Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate)
Acetate is a promising economical and sustainable carbon source for bioproduction, but it is also a known cell-growth inhibitor. In this study, adaptive laboratory evolution (ALE) with acetate as selective pressure was applied to <i>Halomonas bluephagenesis</i> TD1.0, a fast-growing and...
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MDPI AG
2022-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/9/3022 |
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| author | Jing Zhang Biao Jin Jing Fu Zhiwen Wang Tao Chen |
| author_facet | Jing Zhang Biao Jin Jing Fu Zhiwen Wang Tao Chen |
| author_sort | Jing Zhang |
| collection | DOAJ |
| description | Acetate is a promising economical and sustainable carbon source for bioproduction, but it is also a known cell-growth inhibitor. In this study, adaptive laboratory evolution (ALE) with acetate as selective pressure was applied to <i>Halomonas bluephagenesis</i> TD1.0, a fast-growing and contamination-resistant halophilic bacterium that naturally accumulates poly(3-hydroxybutyrate) (PHB). After 71 transfers, the evolved strain, B71, was isolated, which not only showed better fitness (in terms of tolerance and utilization rate) to high concentrations of acetate but also produced a higher PHB titer compared with the parental strain TD1.0. Subsequently, overexpression of acetyl-CoA synthetase (ACS) in B71 resulted in a further increase in acetate utilization but a decrease in PHB production. Through whole-genome resequencing, it was speculated that genetic mutations (single-nucleotide variation (SNV) in <i>phaB</i>, <i>mdh,</i> and the upstream of <i>OmpA,</i> and insertion of <i>TolA</i>) in B71 might contribute to its improved acetate adaptability and PHB production. Finally, in a 5 L bioreactor with intermittent feeding of acetic acid, B71 was able to produce 49.79 g/L PHB and 70.01 g/L dry cell mass, which were 147.2% and 82.32% higher than those of TD1.0, respectively. These results highlight that ALE provides a reliable method to harness <i>H. bluephagenesis</i> to metabolize acetate for the production of PHB or other high-value chemicals more efficiently. |
| first_indexed | 2024-03-10T03:52:40Z |
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| id | doaj.art-63a420b77058473082c938322cb4e597 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T03:52:40Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj.art-63a420b77058473082c938322cb4e5972023-11-23T08:53:02ZengMDPI AGMolecules1420-30492022-05-01279302210.3390/molecules27093022Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate)Jing Zhang0Biao Jin1Jing Fu2Zhiwen Wang3Tao Chen4Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072, ChinaFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072, ChinaDepartment of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, SwedenFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072, ChinaFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072, ChinaAcetate is a promising economical and sustainable carbon source for bioproduction, but it is also a known cell-growth inhibitor. In this study, adaptive laboratory evolution (ALE) with acetate as selective pressure was applied to <i>Halomonas bluephagenesis</i> TD1.0, a fast-growing and contamination-resistant halophilic bacterium that naturally accumulates poly(3-hydroxybutyrate) (PHB). After 71 transfers, the evolved strain, B71, was isolated, which not only showed better fitness (in terms of tolerance and utilization rate) to high concentrations of acetate but also produced a higher PHB titer compared with the parental strain TD1.0. Subsequently, overexpression of acetyl-CoA synthetase (ACS) in B71 resulted in a further increase in acetate utilization but a decrease in PHB production. Through whole-genome resequencing, it was speculated that genetic mutations (single-nucleotide variation (SNV) in <i>phaB</i>, <i>mdh,</i> and the upstream of <i>OmpA,</i> and insertion of <i>TolA</i>) in B71 might contribute to its improved acetate adaptability and PHB production. Finally, in a 5 L bioreactor with intermittent feeding of acetic acid, B71 was able to produce 49.79 g/L PHB and 70.01 g/L dry cell mass, which were 147.2% and 82.32% higher than those of TD1.0, respectively. These results highlight that ALE provides a reliable method to harness <i>H. bluephagenesis</i> to metabolize acetate for the production of PHB or other high-value chemicals more efficiently.https://www.mdpi.com/1420-3049/27/9/3022acetate<i>Halomonas bluephagenesis</i>adaptive laboratory evolutionPHB |
| spellingShingle | Jing Zhang Biao Jin Jing Fu Zhiwen Wang Tao Chen Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) Molecules acetate <i>Halomonas bluephagenesis</i> adaptive laboratory evolution PHB |
| title | Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) |
| title_full | Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) |
| title_fullStr | Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) |
| title_full_unstemmed | Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) |
| title_short | Adaptive Laboratory Evolution of <em>Halomonas bluephagenesis</em> Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate) |
| title_sort | adaptive laboratory evolution of em halomonas bluephagenesis em enhances acetate tolerance and utilization to produce poly 3 hydroxybutyrate |
| topic | acetate <i>Halomonas bluephagenesis</i> adaptive laboratory evolution PHB |
| url | https://www.mdpi.com/1420-3049/27/9/3022 |
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