Coordination of GMP ligand with Cu to enhance the multiple enzymes stability and substrate specificity by co-immobilization process

Enhancing activity of multiple enzymes by co-localization are key topics in bionanotechnology. Herein, we introduce a facile co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP) in the matrix of Cu (copper) and GMP (guanosine 5′-monophosphate). The multienzyme system by co-immobilizing GOx&HRP observed with high catalytic efficiency and stability compared with free GOx&HRP enzymes. GOx&HRP@Cu/GMP stability was observed 40% increased at pH 3 and thermal stability more than 70% determined at 90 ᵒC. The storage stability was sustained 8 times higher than free enzymes after thirteen days and reusability after reacting eight times with fresh substrate retains 90% relative activity. Further, GOx&HRP@Cu/GMP demonstrated an excellent selectivity in the development of glucose biosensor and exhibited good resistance against the protease enzymes and organic solvents. The obtained results emphasize that co-immobilization of GOx&HRP in the matrix of Cu/GMP could be an efficient protocol for the biocatalyst encapsulation.