Implementation of activity assays in microfluidics at high temperature for esterase/lipase screening

The aim of this work is to describe the development and optimization of a microfluidic-based screening procedure for the detection of metagenomic esterases and lipases from thermal environments. The proposed approach combines droplet microfluidics with fluorogenic substrates to enable high-throughput and sensitive identification of enzymatic activity at elevated temperatures. Key parameters affecting droplet generation, assay stability, and signal detection were systematically evaluated and optimized. This procedure provides a robust platform for the functional screening of thermophilic metagenomic libraries and represents a valuable tool for the discovery of novel biocatalysts with potential biotechnological applications.