A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes
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...
| Main Authors: | , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Nature Publishing Group
2015
|
| Online Access: | http://hdl.handle.net/1721.1/98477 https://orcid.org/0000-0001-5215-523X |
| _version_ | 1826190541631520768 |
|---|---|
| author | Furubayashi, Maiko Ikezumi, Mayu Takaichi, Shinichi Maoka, Takashi Hemmi, Hisashi Ogawa, Takuya Saito, Kyoichi Tobias, Alexander V Umeno, Daisuke |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Furubayashi, Maiko Ikezumi, Mayu Takaichi, Shinichi Maoka, Takashi Hemmi, Hisashi Ogawa, Takuya Saito, Kyoichi Tobias, Alexander V Umeno, Daisuke |
| author_sort | Furubayashi, Maiko |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T08:41:52Z |
| format | Article |
| id | mit-1721.1/98477 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:41:52Z |
| publishDate | 2015 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/984772022-09-30T10:34:27Z A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes Furubayashi, Maiko Ikezumi, Mayu Takaichi, Shinichi Maoka, Takashi Hemmi, Hisashi Ogawa, Takuya Saito, Kyoichi Tobias, Alexander V Umeno, Daisuke Massachusetts Institute of Technology. Department of Biological Engineering Furubayashi, Maiko 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. Japan Society for the Promotion of Science (Fellowship for Young Scientists) 2015-09-14T13:47:54Z 2015-09-14T13:47:54Z 2015-07 2015-02 Article http://purl.org/eprint/type/JournalArticle 2041-1723 http://hdl.handle.net/1721.1/98477 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 https://orcid.org/0000-0001-5215-523X en_US http://dx.doi.org/10.1038/ncomms8534 Nature Communications Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group Nature Publishing Group |
| spellingShingle | Furubayashi, Maiko Ikezumi, Mayu Takaichi, Shinichi Maoka, Takashi Hemmi, Hisashi Ogawa, Takuya Saito, Kyoichi Tobias, Alexander V Umeno, Daisuke A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title | A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title_full | A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title_fullStr | A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title_full_unstemmed | A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title_short | A highly selective biosynthetic pathway to non-natural C[subscript 50] carotenoids assembled from moderately selective enzymes |
| title_sort | highly selective biosynthetic pathway to non natural c subscript 50 carotenoids assembled from moderately selective enzymes |
| url | http://hdl.handle.net/1721.1/98477 https://orcid.org/0000-0001-5215-523X |
| work_keys_str_mv | AT furubayashimaiko ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT ikezumimayu ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT takaichishinichi ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT maokatakashi ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT hemmihisashi ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT ogawatakuya ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT saitokyoichi ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT tobiasalexanderv ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT umenodaisuke ahighlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT furubayashimaiko highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT ikezumimayu highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT takaichishinichi highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT maokatakashi highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT hemmihisashi highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT ogawatakuya highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT saitokyoichi highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT tobiasalexanderv highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes AT umenodaisuke highlyselectivebiosyntheticpathwaytononnaturalcsubscript50carotenoidsassembledfrommoderatelyselectiveenzymes |