Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source
3-hydroxy-γ-butyrolactone (3HBL) is a versatile chiral synthon, deemed a top value-added chemical from biomass by the DOE. We recently reported the first biosynthetic pathway towards 3HBL and its hydrolyzed form, 3,4-dihydroxybutyric acid (3,4-DHBA) in recombinant Escherichia coli using glucose and...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2016
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| Online Access: | http://hdl.handle.net/1721.1/101387 https://orcid.org/0000-0002-4593-5606 https://orcid.org/0000-0003-0437-3157 |
| _version_ | 1811093613259522048 |
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| author | Tarasova, Yekaterina Martin, Collin H. Dhamankar, Himanshu Hemant Prather, Kristala L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Tarasova, Yekaterina Martin, Collin H. Dhamankar, Himanshu Hemant Prather, Kristala L |
| author_sort | Tarasova, Yekaterina |
| collection | MIT |
| description | 3-hydroxy-γ-butyrolactone (3HBL) is a versatile chiral synthon, deemed a top value-added chemical from biomass by the DOE. We recently reported the first biosynthetic pathway towards 3HBL and its hydrolyzed form, 3,4-dihydroxybutyric acid (3,4-DHBA) in recombinant Escherichia coli using glucose and glycolic acid as feedstocks and briefly described their synthesis solely from glucose. Synthesis from glucose requires integration of the endogenous glyoxylate shunt with the 3,4-DHBA/3HBL pathway and co-overexpression of seven genes, posing challenges with respect to expression, repression of the glyoxylate shunt and optimal carbon distribution between the two pathways. Here we discuss engineering this integration. While appropriate media and over-expression of glyoxylate shunt enzymes helped overcome repression, two orthogonal expression systems were employed to address the expression and carbon distribution challenge. Synthesis of up to 0.3 g/L of 3HBL and 0.7 g/L of 3,4-DHBA solely from glucose was demonstrated, amounting to 24% of the theoretical maximum. |
| first_indexed | 2024-09-23T15:47:53Z |
| format | Article |
| id | mit-1721.1/101387 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:47:53Z |
| publishDate | 2016 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1013872022-10-02T04:09:54Z Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source Tarasova, Yekaterina Martin, Collin H. Dhamankar, Himanshu Hemant Prather, Kristala L Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Microbiology Graduate Program Prather, Kristala L. Jones Dhamankar, Himanshu Tarasova, Yekaterina Martin, Collin H. Prather, Kristala L. Jones 3-hydroxy-γ-butyrolactone (3HBL) is a versatile chiral synthon, deemed a top value-added chemical from biomass by the DOE. We recently reported the first biosynthetic pathway towards 3HBL and its hydrolyzed form, 3,4-dihydroxybutyric acid (3,4-DHBA) in recombinant Escherichia coli using glucose and glycolic acid as feedstocks and briefly described their synthesis solely from glucose. Synthesis from glucose requires integration of the endogenous glyoxylate shunt with the 3,4-DHBA/3HBL pathway and co-overexpression of seven genes, posing challenges with respect to expression, repression of the glyoxylate shunt and optimal carbon distribution between the two pathways. Here we discuss engineering this integration. While appropriate media and over-expression of glyoxylate shunt enzymes helped overcome repression, two orthogonal expression systems were employed to address the expression and carbon distribution challenge. Synthesis of up to 0.3 g/L of 3HBL and 0.7 g/L of 3,4-DHBA solely from glucose was demonstrated, amounting to 24% of the theoretical maximum. National Science Foundation (U.S.). Synthetic Biology Engineering Research Center (Grant EEC-0540879) Masdar Institute of Science and Technology (Massachusetts Institute of Technology Cooperative Agreement 02/MI/MI/CP/11/07633/GEN/G/00) 2016-03-01T20:39:43Z 2016-03-01T20:39:43Z 2014-06 2014-04 Article http://purl.org/eprint/type/JournalArticle 10967176 1096-7184 http://hdl.handle.net/1721.1/101387 Dhamankar, Himanshu, Yekaterina Tarasova, Collin H. Martin, and Kristala L.J. Prather. “Engineering E. Coli for the Biosynthesis of 3-Hydroxy-γ-Butyrolactone (3HBL) and 3,4-Dihydroxybutyric Acid (3,4-DHBA) as Value-Added Chemicals from Glucose as a Sole Carbon Source.” Metabolic Engineering 25 (September 2014): 72–81. https://orcid.org/0000-0002-4593-5606 https://orcid.org/0000-0003-0437-3157 en_US http://dx.doi.org/10.1016/j.ymben.2014.06.004 Metabolic Engineering Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier Prof. Prather via Erja Kajosalo |
| spellingShingle | Tarasova, Yekaterina Martin, Collin H. Dhamankar, Himanshu Hemant Prather, Kristala L Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title | Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title_full | Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title_fullStr | Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title_full_unstemmed | Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title_short | Engineering E. coli for the biosynthesis of 3-hydroxy-γ-butyrolactone (3HBL) and 3,4-dihydroxybutyric acid (3,4-DHBA) as value-added chemicals from glucose as a sole carbon source |
| title_sort | engineering e coli for the biosynthesis of 3 hydroxy γ butyrolactone 3hbl and 3 4 dihydroxybutyric acid 3 4 dhba as value added chemicals from glucose as a sole carbon source |
| url | http://hdl.handle.net/1721.1/101387 https://orcid.org/0000-0002-4593-5606 https://orcid.org/0000-0003-0437-3157 |
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