Intracellular protein ligation and polyprotein synthesis using an asparaginyl endopeptidase core

Abstract

An intracellular protein ligation system using a truncated variant of OaAEP1 (OaAEP1 core) to ligate and polymerize Ig domains in E. coli is developed, enabling real-time polyprotein synthesis without enzyme activation and external ligation. This approach yields functional, mechanically stable polyproteins and expands AEP applications in synthetic biology and biomaterials.