Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds
© 2020 Engineering microbes to utilize non-conventional substrates could create short and efficient pathways to convert substrate into product. In this study, we designed and constructed a two-step heterologous ethanol utilization pathway (EUP) in Escherichia coli by using acetaldehyde dehydrogenase...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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Elsevier BV
2022
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| Online Access: | https://hdl.handle.net/1721.1/133403.2 |
| _version_ | 1811068345375522816 |
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| author | Liang, Hong Ma, Xiaoqiang Ning, Wenbo Liu, Yurou Sinksey, Anthony J. Stephanopoulos, Gregory Zhou, Kang |
| author2 | Singapore-MIT Alliance in Research and Technology (SMART) |
| author_facet | Singapore-MIT Alliance in Research and Technology (SMART) Liang, Hong Ma, Xiaoqiang Ning, Wenbo Liu, Yurou Sinksey, Anthony J. Stephanopoulos, Gregory Zhou, Kang |
| author_sort | Liang, Hong |
| collection | MIT |
| description | © 2020 Engineering microbes to utilize non-conventional substrates could create short and efficient pathways to convert substrate into product. In this study, we designed and constructed a two-step heterologous ethanol utilization pathway (EUP) in Escherichia coli by using acetaldehyde dehydrogenase (encoded by ada) from Dickeya zeae and alcohol dehydrogenase (encoded by adh2) from Saccharomyces cerevisiae. This EUP can convert ethanol into acetyl-CoA without ATP consumption, and generate two molecules of NADH per molecule of ethanol. We optimized the expression of these two genes and found that ethanol consumption could be improved by expressing them in a specific order (ada-adh2) with a constitutive promoter (PgyrA). The engineered E. coli strain with EUP consumed approximately 8 g/L of ethanol in 96 h when it was used as sole carbon source. Subsequently, we combined EUP with the biosynthesis of polyhydroxybutyrate (PHB), a biodegradable polymer derived from acetyl-CoA. The engineered E. coli strain carrying EUP and PHB biosynthetic pathway produced 1.1 g/L of PHB from 10 g/L of ethanol and 1 g/L of aspartate family amino acids in 96 h. We also engineered a E. coli strain to produce 24 mg/L of prenol in an ethanol-containing medium, supporting the feasibility of converting ethanol into different classes of acetyl-CoA derived compounds. |
| first_indexed | 2024-09-23T07:54:38Z |
| format | Article |
| id | mit-1721.1/133403.2 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T07:54:38Z |
| publishDate | 2022 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/133403.22022-09-22T07:10:49Z Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds Liang, Hong Ma, Xiaoqiang Ning, Wenbo Liu, Yurou Sinksey, Anthony J. Stephanopoulos, Gregory Zhou, Kang Singapore-MIT Alliance in Research and Technology (SMART) Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemical Engineering © 2020 Engineering microbes to utilize non-conventional substrates could create short and efficient pathways to convert substrate into product. In this study, we designed and constructed a two-step heterologous ethanol utilization pathway (EUP) in Escherichia coli by using acetaldehyde dehydrogenase (encoded by ada) from Dickeya zeae and alcohol dehydrogenase (encoded by adh2) from Saccharomyces cerevisiae. This EUP can convert ethanol into acetyl-CoA without ATP consumption, and generate two molecules of NADH per molecule of ethanol. We optimized the expression of these two genes and found that ethanol consumption could be improved by expressing them in a specific order (ada-adh2) with a constitutive promoter (PgyrA). The engineered E. coli strain with EUP consumed approximately 8 g/L of ethanol in 96 h when it was used as sole carbon source. Subsequently, we combined EUP with the biosynthesis of polyhydroxybutyrate (PHB), a biodegradable polymer derived from acetyl-CoA. The engineered E. coli strain carrying EUP and PHB biosynthetic pathway produced 1.1 g/L of PHB from 10 g/L of ethanol and 1 g/L of aspartate family amino acids in 96 h. We also engineered a E. coli strain to produce 24 mg/L of prenol in an ethanol-containing medium, supporting the feasibility of converting ethanol into different classes of acetyl-CoA derived compounds. 2022-07-11T14:34:30Z 2021-10-27T19:52:39Z 2022-07-11T14:34:30Z 2021 2021-06-15T11:52:30Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133403.2 en 10.1016/j.ymben.2020.11.010 Metabolic Engineering Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/octet-stream Elsevier BV Other repository |
| spellingShingle | Liang, Hong Ma, Xiaoqiang Ning, Wenbo Liu, Yurou Sinksey, Anthony J. Stephanopoulos, Gregory Zhou, Kang Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title | Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title_full | Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title_fullStr | Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title_full_unstemmed | Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title_short | Constructing an ethanol utilization pathway in Escherichia coli to produce acetyl-CoA derived compounds |
| title_sort | constructing an ethanol utilization pathway in escherichia coli to produce acetyl coa derived compounds |
| url | https://hdl.handle.net/1721.1/133403.2 |
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