subscript 50] pathway using evolved or wild-type downstream enzymes. Despite not containing any substrate- or product-specific enzymes, the resulting pathway detectably produces only C[subscript 50] carotenoids, including ~90% C[subscript 50]-astaxanthin. Using this approach, highly selective pathways can be engineered without developing absolutely specific enzymes.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
https://doi.org/10.1038/ncomms8534
