Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin
Abstract Several chromosomally expressed AceE variants were constructed in Escherichia coli ΔldhA ΔpoxB ΔppsA and compared using glucose as the sole carbon source. These variants were examined in shake flask cultures for growth rate, pyruvate accumulation, and acetoin production via heterologous exp...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2023-03-01
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| Series: | Engineering in Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/elsc.202200054 |
| _version_ | 1811161469063004160 |
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| author | W. Chris Moxley Rachel E. Brown Mark A. Eiteman |
| author_facet | W. Chris Moxley Rachel E. Brown Mark A. Eiteman |
| author_sort | W. Chris Moxley |
| collection | DOAJ |
| description | Abstract Several chromosomally expressed AceE variants were constructed in Escherichia coli ΔldhA ΔpoxB ΔppsA and compared using glucose as the sole carbon source. These variants were examined in shake flask cultures for growth rate, pyruvate accumulation, and acetoin production via heterologous expression of the budA and budB genes from Enterobacter cloacae ssp. dissolvens. The best acetoin‐producing strains were subsequently studied in controlled batch culture at the one‐liter scale. PDH variant strains attained up to four‐fold greater acetoin than the strain expressing the wild‐type PDH. In a repeated batch process, the H106V PDH variant strain attained over 43 g/L of pyruvate‐derived products, acetoin (38.5 g/L) and 2R,3R‐butanediol (5.0 g/L), corresponding to an effective concentration of 59 g/L considering the dilution. The acetoin yield from glucose was 0.29 g/g with a volumetric productivity of 0.9 g/L·h (0.34 g/g and 1.0 g/L·h total products). The results demonstrate a new tool in pathway engineering, the modification of a key metabolic enzyme to improve the formation of a product via a kinetically slow, introduced pathway. Direct modification of the pathway enzyme offers an alternative to promoter engineering in cases where the promoter is involved in a complex regulatory network. |
| first_indexed | 2024-04-10T06:14:40Z |
| format | Article |
| id | doaj.art-0d69eb95dcfc4230bef7487265ccdad7 |
| institution | Directory Open Access Journal |
| issn | 1618-0240 1618-2863 |
| language | English |
| last_indexed | 2024-04-10T06:14:40Z |
| publishDate | 2023-03-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj.art-0d69eb95dcfc4230bef7487265ccdad72023-03-02T08:33:11ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632023-03-01233n/an/a10.1002/elsc.202200054Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoinW. Chris Moxley0Rachel E. Brown1Mark A. Eiteman2Department of Microbiology University of Georgia Athens Georgia USASchool of Chemical Materials and Biomedical Engineering University of Georgia Athens Georgia USADepartment of Microbiology University of Georgia Athens Georgia USAAbstract Several chromosomally expressed AceE variants were constructed in Escherichia coli ΔldhA ΔpoxB ΔppsA and compared using glucose as the sole carbon source. These variants were examined in shake flask cultures for growth rate, pyruvate accumulation, and acetoin production via heterologous expression of the budA and budB genes from Enterobacter cloacae ssp. dissolvens. The best acetoin‐producing strains were subsequently studied in controlled batch culture at the one‐liter scale. PDH variant strains attained up to four‐fold greater acetoin than the strain expressing the wild‐type PDH. In a repeated batch process, the H106V PDH variant strain attained over 43 g/L of pyruvate‐derived products, acetoin (38.5 g/L) and 2R,3R‐butanediol (5.0 g/L), corresponding to an effective concentration of 59 g/L considering the dilution. The acetoin yield from glucose was 0.29 g/g with a volumetric productivity of 0.9 g/L·h (0.34 g/g and 1.0 g/L·h total products). The results demonstrate a new tool in pathway engineering, the modification of a key metabolic enzyme to improve the formation of a product via a kinetically slow, introduced pathway. Direct modification of the pathway enzyme offers an alternative to promoter engineering in cases where the promoter is involved in a complex regulatory network.https://doi.org/10.1002/elsc.202200054acetoinacetolactate synthasepoint mutationpyruvate dehydrogenaserepeated batch fermentation |
| spellingShingle | W. Chris Moxley Rachel E. Brown Mark A. Eiteman Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin Engineering in Life Sciences acetoin acetolactate synthase point mutation pyruvate dehydrogenase repeated batch fermentation |
| title | Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin |
| title_full | Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin |
| title_fullStr | Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin |
| title_full_unstemmed | Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin |
| title_short | Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate‐derived acetoin |
| title_sort | escherichia coli acee variants coding pyruvate dehydrogenase improve the generation of pyruvate derived acetoin |
| topic | acetoin acetolactate synthase point mutation pyruvate dehydrogenase repeated batch fermentation |
| url | https://doi.org/10.1002/elsc.202200054 |
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