Nagoya University Researchers Develop SMART Platform to Accelerate Custom Enzyme Engineering for Industrial Use

Nagoya University researchers unveil the SMART platform, cutting enzyme screening from weeks to days using single-molecule mRNA display technology.

By: AXL Media

Published: Apr 30, 2026, 5:00 AM EDT

Source: Information for this report was sourced from EurekAlert!

Overcoming the Bottleneck of Directed Evolution

The field of protein engineering has long relied on directed evolution to optimize natural enzymes for industrial applications, a process that traditionally mimics natural selection over several weeks. However, the introduction of artificial mutations can create a staggering 100 trillion candidate variants, making the screening process both labor-intensive and prohibitively expensive. To solve this, researchers at Nagoya University developed the SMART platform, which stands for Single-Molecule Assay on Ribonucleic acid by Translated product. According to Professor Hideo Nakano, this system represents the most efficient method for enzyme screening currently possible, as it operates with precision at the single-molecule level.

Establishing a Chemical Bridge Between Protein and Gene

The primary innovation of the SMART system lies in its ability to solve the "display" problem, where proteins and their encoding genes are typically separate. By utilizing puromycin as a chemical bridge, the researchers successfully linked each enzyme protein directly to its corresponding messenger RNA blueprint. This mRNA display technique allows scientists to track the relationship between an individual protein's performance and its genetic identity with absolute precision. This physical connection ensures that when a superior variant is discovered, its genetic code is immediately available for sequencing and further development.

A Novel Auxiliary Unit for Activity Detection

To identify which enzymes in a massive library are actually performing the desired chemical reactions, the SMART system incorporates an auxiliary detection unit. In this study, the team used an engineered version of ascorbate peroxidase 2, known as APEX2, to screen for oxidases. When a target enzyme shows activity by releasing hydrogen peroxide, the APEX2 auxiliary unit attaches a biotin marker to the molecule. This marker acts as a molecular handle, allowing the active, high-performing enzymes to be easily isolated and captured using magnetic beads for further analysis.