title: Modular optimization of heterologous pathways for de novo synthesis of (2S)-naringenin in Escherichia coli.
abstract: Due to increasing concerns about food safety and environmental issues, bio-based production of flavonoids from safe, inexpensive, and renewable substrates is increasingly attracting attention. Here, the complete biosynthetic pathway, consisting of 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase (DAHPS), chorismate mutase/prephenate dehydrogenase (CM/PDH), tyrosine ammonia lyase (TAL), 4-coumarate:CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), malonate synthetase, and malonate carrier protein, was constructed using pre-made modules to overproduce (2S)-naringenin from D-glucose. Modular pathway engineering strategies were applied to the production of the flavonoid precursor (2S)-naringenin from L-tyrosine to investigate the metabolic space for efficient conversion. Modular expression was combinatorially tuned by modifying plasmid gene copy numbers and promoter strengths to identify an optimally balanced pathway. Furthermore, a new modular pathway from D-glucose to L-tyrosine was assembled and re-optimized with the identified optimal modules to enable de novo synthesis of (2S)-naringenin. Once this metabolic balance was achieved, the optimum strain was capable of producing 100.64 mg/L (2S)-naringenin directly from D-glucose, which is the highest production titer from D-glucose in Escherichia coli. The fermentation system described here paves the way for the development of an economical process for microbial production of flavonoids.
authors.affiliation: School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Wu
authors.affiliation: School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Zhou
authors.affiliation: School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China; Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.affiliation: School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China; Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Zhou
authors.affiliation: School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China; Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D053898:Biosynthetic Pathways; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D044950:Flavanones / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D007218:Industrial Microbiology
chemical_list: D044950:Flavanones; C005273:naringenin