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July 17th, 2025
Version: 2
East China University of Science and Technology
microbiology
biorxiv

Enhancing and stabilizing a high-yield industrial Rhodobacter sphaeroides strain for coenzyme Q10 production

Liang, M.Open in Google Scholar•He, X.Open in Google Scholar•Lv, D.Open in Google Scholar•Liu, J.Open in Google Scholar•Wang, K.Open in Google Scholar•Hou, Y.Open in Google Scholar•Wang, W.Open in Google Scholar•Bai, L.Open in Google Scholar•Liu, G.Open in Google Scholar•Zhu, Z.Open in Google Scholaret al.

Strengthening high-yield phenotypes while maintaining physiological and genetic stability presents a significant challenge in the improvement of high-yield industrial strains (HIS). Coenzyme Q10 (CoQ10), a crucial quinone electron carrier in the electron transport chain, is widely used in the prevention and treatment of cardiovascular diseases. In this study, the established HIS Rhodobacter sphaeroides HY01, employed for CoQ10 production, was engineered to enhance productivity while ensuring strain stability. Comparative omics identified the PrrAB two-component system as an oxygen-responsive regulator that links CoQ10 biosynthesis to photosynthetic pathways. Mutagenesis of PrrA, guided by AlphaFold3 modeling and fluorescence screening, introduced mutations that led to a 37.5% increase in CoQ10 production. To address phenotypic reversion due to metabolic burden, genome-scale CRISPR interference (CRISPRi) screening identified key genes involved in DNA repair and stress adaptation. Deletions of these genes generated a stable strain that achieved 3.6 g/L CoQ10 in a 50-L pilot-scale fed-batch fermentation, surpassing previous reports. This study reveals PrrAB-mediated flux partitioning for redox homeostasis and provides a framework for stabilizing burdened phenotypes in photosynthetic microbes, advancing the sustainable production of redox-active metabolites.

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