Sensitive, High-throughput, Metabolic Analysis by Molecular Sensors on the Membrane Surface of Mother Yeast Cells (MOMS)

Due to its genetic similarity to humans, yeast serves as a vital model organism in life sciences and medicine, allowing for the study of crucial biological processes such as cell division and metabolism for drug development. However, current tools for measuring yeast extracellular secretion lack the sensitivity, throughput, and speed required for large-scale metabolic analysis. Here, we present an ultrasensitive, large-scale analysis of yeast extracellular secretion using molecular sensors on the membrane surface of yeast cells, referred to as MOMS. We developed highly sensitive MOMS that are explicitly anchored to and confined within mother yeast cells, preventing their transfer to daughter cells during budding. MOMS enable high-throughput screening and rapid single-yeast assays. Their limit of detection (LOD) is 100 nM, and they can screen over 107 single cells per run, exceeding existing methods by more than twofold. We achieved more than a 30-fold speed boost compared to traditional droplet screens, allowing us to identify the top 0.05% of secretory strains from 2.2 × 10^6 variants in just 12 minutes. Our MOMS hold transformative potential for large-scale single-yeast metabolism analysis and bio-fabrication.