Isopropanol production with engineered Cupriavidus necator as bioproduction platform
Alleviating our society’s dependence on petroleum-based chemicals has been highly emphasized due to fossil fuel shortages and increasing greenhouse gas emissions. Isopropanol is a molecule of high potential to replace some petroleum-based chemicals, which can be produced through biological platforms...
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| Format: | Article |
| Language: | English |
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Springer Berlin Heidelberg
2016
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| Online Access: | http://hdl.handle.net/1721.1/103592 https://orcid.org/0000-0003-0856-0750 https://orcid.org/0000-0002-1015-1270 |
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| author | Grousseau, Estelle Lu, Jingnan Gorret, Nathalie Guillouet, Stéphane E. Sinskey, Anthony J |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Grousseau, Estelle Lu, Jingnan Gorret, Nathalie Guillouet, Stéphane E. Sinskey, Anthony J |
| author_sort | Grousseau, Estelle |
| collection | MIT |
| description | Alleviating our society’s dependence on petroleum-based chemicals has been highly emphasized due to fossil fuel shortages and increasing greenhouse gas emissions. Isopropanol is a molecule of high potential to replace some petroleum-based chemicals, which can be produced through biological platforms from renewable waste carbon streams such as carbohydrates, fatty acids, or CO2. In this study, for the first time, the heterologous expression of engineered isopropanol pathways were evaluated in a Cupriavidus necator strain Re2133, which was incapable of producing poly-3-hydroxybutyrate [P(3HB)]. These synthetic production pathways were rationally designed through codon optimization, gene placement, and gene dosage in order to efficiently divert carbon flow from P(3HB) precursors toward isopropanol. Among the constructed pathways, Re2133/pEG7c overexpressing native C. necator genes encoding a β-ketothiolase, a CoA-transferase, and codon-optimized Clostridium genes encoding an acetoacetate decarboxylase and an alcohol dehydrogenase produced up to 3.44 g l[superscript -1] isopropanol in batch culture, from fructose as a sole carbon source, with only 0.82 g l[superscript -1] of biomass. The intrinsic performance of this strain (maximum specific production rate 0.093 g g[superscript -1] h[superscript -1], yield 0.32 Cmole Cmole[superscript -1]) corresponded to more than 60 % of the respective theoretical performance. Moreover, the overall isopropanol production yield (0.24 Cmole Cmole[superscript -1]) and the overall specific productivity (0.044 g g[superscript -1] h[superscript -1]) were higher than the values reported in the literature to date for heterologously engineered isopropanol production strains in batch culture. Strain Re2133/pEG7c presents good potential for scale-up production of isopropanol from various substrates in high cell density cultures. |
| first_indexed | 2024-09-23T13:41:22Z |
| format | Article |
| id | mit-1721.1/103592 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:41:22Z |
| publishDate | 2016 |
| publisher | Springer Berlin Heidelberg |
| record_format | dspace |
| spelling | mit-1721.1/1035922022-10-01T16:33:09Z Isopropanol production with engineered Cupriavidus necator as bioproduction platform Grousseau, Estelle Lu, Jingnan Gorret, Nathalie Guillouet, Stéphane E. Sinskey, Anthony J Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. Engineering Systems Division Lu, Jingnan Sinskey, Anthony J. Alleviating our society’s dependence on petroleum-based chemicals has been highly emphasized due to fossil fuel shortages and increasing greenhouse gas emissions. Isopropanol is a molecule of high potential to replace some petroleum-based chemicals, which can be produced through biological platforms from renewable waste carbon streams such as carbohydrates, fatty acids, or CO2. In this study, for the first time, the heterologous expression of engineered isopropanol pathways were evaluated in a Cupriavidus necator strain Re2133, which was incapable of producing poly-3-hydroxybutyrate [P(3HB)]. These synthetic production pathways were rationally designed through codon optimization, gene placement, and gene dosage in order to efficiently divert carbon flow from P(3HB) precursors toward isopropanol. Among the constructed pathways, Re2133/pEG7c overexpressing native C. necator genes encoding a β-ketothiolase, a CoA-transferase, and codon-optimized Clostridium genes encoding an acetoacetate decarboxylase and an alcohol dehydrogenase produced up to 3.44 g l[superscript -1] isopropanol in batch culture, from fructose as a sole carbon source, with only 0.82 g l[superscript -1] of biomass. The intrinsic performance of this strain (maximum specific production rate 0.093 g g[superscript -1] h[superscript -1], yield 0.32 Cmole Cmole[superscript -1]) corresponded to more than 60 % of the respective theoretical performance. Moreover, the overall isopropanol production yield (0.24 Cmole Cmole[superscript -1]) and the overall specific productivity (0.044 g g[superscript -1] h[superscript -1]) were higher than the values reported in the literature to date for heterologously engineered isopropanol production strains in batch culture. Strain Re2133/pEG7c presents good potential for scale-up production of isopropanol from various substrates in high cell density cultures. United States. Dept. of Energy MIT-France Seed Fund United States. Advanced Research Projects Agency-Energy France. Ministère de l'éducation nationale, de l'enseignement supérieur et de la recherche (Post-Doctoral grant) Centre National de la Recherche Scientifique (France) 2016-07-13T19:16:33Z 2016-07-13T19:16:33Z 2014-03 2014-02 2016-05-23T12:09:39Z Article http://purl.org/eprint/type/JournalArticle 0175-7598 1432-0614 http://hdl.handle.net/1721.1/103592 Grousseau, Estelle, Jingnan Lu, Nathalie Gorret, Stéphane E. Guillouet, and Anthony J. Sinskey. “Isopropanol Production with Engineered Cupriavidus Necator as Bioproduction Platform.” Applied Microbiology and Biotechnology 98, no. 9 (March 7, 2014): 4277–4290. https://orcid.org/0000-0003-0856-0750 https://orcid.org/0000-0002-1015-1270 en http://dx.doi.org/10.1007/s00253-014-5591-0 Applied Microbiology and Biotechnology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ Springer-Verlag Berlin Heidelberg application/pdf Springer Berlin Heidelberg Springer Berlin Heidelberg |
| spellingShingle | Grousseau, Estelle Lu, Jingnan Gorret, Nathalie Guillouet, Stéphane E. Sinskey, Anthony J Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title | Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title_full | Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title_fullStr | Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title_full_unstemmed | Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title_short | Isopropanol production with engineered Cupriavidus necator as bioproduction platform |
| title_sort | isopropanol production with engineered cupriavidus necator as bioproduction platform |
| url | http://hdl.handle.net/1721.1/103592 https://orcid.org/0000-0003-0856-0750 https://orcid.org/0000-0002-1015-1270 |
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