Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli
The functional reconstruction of acetoin and meso-2,3-butanediol (meso-2,3-BD) biosynthetic pathways in Escherichia coli have been explored systematically. Pathway construction involved the in vivo screening of prospective pathway isozymes of yeast and bacterial origin. After substantial engineerin...
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| Format: | Article |
| Language: | en_US |
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John Wiley & Sons, Inc.
2012
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| Online Access: | http://hdl.handle.net/1721.1/68702 https://orcid.org/0000-0003-0437-3157 |
| _version_ | 1811087768955125760 |
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| author | Nielsen, David R. Yoon, Sang-Hwal Yuan, Clara J. Prather, Kristala L. Jones |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Nielsen, David R. Yoon, Sang-Hwal Yuan, Clara J. Prather, Kristala L. Jones |
| author_sort | Nielsen, David R. |
| collection | MIT |
| description | The functional reconstruction of acetoin and meso-2,3-butanediol (meso-2,3-BD) biosynthetic
pathways in Escherichia coli have been explored systematically. Pathway construction involved the in vivo screening of prospective pathway isozymes of yeast and bacterial origin. After substantial engineering of the host background to increase pyruvate availability, E. coli YYC202(DE3) ldhA− ilvC− expressing ilvBN from E. coli and aldB from L. lactis (encoding acetolactate synthase and acetolactate decarboxylase activities, respectively) was able to produce up to 870 mg/L acetoin, with no coproduction of diacetyl observed. These strains were also found to produce small quantities of meso-2,3-BD, suggesting the existence of endogenous 2,3-BD dehydrogenase activity. Finally, the coexpression of bdh1 from S. cerevisiae, encoding 2,3-BD dehydrogenase, in this strain resulted in the production of up to 1120 mg/L meso-2,3-BD, with glucose a yield of 0.29 g/g. While disruption of the native lactate biosynthesis pathway increased pyruvate precursor availability to this strain, increased availability of NADH for acetoin reduction to meso-2,3-BD was found to be the most important consequence of ldhA deletion. |
| first_indexed | 2024-09-23T13:51:40Z |
| format | Article |
| id | mit-1721.1/68702 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:51:40Z |
| publishDate | 2012 |
| publisher | John Wiley & Sons, Inc. |
| record_format | dspace |
| spelling | mit-1721.1/687022022-09-28T16:39:02Z Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli Metabolic engineering of acetoin and meso-2, 3-butanediol biosynthesis in E.coli Nielsen, David R. Yoon, Sang-Hwal Yuan, Clara J. Prather, Kristala L. Jones Massachusetts Institute of Technology. Department of Chemical Engineering Prather, Kristala L. Jones Nielsen, David R. Yoon, Sang-Hwal Yuan, Clara J. Prather, Kristala L. Jones The functional reconstruction of acetoin and meso-2,3-butanediol (meso-2,3-BD) biosynthetic pathways in Escherichia coli have been explored systematically. Pathway construction involved the in vivo screening of prospective pathway isozymes of yeast and bacterial origin. After substantial engineering of the host background to increase pyruvate availability, E. coli YYC202(DE3) ldhA− ilvC− expressing ilvBN from E. coli and aldB from L. lactis (encoding acetolactate synthase and acetolactate decarboxylase activities, respectively) was able to produce up to 870 mg/L acetoin, with no coproduction of diacetyl observed. These strains were also found to produce small quantities of meso-2,3-BD, suggesting the existence of endogenous 2,3-BD dehydrogenase activity. Finally, the coexpression of bdh1 from S. cerevisiae, encoding 2,3-BD dehydrogenase, in this strain resulted in the production of up to 1120 mg/L meso-2,3-BD, with glucose a yield of 0.29 g/g. While disruption of the native lactate biosynthesis pathway increased pyruvate precursor availability to this strain, increased availability of NADH for acetoin reduction to meso-2,3-BD was found to be the most important consequence of ldhA deletion. Natural Sciences and Engineering Research Council of Canada Korea Research Foundation (Grant) 2012-01-30T15:33:22Z 2012-01-30T15:33:22Z 2010-01 2009-12 Article http://purl.org/eprint/type/JournalArticle 1860-6768 1860-7314 http://hdl.handle.net/1721.1/68702 Nielsen, David R. et al. “Metabolic engineering of acetoin and meso-2, 3-butanediol biosynthesis in E. coli.” Biotechnology Journal 5.3 (2010): 274-284. https://orcid.org/0000-0003-0437-3157 en_US http://dx.doi.org/10.1002/biot.200900279 Biotechnology Journal Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf John Wiley & Sons, Inc. Prof. Prather via Erja Kajosalo |
| spellingShingle | Nielsen, David R. Yoon, Sang-Hwal Yuan, Clara J. Prather, Kristala L. Jones Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title | Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title_full | Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title_fullStr | Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title_full_unstemmed | Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title_short | Engineering Acetoin and meso-2,3-Butanediol Biosynthesis in E.coli |
| title_sort | engineering acetoin and meso 2 3 butanediol biosynthesis in e coli |
| url | http://hdl.handle.net/1721.1/68702 https://orcid.org/0000-0003-0437-3157 |
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