Metabolic engineering of Escherichia coli W3110 strain by incorporating genome-level modifications and synthetic plasmid modules to enhance L-Dopa production from glycerol

Preparative Biochemistry & Biotechnology
Arunangshu DasKrishna Jyoti Mukherjee

Abstract

L-Tyrosine which is one of the terminal metabolites of highly regulated aromatic amino-acid biosynthesis pathway in Escherichia coli is a precursor for synthesis of L-Dopa. In this study, we report over production of L-Dopa by enhancing expression of rate limiting isoenzyme of shikimate kinase (aroL), chorismate synthase (aroC), aromatic-amino-acid aminotransferase (tyrB) and 3-phosphoshikimate 1-carboxyvinyltransferase (aroA) form a plasmid module harboring five enzymes under two inducible promoters converting shikimate to tyrosine. 4-hydroxyphenylacetate-3-hydrolase (hpaBC) which converts L-Tyrosine to L-Dopa was expressed constitutively from a separate plasmid module. Feedback deregulated expression of 3-Deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) synthase (aroG*) replacing wild type aroG under its natural promoter led to enhancement of L-Dopa production. Deletion of transcriptional repressor tyrR and links to other competing pathways improved titers of L-Dopa. We focused on having a balanced flux by constitutive expression of pathway enzymes from plasmid constructs rather than achieving higher amounts of catalytic protein by induction. We observed glycerol when used as a carbon source for the final strain led to low aci...Continue Reading

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Citations

Sep 20, 2019·Journal of Industrial Microbiology & Biotechnology·Weizhu ZengJingwen Zhou
Jul 10, 2020·Journal of Industrial Microbiology & Biotechnology·Zhu LiDawei Zhang
Jan 22, 2019·Frontiers in Bioengineering and Biotechnology·Behrouz Mohammadi NargesiJung-Won Youn
Jun 29, 2021·Applied Microbiology and Biotechnology·Akira NakagawaHiromichi Minami

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Methods Mentioned

BETA
phosphotransferase
PCR

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