Abstract
Nuclease P1 (NP1) is critical for producing 5'-nucleotides, which are essential flavor enhancers in the food industry. Traditional use of free NP1 is hampered by poor reusability, high costs, and potential residual enzyme protein contamination, compromising product quality. This study introduces a novel immobilization technique using a cross-linking approach with food-grade ion exchange resin AER1 to create NP1@AER1-GA. This method achieves an enzyme activity of 51,015 U/g, with a notable immobilization yield of 67.7%. The immobilized NP1@AER1-GA exhibits significantly enhanced stability and catalytic efficiency while ensuring industrial compatibility and maintaining stringent safety standards. Under optimized conditions, NP1@AER1-GA demonstrates exceptional performance in 5'-nucleotide production, retaining approximately 85% of its initial activity after 10 cycles of reuse. This breakthrough not only boosts the efficiency and sustainability of nucleotide synthesis but also offers a scalable solution for industrial applications, promoting sustainable manufacturing practices within the food industry. By addressing key challenges associated with traditional enzymatic methods, this immobilization technique sets a new benchmark for biocatalyst design in the food processing industry.