[1S05e] 13. Microbial Engineering for Natural Products Synthesis - Part 3

5:30 PM - 5:45 PM JST(8:30 AM - 8:45 AM UTC)

[1MENP-16] Carbon Flux Rewiring and Light-Mediated Control of Lipstatin Biosynthesis in Streptomyces toxytricini

○Kashyap Kumar Dubey¹ (1. School of Biotechnology, Jawaharlal Nehru University (India))

Keywords:

Streptomyces,Photoreceptor,Lipstatin,Anti-Obesity Drug,Carbon-flux

Streptomyces are versatile cell factories for the production of therapeutically important secondary metabolites; however, their complex life cycle and tightly regulated metabolism often limit production efficiency. In this study, the production of lipstatin, an anti-obesity prodrug, was enhanced by addressing limitations in precursor availability, carbon flux distribution, and regulatory control. Genomic and systems-level analyses identified key metabolic pathways and biosynthetic clusters associated with lipstatin production, particularly the link between central carbon metabolism and fatty acid biosynthesis. Targeted metabolic interventions, including optimization of nutrient composition and precursor supplementation, increased the availability of branched-chain amino acids and fatty acid intermediates, thereby enhancing metabolic channeling toward lipstatin biosynthesis. To further improve production efficiency, physiological parameters and growth conditions were optimized, and iterative strain-improvement approaches yielded metabolically robust strains with enhanced biosynthetic capacity. Proteomic and metabolomic analyses highlighted the importance of coordinated regulation between growth, morphology, and secondary metabolism, emphasizing the need for dynamic control strategies. A carbon-conserving metabolic engineering approach based on the non-oxidative glycolysis (NOG) pathway was implemented to reduce carbon loss and enhance precursor generation. This strategy improved carbon utilization efficiency and contributed to increased lipstatin yield by strengthening flux toward biosynthetic intermediates. In a separate approach, light-mediated regulation was explored to control lipstatin biosynthesis. Distinct variations in biomass accumulation were observed, with red light resulting in a fourfold increase (4.2 g/L) compared to dark conditions (1 g/L). Red-light exposure also led to significantly smaller pellet formation (~81.4 ± 13 µm), approximately ninefold smaller than those formed in the dark, and was associated with the highest lipstatin titre (~6 g/L). Proteome-wide analysis using pBLAST and conserved domain searches identified two proteins containing canonical bacteriophytochrome domains, suggesting the involvement of red- and far-red-light-sensing photoreceptors in regulating downstream metabolic processes. Overall, this study demonstrates that combining carbon-conserving metabolic engineering with light-mediated regulatory control provides an effective framework for enhancing lipstatin production in Streptomyces toxytricini, offering a promising strategy for developing efficient microbial production systems for high-value secondary metabolites.