We are pleased to highlight the publication Fluorogenic, Subsingle-Turnover Monitoring of Enzymatic Reactions Involving NAD(P)H Provides a Generalised Platform for Directed Ultrahigh-Throughput Evolution of Biocatalysts in Microdroplets. This work supports faster and more reliable enzyme screening. It addresses a key challenge in enzyme engineering for a sustainable bioeconomy.
The study introduces a flexible coupled assay. It links enzyme activity to a fluorescent signal via NAD(P)H. This removes the need for complex model substrates. It allows researchers to screen a wide range of enzymes, including challenging targets such as glycosyl hydrolases.
The method shows high sensitivity in both plate-based and microdroplet formats. It achieves low nanomolar detection and improves sensitivity by three orders of magnitude compared to absorbance systems. The signal remains stable, which ensures reliable high-throughput screening.
The platform was validated through directed evolution. A glycosidase was improved using a natural substrate from a large variant library. This demonstrates its practical value. It enables more efficient discovery and optimisation of biocatalysts.
For Bluetools, this work reflects the project’s commitment to advancing tools for the blue bioeconomy. Improved screening technologies are essential for unlocking the potential of marine microbiomes and sustainable biocatalysis.
Read the full article here.