Core Viewpoint - The article discusses the successful development of a high-throughput screening system based on Surface-Enhanced Raman Spectroscopy (SERS) by Professor Wang Dan's team at Chongqing University, which allows for real-time, label-free analysis of bacterial metabolites within microdroplets, providing an efficient and sustainable technology for green biomanufacturing [2][4]. Group 1: High-Throughput Screening Platform - The RH-RMSS platform utilizes SERS technology to directly detect metabolite concentrations without the need for cell staining or labeling, processing 180 microdroplets per second, significantly faster than traditional methods [4]. - The platform successfully screened high-yield strains for various organic acids, achieving yields of 1.61 times, 1.52 times, and 1.81 times that of the original strains for succinic acid (SA), D-lactic acid (D-PLA), and acrylic acid (AA), respectively [4]. - The system reveals key enzyme mutation mechanisms and implements multi-level metabolic optimization, achieving a maximum succinic acid yield of 138.92 g/L using optimized strains [4]. Group 2: System Validation and Performance - The RH-RMSS system demonstrated high accuracy (>98%) and efficiency in screening, effectively enriching high-yield strains from a large pool of mutants [10]. - The system's ability to accurately quantify organic acids within individual microdroplets was validated, establishing its reliability as a high-throughput screening tool [8]. - The research successfully transitioned from microdroplet screening to large-scale fermentation verification, showcasing the engineered strains' capability to utilize non-food biomass as a carbon source for succinic acid production [15]. Group 3: Mechanistic Insights and Optimization Strategies - The D210G mutation was shown to enhance the catalytic efficiency of the phosphoenolpyruvate carboxylase (PEPC) enzyme, which is crucial for achieving high succinic acid yields [11]. - The study highlights the synergistic effects of neutralizing agent optimization, RBS sequence optimization, and adaptive evolution on improving succinic acid production [14]. - The RH-RMSS platform is presented as a versatile tool adaptable for screening high-yield strains for various organic acids and other metabolites with characteristic Raman signals [18].