Protein-based scaffold enhances efficiency of enzymatic cascades

Cell-free biocatalysis is gaining popularity as an alternative to traditional chemical catalysts due to the sustainability and selectivity of enzymes. These biological catalysts are increasingly being used in the production of valuable chemicals. Recent advancements in molecular and synthetic biology have facilitated the development of new enzymatic cascades, which are sequences of reactions where each product is subsequently transformed into the next.

To enhance the efficiency of these enzymatic cascades, researchers have focused on designing biomolecule-based scaffolds that spatially arrange multi-enzyme systems within a few nanometers. However, the precise organization of enzymes with nanometric precision poses a challenge for scaffold design.

Scientists at the Center for Cooperative Research in Biomaterials (CIC biomaGUNE) have made significant progress in this area. They have developed a nanometrically organized multi-enzyme system that utilizes engineered proteins called TRAP proteins as scaffolds for biocatalysis. This innovative approach allows for precise control of spatial distribution and physicochemical properties.

The findings of their study, recently published in the journal Nature Communications, mark a significant milestone. It represents the first instance of a protein scaffold specifically designed to organize multiple enzymes at the nanoscale and concentrate reaction intermediates around the scaffolded enzymes. While DNA scaffolds had been used for similar purposes in the past, this study demonstrates the successful application of protein-based scaffolds.

Dr. Aitziber L. Cortajarena, the scientific director of the center and an Ikerbasque Research Professor, expressed the significance of their work, stating that this research showcases a novel approach to organizing enzymes and trapping reaction intermediates using protein scaffolds, a feat previously accomplished only with DNA scaffolds.

Schematic representation of a protein scaffold formed by the TRAP1 (in light blue) and TRAP3 (in purple) proteins on which the FDH1 (in red) and AlaDH3 (in green) enzymes are assembled. Credit: CIC biomaGUNE

The researchers involved in the study, including co-author Fernando López Gallego, an Ikerbasque Research Professor, have observed that the assembled multi-enzymatic systems exhibit significantly higher productivity compared to non-assembled systems. In fact, the specific productivity of these systems can be up to five times greater. Furthermore, the team has successfully immobilized the biomolecular scaffold on solid surfaces, leading to the creation of reusable heterogeneous multifunctional biocatalysts capable of performing consecutive reaction cycles.

The outcomes of this research highlight the potential of the TRAP scaffolds as tools for spatial organization, effectively enhancing the efficiency of cell-free biosynthetic pathways. The unique characteristics of the selected protein scaffold provide nanoscale spatial control, surpassing what is achievable with conventional protein scaffolds. This approach enables the production of systems that offer unparalleled control over important parameters for optimal catalytic performance. Thus, it represents a significant step forward in the pursuit of a more sustainable socioeconomic model.

The researchers emphasize that the methodology they have developed is relatively simple and modular compared to other existing approaches. They anticipate that this technology will greatly contribute to advancing the production of more stable and efficient multi-enzyme systems.

This collaborative work demonstrates the immense potential of combining protein engineering and biocatalysis, not only for enhancing catalytic activity and enzyme stability but also for maximizing the performance of spatially organized multi-enzyme systems. The applications of this approach in biocatalysis extend beyond the scope of this study and could find utility in various fields of applied science, such as integration into energy devices or the development of biocatalytic materials for industrial processes.

Source: CIC biomaGUNE

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