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August 27th, 2025
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Northeast Forestry University
synthetic biology
bioRxiv

Efficient biosynthesis of 3-hydroxypropionic acid in recombinant Escherichia coli by metabolic engineering

Wang, Y.Open in Google Scholar•Suo, C.Open in Google Scholar•Yang, J.Open in Google Scholar•Cui, Y.Open in Google Scholar•Abdallah, M. M.Open in Google Scholar•Yang, H.Open in Google Scholar•Wang, P.Open in Google Scholar•Li, L.Open in Google Scholar•Liu, C.Open in Google Scholar

3-Hydroxypropionic acid (3-HP) is a platform compound that can produce many chemical commodities. This study focuses on establishing and optimizing the production of 3-HP in E. coli.To increase the metabolic flux of acetyl-CoA, a precursor for the synthesis of 3-HP, CRISPR/Cas9-based DNA editing technique was used to knock out the genes encoding pyruvate oxidase (poxB), lactate dehydrogenase (ldhA) and phosphate transacetylase (pta), thereby reducing the formation of by-products. Concurrently, the acetyl coenzyme a carboxylase gene (accDABC) is overexpressed on the chromosome with the objective of augmenting intracellular acetyl-CoA levels and, consequently, 3-HP production. Next, we introduced a plasmid containing a codon-optimized malonyl-CoA reductase gene (mcr) into the engineered strain. Finally, we constructed a transcription factor-based metabolite biosensor utilizing the PpHpdR/PhpdH system, followed by the screening of mutant strains for enhanced 3-HP production through adaptive laboratory evolution. Combining the above metabolic engineering efforts with optimisation of media and fermentation conditions, the 3-HP titer of the engineered strain WY7 increased from an initial titer 0.34 g/L to 48.8 g/L.

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