Production of Immobilized Arginine Deiminase Using the Self-Assembling Peptide ELK16 for Efficient l-Citrulline Synthesis.

Self-assembling peptide (SAP) tags induce protein self-assembly, forming insoluble protein aggregates. Traditional l-citrulline production using arginine deiminase (ADI) is limited by enzyme instability and low reusability. SAP tags were fused with ADI to overcome these challenges, and ADI-ELK16 demonstrated optimal activity at 55 °C and pH 6.0 with enhanced thermal stability. ADI-ELK16 retained 57.34% of its enzyme activity after 10 cycles, with notable reusability. The protein was characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS), and the zeta potential. Additionally, the conversion of 100 g/L l-arginine to 92.3 g/L l-citrulline over batch reactions validated the industrial potential of ADI-ELK16. Compared with traditional immobilization methods, this approach eliminates the need for carrier materials, simplifying the immobilization process and significantly enhancing the catalytic performance and stability, making ADI-ELK16 a highly efficient and reusable system for industrial applications.