SpyCatcher/SpyTag-Mediated Self-Assembly of a Supramolecular Complex for Improved Biocatalytic Production of Trehalose

Trehalose has been utilized in the food, cosmetic, and pharmaceutical industries due to its ability to stabilize biomolecules. In this study, we developed a strategy of constructing a self-assembling supramolecular multienzyme complex for the production of trehalose using SpyCatcher and SpyTag. Maltooligosyl trehalose synthase (MTS) and maltooligosyl trehalose trehalohydrolase (MTH) were fused to SpyCatcher and its corresponding ligand, SpyTag. The fused MTS-SpyCatcher (MTS-SC) and MTH-SpyTag (MTH-ST) self-assembled into a supramolecular multienzyme complex (MTS-MTH) via the isopeptide bonds formed between SpyCatcher and SpyTag. MTS-MTH showed the faster reaction rate, which reached the peak of trehalose conversion rate 10 h earlier than the free enzymes. The results demonstrate that the multienzyme self-assembly via the protein-peptide interactions of SpyCatcher and SpyTag can increase the catalytic efficiency of sequential reactions.