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. 2021 Oct 19;12:6085. doi: 10.1038/s41467-021-26361-1

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Schematic illustration of the biosynthetic pathways leading to the production of DEIN and related byproducts. To ensure efficient screening of biosynthetic enzymes for DEIN production, a p-HCA producing strain QL11, harboring overexpression of enzymes responsible for both endogenous (yellow arrows) and exogenous (blue arrows) reactions, was selected as the starting strain. ARO3, DAHP synthase; ARO4*, L-tyrosine-feedback-insensitive DAHP synthase (ARO4^K229L); ARO1, pentafunctional aromatic protein; EcaroL, shikimate kinase from E. coli; ARO2, chorismate synthase; ARO7*, L-tyrosine-feedback-insensitive chorismate mutase (ARO7^G141S); AtATR2, cytochrome P450 reductase from A. thaliana; CYB5, yeast native cytochrome b5. DAHP, 3-deoxy-D-arabino-2-heptulosonic acid 7-phosphate; SHIK, shikimate; S3P, shikimate-3-phosphate; EPSP, 5-enolpyruvyl-shikimate-3-phosphate; CHA, chorismic acid; PPA, prephenate. See Fig. 1 and its legend regarding abbreviations of metabolites and other gene details. b, c Production profiles of intermediates LIG and ISOLIG produced by yeast strains harboring different combinations of biosynthetic enzymes 4CL, CHS, CHR, and CHI. d DEIN production of C09 strain overexpressing different biosynthetic genes encoding 2-HIS and HID and relevant genetic characteristics of the resultant strains. For the source of selected plant genes: Mt, Medicago truncatula; Tp, Trifolium pretense. See Fig. 1 legend regarding abbreviations of other plant species. Cells were grown in a defined minimal medium with 30 g L⁻¹ glucose as the sole carbon source, and cultures were sampled after 72 h of growth for metabolite detection. All data represent the mean of n = 3 biologically independent samples and error bars show standard deviation. The source data underlying figures (b−d) are provided in a Source Data file.