Efficient synthesis of l-DOPA in Escherichia coli via cofactor and enzyme engineering
Gespeichert in:
| Titel: | Efficient synthesis of l-DOPA in Escherichia coli via cofactor and enzyme engineering |
|---|---|
| Autoren: | Lihao Deng, Jurong Ping, Zhuoyuan Liu, Kai Linghu, Heng Zhang, Xiaoyu Shan, Weizhu Zeng, Jianghua Li, Jingwen Zhou |
| Quelle: | Synthetic and Systems Biotechnology, Vol 11, Iss , Pp 226-236 (2026) |
| Verlagsinformationen: | KeAi Communications Co., Ltd., 2026. |
| Publikationsjahr: | 2026 |
| Bestand: | LCC:Biotechnology LCC:Biology (General) |
| Schlagwörter: | l-DOPA, Metabolic flux, Cofactor engineering, Enzyme engineering, Enzyme substrate tunnel, Biotechnology, TP248.13-248.65, Biology (General), QH301-705.5 |
| Beschreibung: | The global incidence of Parkinson's disease continues to rise. Levodopa (l-DOPA) is the core therapeutic drug, and efficient and sustainable production methods are needed. However, the complex metabolic pathways and the low catalytic efficiency of enzymes limit biosynthesis of l-DOPA in microorganisms. To address this issue, this study significantly enhanced the production efficiency of l-DOPA through a multi-dimensional, integrated metabolic and enzyme engineering approach. Firstly, the de novo synthesis pathway for l-DOPA was established through optimization of the promoter, ribosome-binding site (RBS), plasmid copy number, and tighly accurately regulating the expression level of key enzymes. Secondly, combined with metabonomic analysis, carbon metabolic flow was diverted, increasing the l-DOPA titer by 36.7 %. Glucose dehydrogenase (BmgdH) and gluconate kinase (gntK) were introduced to construct a cofactor regeneration system, which synergistically enhanced the supply of NADH and FADH2, increasing the l-DOPA conversion rate by 18 %. Next, the substrate tunnel of 4-hydroxyphenylacetic acid-3-monooxygenase subunit B (HpaB) was subjected to rational design, and mutant T292A significantly expanded the substrate channel, improved catalytic efficiency, and decreased l-tyrosine by 87 %. Finally, through the process optimization in a 5 L bioreactor (involving phased pH control and induction timing adjustment) achieved an l-DOPA titer of 60.73 g/L, the highest reported to date for de novo microbial synthesis. This research offers a novel approach for industrial biosynthesis of l-DOPA, and broadens engineering concepts for efficient synthesis of aromatic compounds. |
| Publikationsart: | article |
| Dateibeschreibung: | electronic resource |
| Sprache: | English |
| ISSN: | 2405-805X |
| Relation: | http://www.sciencedirect.com/science/article/pii/S2405805X25001528; https://doaj.org/toc/2405-805X |
| DOI: | 10.1016/j.synbio.2025.09.011 |
| Zugangs-URL: | https://doaj.org/article/57a432668b0941d689d3f85c2ee92029 |
| Dokumentencode: | edsdoj.57a432668b0941d689d3f85c2ee92029 |
| Datenbank: | Directory of Open Access Journals |
Full Text 
Full Text Finder 
Nájsť tento článok vo Web of Science | Abstract: | The global incidence of Parkinson's disease continues to rise. Levodopa (l-DOPA) is the core therapeutic drug, and efficient and sustainable production methods are needed. However, the complex metabolic pathways and the low catalytic efficiency of enzymes limit biosynthesis of l-DOPA in microorganisms. To address this issue, this study significantly enhanced the production efficiency of l-DOPA through a multi-dimensional, integrated metabolic and enzyme engineering approach. Firstly, the de novo synthesis pathway for l-DOPA was established through optimization of the promoter, ribosome-binding site (RBS), plasmid copy number, and tighly accurately regulating the expression level of key enzymes. Secondly, combined with metabonomic analysis, carbon metabolic flow was diverted, increasing the l-DOPA titer by 36.7 %. Glucose dehydrogenase (BmgdH) and gluconate kinase (gntK) were introduced to construct a cofactor regeneration system, which synergistically enhanced the supply of NADH and FADH2, increasing the l-DOPA conversion rate by 18 %. Next, the substrate tunnel of 4-hydroxyphenylacetic acid-3-monooxygenase subunit B (HpaB) was subjected to rational design, and mutant T292A significantly expanded the substrate channel, improved catalytic efficiency, and decreased l-tyrosine by 87 %. Finally, through the process optimization in a 5 L bioreactor (involving phased pH control and induction timing adjustment) achieved an l-DOPA titer of 60.73 g/L, the highest reported to date for de novo microbial synthesis. This research offers a novel approach for industrial biosynthesis of l-DOPA, and broadens engineering concepts for efficient synthesis of aromatic compounds. |
|---|---|
| ISSN: | 2405805X |
| DOI: | 10.1016/j.synbio.2025.09.011 |