[P004] Poster

1:00 PM - 1:50 PM JST(4:00 AM - 4:50 AM UTC)

[P02-200] Sustainable Production through Spatial Niche Partitioning in Engineered for High-level Secretion of CO₂-derived Sucrose

○Hao Gao^1,2,3, Jee Loon Foo^1,2,3, Fengxue Xin⁴, Matthew Wook Chang^1,2,3 (1. National University of Singapore (Singapore), 2. Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore (Singapore), 3. National Centre for Engineering Biology (NCEB) (Singapore), 4. College of Biotechnology and Pharmaceutical Engineering, State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University (China))

Keywords:

Sustainability,Light-driven microbial community,Spatial niches,Manufacturing

Light-driven microbial communities (phototrophs and heterotrophs) are promising for converting CO2 into valuable chemicals. However, conventional artificial consortia often suffer from instability due to competition, where dominant strains outcompete others, leading to collapse or inefficiency. Inspired by spatial structure in natural microbial ecosystems, the hypothesis is that physical separation of community members into distinct spatial niches can reduce competition, facilitate coexistence, and enhance productivity. Herein, a synthetic light-driven community was constructed, comprising an autotrophic cyanobacterium, Synechococcus elongatus FL130, engineered for high-level sucrose secretion from CO2, and a heterotrophic yeast, Meyerozyma guilliermondii, engineered for the efficient synthesis of target chemicals. Then, we developed spatially arranged two-tiered structural living material scaffold. First, using droplet microfluidics, the individual microbial strains were encapsulated within core-shell microgels (Gel-MA@CMC), creating discrete, protected niches that prevent direct competitive exclusion while allowing for the exchange of metabolites. Second, these cell-laden microgels were formulated into a shear-thinning bioink with a secondary polymer network (SA-PBA@PVA-GMA) and assembled into robust, macroscopic living material scaffolds via extrusion-based 3D bioprinting. This process enabled the creation of a modular, scalable, and reusable biocatalytic system.
Results demonstrated a profound improvement in both community stability and productivity. The spatially partitioned system achieved production titers for 2-phenylethanol (2-PE) and tyrosol that were up to 32 and 42 times higher than those observed in conventional, unstructured liquid co-cultures. Quantitative real-time PCR (qPCR) analysis revealed that this enhancement was underpinned by a significant upregulation of key metabolic genes within the heterotrophic strain, confirming that the structured environment promoted a more efficient division of labor and metabolic cooperation. Furthermore, the macroscopic scaffold exhibited exceptional durability, maintaining high productivity over six consecutive fermentation batches without significant degradation, highlighting its potential for long-term, continuous bioprocessing.
In conclusion, this work establishes spatial niche partitioning via 3D-printed living materials as a powerful and effective strategy for engineering robust and highly efficient microbial consortia. The developed platform is not only stable and reusable but also highly modular, offering a "plug-and-play" capability to produce a variety of chemicals by simply substituting the heterotrophic strain. This innovative approach holds strong industrial potential, paving the way for the development of a truly sustainable bioeconomy built on the direct conversion of CO2 into valuable products.