Engineering Escherichia coli for high-yield production of ectoine
Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered Escherichia coli...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co. Ltd.
2023-06-01
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| Series: | Green Chemical Engineering |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666952821000662 |
| _version_ | 1827934004887683072 |
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| author | Daoan Wang Jiamin Chen Yang Wang Guocheng Du Zhen Kang |
| author_facet | Daoan Wang Jiamin Chen Yang Wang Guocheng Du Zhen Kang |
| author_sort | Daoan Wang |
| collection | DOAJ |
| description | Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered Escherichia coli strains for the high-yield production of ectoine. The ectABC gene cluster from the native ectoine producer Halomonas elongata was introduced into different Escherichia coli (E. Coil) strains via plasmids and 0.8 g L-1 of ectoine was produced in flask cultures by engineered E. coli BL21 (DE3). Subsequently, we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes ectA, ectB, and ectC, which increased the ectoine yield to 1.6 g L-1. After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant (G1A, C932T) and the H. elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor, the titer of ectoine reached to 5.5 g L-1 in flask cultures. Finally, the engineered strain produced 60.7 g L-1 ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose. |
| first_indexed | 2024-03-13T07:32:27Z |
| format | Article |
| id | doaj.art-5c1ae8b7e31642a1be22c32fbe3c178f |
| institution | Directory Open Access Journal |
| issn | 2666-9528 |
| language | English |
| last_indexed | 2024-03-13T07:32:27Z |
| publishDate | 2023-06-01 |
| publisher | KeAi Communications Co. Ltd. |
| record_format | Article |
| series | Green Chemical Engineering |
| spelling | doaj.art-5c1ae8b7e31642a1be22c32fbe3c178f2023-06-04T04:24:32ZengKeAi Communications Co. Ltd.Green Chemical Engineering2666-95282023-06-0142217223Engineering Escherichia coli for high-yield production of ectoineDaoan Wang0Jiamin Chen1Yang Wang2Guocheng Du3Zhen Kang4The Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, ChinaThe Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, ChinaThe Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; Corresponding author.The Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, ChinaThe Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; Corresponding author.Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered Escherichia coli strains for the high-yield production of ectoine. The ectABC gene cluster from the native ectoine producer Halomonas elongata was introduced into different Escherichia coli (E. Coil) strains via plasmids and 0.8 g L-1 of ectoine was produced in flask cultures by engineered E. coli BL21 (DE3). Subsequently, we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes ectA, ectB, and ectC, which increased the ectoine yield to 1.6 g L-1. After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant (G1A, C932T) and the H. elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor, the titer of ectoine reached to 5.5 g L-1 in flask cultures. Finally, the engineered strain produced 60.7 g L-1 ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.http://www.sciencedirect.com/science/article/pii/S2666952821000662EctoineEscherichia coliMetabolic engineeringRBS design |
| spellingShingle | Daoan Wang Jiamin Chen Yang Wang Guocheng Du Zhen Kang Engineering Escherichia coli for high-yield production of ectoine Green Chemical Engineering Ectoine Escherichia coli Metabolic engineering RBS design |
| title | Engineering Escherichia coli for high-yield production of ectoine |
| title_full | Engineering Escherichia coli for high-yield production of ectoine |
| title_fullStr | Engineering Escherichia coli for high-yield production of ectoine |
| title_full_unstemmed | Engineering Escherichia coli for high-yield production of ectoine |
| title_short | Engineering Escherichia coli for high-yield production of ectoine |
| title_sort | engineering escherichia coli for high yield production of ectoine |
| topic | Ectoine Escherichia coli Metabolic engineering RBS design |
| url | http://www.sciencedirect.com/science/article/pii/S2666952821000662 |
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