Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i>
Heme is of great significance in food nutrition and food coloring, and the successful launch of artificial meat has greatly improved the application of heme in meat products. The precursor of heme, 5-aminolevulinic acid (ALA), has a wide range of applications in the agricultural and medical fields,...
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/24/15524 |
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| author | Zhexian Geng Jinxia Ge Wei Cui Hui Zhou Jieying Deng Baocai Xu |
| author_facet | Zhexian Geng Jinxia Ge Wei Cui Hui Zhou Jieying Deng Baocai Xu |
| author_sort | Zhexian Geng |
| collection | DOAJ |
| description | Heme is of great significance in food nutrition and food coloring, and the successful launch of artificial meat has greatly improved the application of heme in meat products. The precursor of heme, 5-aminolevulinic acid (ALA), has a wide range of applications in the agricultural and medical fields, including in the treatment of corona virus disease 2019 (COVID-19). In this study, <i>E. coli</i> recombinants capable of heme production were developed by metabolic engineering and membrane engineering. Firstly, by optimizing the key genes of the heme synthesis pathway and the screening of hosts and plasmids, the recombinant strain EJM-pCD-AL produced 4.34 ± 0.02 mg/L heme. Then, the transport genes of heme precursors <i>CysG</i>, <i>hemX</i> and <i>CyoE</i> were knocked out, and the extracellular transport pathways of heme Dpp and Ccm were strengthened, obtaining the strain EJM-ΔCyoE-pCD-AL that produced 9.43 ± 0.03 mg/L heme. Finally, fed-batch fermentation was performed in a 3-L fermenter and reached 28.20 ± 0.77 mg/L heme and 303 ± 1.21 mg/L ALA. This study indicates that <i>E. coli</i> recombinant strains show a promising future in the field of heme and ALA production. |
| first_indexed | 2024-03-09T16:20:45Z |
| format | Article |
| id | doaj.art-79def5e03c234d20bd5e18f69ea728fc |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T16:20:45Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-79def5e03c234d20bd5e18f69ea728fc2023-11-24T15:22:58ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123241552410.3390/ijms232415524Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i>Zhexian Geng0Jinxia Ge1Wei Cui2Hui Zhou3Jieying Deng4Baocai Xu5School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, ChinaSchool of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, ChinaSchool of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, ChinaSchool of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, ChinaEngineering Research Center of Bio-Process, Ministry of Education, Hefei University of Technology, Hefei 230601, ChinaSchool of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, ChinaHeme is of great significance in food nutrition and food coloring, and the successful launch of artificial meat has greatly improved the application of heme in meat products. The precursor of heme, 5-aminolevulinic acid (ALA), has a wide range of applications in the agricultural and medical fields, including in the treatment of corona virus disease 2019 (COVID-19). In this study, <i>E. coli</i> recombinants capable of heme production were developed by metabolic engineering and membrane engineering. Firstly, by optimizing the key genes of the heme synthesis pathway and the screening of hosts and plasmids, the recombinant strain EJM-pCD-AL produced 4.34 ± 0.02 mg/L heme. Then, the transport genes of heme precursors <i>CysG</i>, <i>hemX</i> and <i>CyoE</i> were knocked out, and the extracellular transport pathways of heme Dpp and Ccm were strengthened, obtaining the strain EJM-ΔCyoE-pCD-AL that produced 9.43 ± 0.03 mg/L heme. Finally, fed-batch fermentation was performed in a 3-L fermenter and reached 28.20 ± 0.77 mg/L heme and 303 ± 1.21 mg/L ALA. This study indicates that <i>E. coli</i> recombinant strains show a promising future in the field of heme and ALA production.https://www.mdpi.com/1422-0067/23/24/155245-aminolevulinic acidheme<i>Escherichia coli</i>membrane engineeringmetabolic engineering |
| spellingShingle | Zhexian Geng Jinxia Ge Wei Cui Hui Zhou Jieying Deng Baocai Xu Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> International Journal of Molecular Sciences 5-aminolevulinic acid heme <i>Escherichia coli</i> membrane engineering metabolic engineering |
| title | Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> |
| title_full | Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> |
| title_fullStr | Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> |
| title_full_unstemmed | Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> |
| title_short | Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i> |
| title_sort | efficient de novo biosynthesis of heme by membrane engineering in i escherichia coli i |
| topic | 5-aminolevulinic acid heme <i>Escherichia coli</i> membrane engineering metabolic engineering |
| url | https://www.mdpi.com/1422-0067/23/24/15524 |
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