ArticleMicrobial cell factories2026
Multi-omics-guided metabolic engineering of Limosilactobacillus reuteri for high-level 3-hydroxypropionaldehyde production from glucose.
Article in Microbial cell factories, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundLimosilactobacillus reuteri is an important probiotic chassis for producing 3-hydroxypropionaldehyde (3-HPA), a broad-spectrum antimicrobial and biochemical. However, its production is constrained by a metabolic trilemma comprising coenzyme B₁₂ auxotrophy, redox imbalance, and carbon catabolite repression (CCR). The main bottleneck currently limiting cost-effective glucose-based production is the inability to simultaneously resolve these interconnected constraints.
resultsThrough integrated multi-omics analyses of differential carbon source performance, an unexpected regulatory mechanism was obtained. Galactose orchestrates a tripartite metabolic program in which UDP-galactose may act as a signaling molecule potentially involved in alleviating CCR; redirects carbon flux through the pentose phosphate pathway to balance ATP/NADH, and transcriptionally co-activates dhaB (glycerol dehydratase) and cobA/Q (B₁₂ biosynthesis) for coordinated holoenzyme assembly. Strikingly, this native regulatory network enables 8.51 g/L 3-HPA from galactose despite its poor support for cell growth, versus 1.67 g/L from glucose which readily supports robust growth-a 5.1-fold advantage that highlights galactose's metabolic priming efficacy. Guided by these insights, an engineered strain overexpressing glycerol dehydratase was combined with process optimization and a rationally designed four-factor targeted supplementation. Synergistic supplementation with coenzyme B₁₂, ATP, KCl, and MgCl₂ achieved 28.96 g/L 3-HPA-a 17.34-fold improvement over native glucose-based performance.
conclusionsTo our knowledge, this is the first systems-level strategy for compensating the metabolic trilemma in L. reuteri by engineering carbon source-dependent regulatory networks and cofactor dynamics in a cost-effective glucose-based system, achieving the highest 3-HPA titer reported in lactobacilli in shake-flask fermentation to date. This work establishes a scalable platform for industrial 3-HPA production and provides a systems-level framework for reprogramming central carbon metabolism in probiotic chassis to address persistent bioproduction bottlenecks.
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