# Fabrication of a 3D nano-bienzyme cascade reactor based on a Dps protein scaffold for chiral amine synthesis

## TL;DR

Researchers built a 3D nano-enzyme reactor using a Dps protein scaffold, which efficiently and sustainably produces chiral amines with high yield and selectivity.

## Contribution

A novel 3D nano-bienzyme cascade reactor was fabricated using a Dps protein scaffold for efficient chiral amine synthesis.

## Key findings

- The 3DNECR showed significantly higher catalytic efficiency due to optimized spatial organization.
- The reactor maintained over 80% activity after 9 days and high stability across pH and temperature ranges.
- The 3DNECR achieved 99.9% yield and enantioselectivity in synthesizing R-BPA with 92% activity after six reuse cycles.

## Abstract

Based on the self-assembling properties of the SpyCatcher/SpyTag system and the structural advantages of Dps protein, this study successfully constructed a three-dimensional nano-enzyme cascade reactor (3DNECR) through the covalent self-assembly of SpyTag-ADH and SpyCatcher-Dps-ATA117 fusion proteins. The 3DNECR exhibited significantly enhanced catalytic efficiency compared to the two-dimensional control, attributed to optimized spatial organization promoting substrate channeling. The reactor exhibited remarkable storage, pH, and thermal stability. It maintained over 80% activity after 9 days of storage, showed superior pH tolerance across pH 8–10, and remained stable in the temperature range of 4–40 ℃. Molecular docking confirmed strong interfacial binding (− 17.3 kcal/mol) between assembly components and favorable substrate binding (− 7.4 kcal/mol) within the active site. Furthermore, the 3DNECR was applied to the asymmetric synthesis of (R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine (R-BPA) in an oil-water biphasic system. Under optimized conditions, the 3DNECR consistently achieved high yields (99.9%) and excellent enantioselectivity (99.9%). The 3DNECR maintained a relative enzyme activity as high as 92% even after six cycles of reuse. This integrated platform showcases substantial potential for efficient and sustainable biocatalytic applications.

The online version contains supplementary material available at 10.1186/s40643-026-01022-7.