Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol
Increasingly complex metabolic pathways have been engineered by modifying natural pathways and establishing de novo pathways with enzymes from a variety of organisms. Here we apply retro-biosynthetic screening to a modular pathway design to identify a redox neutral, theoretically high yielding route...
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2017
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| Online Access: | http://hdl.handle.net/1721.1/110922 https://orcid.org/0000-0001-6869-9530 https://orcid.org/0000-0003-0437-3157 |
| _version_ | 1826190752804241408 |
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| author | Sheppard, Micah James Kunjapur, Aditya Mohan Wenck, Spencer J. Jones, Kristala L. |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Sheppard, Micah James Kunjapur, Aditya Mohan Wenck, Spencer J. Jones, Kristala L. |
| author_sort | Sheppard, Micah James |
| collection | MIT |
| description | Increasingly complex metabolic pathways have been engineered by modifying natural pathways and establishing de novo pathways with enzymes from a variety of organisms. Here we apply retro-biosynthetic screening to a modular pathway design to identify a redox neutral, theoretically high yielding route to a branched C6 alcohol. Enzymes capable of converting natural E. coli metabolites into 4-methyl-pentanol (4MP) via coenzyme A (CoA)-dependent chemistry were taken from nine different organisms to form a ten-step de novo pathway. Selectivity for 4MP is enhanced through the use of key enzymes acting on acyl-CoA intermediates, a carboxylic acid reductase from Nocardia iowensis and an alcohol dehydrogenase from Leifsonia sp. strain S749. One implementation of the full pathway from glucose demonstrates selective carbon chain extension and acid reduction with 4MP constituting 81% (90±7 mg l⁻¹) of the observed alcohol products. The highest observed 4MP titre is 192±23 mg l⁻¹. These results demonstrate the ability of modular pathway screening to facilitate de novo pathway engineering. |
| first_indexed | 2024-09-23T08:45:10Z |
| format | Article |
| id | mit-1721.1/110922 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:45:10Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
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| spelling | mit-1721.1/1109222022-09-30T11:00:13Z Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol Sheppard, Micah James Kunjapur, Aditya Mohan Wenck, Spencer J. Jones, Kristala L. Massachusetts Institute of Technology. Department of Chemical Engineering Prather, Kristala L. Jones Sheppard, Micah James Kunjapur, Aditya Mohan Wenck, Spencer J. Jones, Kristala L. Increasingly complex metabolic pathways have been engineered by modifying natural pathways and establishing de novo pathways with enzymes from a variety of organisms. Here we apply retro-biosynthetic screening to a modular pathway design to identify a redox neutral, theoretically high yielding route to a branched C6 alcohol. Enzymes capable of converting natural E. coli metabolites into 4-methyl-pentanol (4MP) via coenzyme A (CoA)-dependent chemistry were taken from nine different organisms to form a ten-step de novo pathway. Selectivity for 4MP is enhanced through the use of key enzymes acting on acyl-CoA intermediates, a carboxylic acid reductase from Nocardia iowensis and an alcohol dehydrogenase from Leifsonia sp. strain S749. One implementation of the full pathway from glucose demonstrates selective carbon chain extension and acid reduction with 4MP constituting 81% (90±7 mg l⁻¹) of the observed alcohol products. The highest observed 4MP titre is 192±23 mg l⁻¹. These results demonstrate the ability of modular pathway screening to facilitate de novo pathway engineering. United States. Army Research Office (W911NF-09-0001) 2017-08-04T13:29:23Z 2017-08-04T13:29:23Z 2014-09 2014-03 Article http://purl.org/eprint/type/JournalArticle 2041-1723 http://hdl.handle.net/1721.1/110922 Sheppard, Micah J.; Kunjapur, Aditya M.; Wenck, Spencer J. et al. “Retro-Biosynthetic Screening of a Modular Pathway Design Achieves Selective Route for Microbial Synthesis of 4-Methyl-Pentanol.” Nature Communications 5 (September 2014): 5031. © 2014 Macmillan Publishers Limited https://orcid.org/0000-0001-6869-9530 https://orcid.org/0000-0003-0437-3157 en_US http://dx.doi.org/10.1038/ncomms6031 Nature Communications Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group Prof. Prather via Erja Kajosalo |
| spellingShingle | Sheppard, Micah James Kunjapur, Aditya Mohan Wenck, Spencer J. Jones, Kristala L. Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title | Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title_full | Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title_fullStr | Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title_full_unstemmed | Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title_short | Retro-biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4-methyl-pentanol |
| title_sort | retro biosynthetic screening of a modular pathway design achieves selective route for microbial synthesis of 4 methyl pentanol |
| url | http://hdl.handle.net/1721.1/110922 https://orcid.org/0000-0001-6869-9530 https://orcid.org/0000-0003-0437-3157 |
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