Abstract
Genome engineering has become a powerful tool for creating useful strains in research and industry. In this study, we applied singleplex and multiplex genome engineering approaches to construct an E. coli strain for the production of L-DOPA from glucose. We first used the singleplex genome engineering approach to create an L-DOPA-producing strain, E. coli DOPA-1, by deleting transcriptional regulators (tyrosine repressor tyrR and carbon storage regulator A csrA), altering glucose transport from the phosphotransferase system (PTS) to ATP-dependent uptake and the phosphorylation system overexpressing galactose permease gene (galP) and glucokinase gene (glk), knocking out glucose-6-phosphate dehydrogenase gene (zwf) and prephenate dehydratase and its leader peptide genes (pheLA) and integrating the fusion protein chimera of the downstream pathway of chorismate. Then, multiplex automated genome engineering (MAGE) based on 23 targets was used to further improve L-DOPA production. The resulting strain, E. coli DOPA-30N, produced 8.67 g/L of L-DOPA in 60 h in a 5 L fed-batch fermentation. This titer is the highest achieved in metabolically engineered E. coli having PHAH activity from glucose.
Highlights
Genome engineering has become a powerful tool for creating useful strains in research and industry
E. coli W hpaBC has been successfully introduced into E. coli to produce L-DOPA from glucose[16]
Tyrosine repressor (TyrR) is a transcriptional dual regulator that represses the transcription of several genes encoding enzymes involved in aromatic acid biosynthesis[23]
Summary
Genome engineering has become a powerful tool for creating useful strains in research and industry. Muñoz et al reported an engineered E. coli having PHAH activity, which can produce 1.5 g/L of L-DOPA from glucose[16]. The titer of L-DOPA in the engineered E. coli is lower than that of the microbial fermentation from tyrosine or catechol/ pyruvate.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
