Metabolic engineering Escherichia coli for efficient production of icariside D2
Abstract Background Icariside D2 is a plant-derived natural glycoside with pharmacological activities of inhibiting angiotensin-converting enzyme and killing leukemia cancer cells. Production of icariside D2 by plant extraction and chemical synthesis is inefficient and environmentally unfriendly. Mi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-019-1601-x |
| _version_ | 1818230309098881024 |
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| author | Xue Liu Lingling Li Jincong Liu Jianjun Qiao Guang-Rong Zhao |
| author_facet | Xue Liu Lingling Li Jincong Liu Jianjun Qiao Guang-Rong Zhao |
| author_sort | Xue Liu |
| collection | DOAJ |
| description | Abstract Background Icariside D2 is a plant-derived natural glycoside with pharmacological activities of inhibiting angiotensin-converting enzyme and killing leukemia cancer cells. Production of icariside D2 by plant extraction and chemical synthesis is inefficient and environmentally unfriendly. Microbial cell factory offers an attractive route for economical production of icariside D2 from renewable and sustainable bioresources. Results We metabolically constructed the biosynthetic pathway of icariside D2 in engineered Escherichia coli. We screened the uridine diphosphate glycosyltransferases (UGTs) and obtained an active RrUGT3 that regio-specifically glycosylated tyrosol at phenolic position to exclusively synthesize icariside D2. We put heterologous genes in E. coli cell for the de novo biosynthesis of icariside D2. By fine-tuning promoter and copy number as well as balancing gene expression pattern to decrease metabolic burden, the BMD10 monoculture was constructed. Parallelly, for balancing pathway strength, we established the BMT23–BMD12 coculture by distributing the icariside D2 biosynthetic genes to two E. coli strains BMT23 and BMD12, responsible for biosynthesis of tyrosol from preferential xylose and icariside D2 from glucose, respectively. Under the optimal conditions in fed-batch shake-flask fermentation, the BMD10 monoculture produced 3.80 g/L of icariside D2 using glucose as sole carbon source, and the BMT23–BMD12 coculture produced 2.92 g/L of icariside D2 using glucose–xylose mixture. Conclusions We for the first time reported the engineered E. coli for the de novo efficient production of icariside D2 with gram titer. It would be potent and sustainable approach for microbial production of icariside D2 from renewable carbon sources. E. coli–E. coli coculture approach is not limited to glycoside production, but could also be applied to other bioproducts. |
| first_indexed | 2024-12-12T10:32:27Z |
| format | Article |
| id | doaj.art-31292ff27aa9420889f4e0ec42a41112 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T10:32:27Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-31292ff27aa9420889f4e0ec42a411122022-12-22T00:27:19ZengBMCBiotechnology for Biofuels1754-68342019-11-0112111210.1186/s13068-019-1601-xMetabolic engineering Escherichia coli for efficient production of icariside D2Xue Liu0Lingling Li1Jincong Liu2Jianjun Qiao3Guang-Rong Zhao4Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityFrontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityAbstract Background Icariside D2 is a plant-derived natural glycoside with pharmacological activities of inhibiting angiotensin-converting enzyme and killing leukemia cancer cells. Production of icariside D2 by plant extraction and chemical synthesis is inefficient and environmentally unfriendly. Microbial cell factory offers an attractive route for economical production of icariside D2 from renewable and sustainable bioresources. Results We metabolically constructed the biosynthetic pathway of icariside D2 in engineered Escherichia coli. We screened the uridine diphosphate glycosyltransferases (UGTs) and obtained an active RrUGT3 that regio-specifically glycosylated tyrosol at phenolic position to exclusively synthesize icariside D2. We put heterologous genes in E. coli cell for the de novo biosynthesis of icariside D2. By fine-tuning promoter and copy number as well as balancing gene expression pattern to decrease metabolic burden, the BMD10 monoculture was constructed. Parallelly, for balancing pathway strength, we established the BMT23–BMD12 coculture by distributing the icariside D2 biosynthetic genes to two E. coli strains BMT23 and BMD12, responsible for biosynthesis of tyrosol from preferential xylose and icariside D2 from glucose, respectively. Under the optimal conditions in fed-batch shake-flask fermentation, the BMD10 monoculture produced 3.80 g/L of icariside D2 using glucose as sole carbon source, and the BMT23–BMD12 coculture produced 2.92 g/L of icariside D2 using glucose–xylose mixture. Conclusions We for the first time reported the engineered E. coli for the de novo efficient production of icariside D2 with gram titer. It would be potent and sustainable approach for microbial production of icariside D2 from renewable carbon sources. E. coli–E. coli coculture approach is not limited to glycoside production, but could also be applied to other bioproducts.http://link.springer.com/article/10.1186/s13068-019-1601-xIcariside D2Escherichia coliCocultureSynthetic biologyMetabolic engineering |
| spellingShingle | Xue Liu Lingling Li Jincong Liu Jianjun Qiao Guang-Rong Zhao Metabolic engineering Escherichia coli for efficient production of icariside D2 Biotechnology for Biofuels Icariside D2 Escherichia coli Coculture Synthetic biology Metabolic engineering |
| title | Metabolic engineering Escherichia coli for efficient production of icariside D2 |
| title_full | Metabolic engineering Escherichia coli for efficient production of icariside D2 |
| title_fullStr | Metabolic engineering Escherichia coli for efficient production of icariside D2 |
| title_full_unstemmed | Metabolic engineering Escherichia coli for efficient production of icariside D2 |
| title_short | Metabolic engineering Escherichia coli for efficient production of icariside D2 |
| title_sort | metabolic engineering escherichia coli for efficient production of icariside d2 |
| topic | Icariside D2 Escherichia coli Coculture Synthetic biology Metabolic engineering |
| url | http://link.springer.com/article/10.1186/s13068-019-1601-x |
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