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Bio-Based Production of Uroporphyrin in Escherichia coli

Synthetic Biology and Engineering. 2024, 2(1), 10002; https://doi.org/10.35534/sbe.2024.10002
Bahareh Arab    Adam W. Westbrook    Murray Moo-Young    Yilan Liu    C. Perry Chou *   
Department of Chemical Engineering, University of Waterloo, Waterloo, ON N2L 3G1, Canada
*
Authors to whom correspondence should be addressed.

Received: 14 Dec 2023    Accepted: 01 Feb 2024    Published: 06 Feb 2024   

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Abstract

Uroporphyrin (UP) is a porphyrin compound with medical applications and a key precursor for heme biosynthesis. However, there is no biosynthetic strategy for UP production. In this study, we present a novel bioprocess for enhanced production of UP in engineered Escherichia coli. We first implemented the Shemin/C4 pathway heterologously in an E. coli strain with an enlarged intracellular pool of succinyl-CoA. Using a plasmid with the trc promoter regulating the expression of a synthesized gene operon, the effects of key pathway genes, including hemA, hemB, hemC, and hemD, on UP biosynthesis were characterized. By cultivating the resulting engineered E. coli strains in a batch bioreactor with 30 g/L glycerol under aerobic conditions, up to 901.9 mg/L UP was produced. Most of the synthesized UP was extracellularly secreted with a high purity more than 80 wt%, facilitating its downstream purification. The study paves the way for large-scale bio-based production of UP using synthetic biology and metabolic engineering strategies.

Keywords

Bio-based production; Escherichia coli; Metabolic engineering; Shemin pathway; Tetrapyrrole biosynthetic pathway; Uroporphyrin

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© 2024 by the authors; licensee SCIEPublish, SCISCAN co. Ltd. This article is an open access article distributed under the CC BY license (https://creativecommons.org/licenses/by/4.0/).