Current immobiliza;on techniques oWen face significant challenges, including unevendistribu;on of enzymes, reduced enzyma;c ac;vity, and material waste, all of which can limitthe effec;veness of immobilized enzymes in industrial applica;ons. To address these issues,this study introduces a hydraulic spray automa;on system specifically designed for theimmobiliza;on of redox enzymes on tex;le substrates, providing a novel, resource-efficient,and con;nuous method for enzyme immobiliza;on. For this, glucose oxidase (GOx) waschosen as the model enzyme and was immobilized onto polyester tex;les (PET) using thehydraulic spray system. To op;mize the immobiliza;on process, several cri;cal parameters,including enzyme concentra;on, pickup percenatge, and the technique of layer-by-layerassembly of GOx was studied. Results revealed significant improvements in both the stabilityand ac;vity of the immobilized GOx, which are crucial for its effec;veness in industrialapplica;ons. Results showed that, the loading of GOx on PET had no influence on type of PET(ac;vated or pris;ne) used. However, surface ac;va;on offered be2er stability of GOxagainst rinse cycles. Results also showed that, with the increase of pickup percentage theac;vity of immobilized GOx increases un;l it reaches to the plateau. The results werepromising and demonstrated significance in uniformity and reduc;on in waste and ;meduring enzyme immobiliza;on. This immobiliza;on method not only minimizes resourceconsump;on by reducing waste but also facilitates large-scale produc;on, making itpar;cularly well-suited to meet the evolving demands of the industry. Ul;mately, this researchdemonstrates the poten;al of hydraulic spray automa;on technology to revolu;onize enzymeimmobiliza;on, paving the way for more sustainable and efficient industrial applica;ons.