This study presents the potential of digital valve-jet printing technology to immobilize enzymes on Polyester fabric. In this study, Glucose oxidase (GOx) enzyme was immobilized on surface activated non-woven polyester (PET) fabric by using this novel printing technology. The enzymatic activity and stability of the immobilized GOx was evaluated systematically and efforts were made to enhance the stability of the immobilized enzymes. For this, GOx enzymatic activity on ink for digital valve-jet printing was studied under various parameters such as viscosity, pH, surface tension, and particle size. The inert surface of PET was activated by alkaline hydrolysis. Chitosan grating was done in addition to further enhance the surface functionality of the activated PET. Furthermore, the machine parameters (number of print passes, printing pressure) of the digital valve-jet printer were also studied to investigate their effect on the activity of the immobilized GOx. Enzymatic activity was evaluated by colorimetric assay test. Viscosity of the ink was measured by using Modular compact rheometer. PET surface activation was evaluated by using Theta optical tensiometer and FTIR anal-ysis. The results showed that GOx enzymes can be successfully immobilized on surface activated PET fabric by using digital valve-jet printing. The viscosity of ink has an effect on GOx enzyme activity and ink viscosity of 3,2 mPa.s (30% glycerol) showed the best result. Results from printing machine parameter showed 1,5 bar print pressure, and one print pass as optimized parameter for GOx printing. Comparative results revealed that chitosan grafting enhanced GOx enzymatic activity and stability. GOx immobilized chitosan-grafted-alkali-activated PET exhibited 33,26% higher enzymatic activity than that of GOx immobilized alkali activated PET. Furthermore, it showed improved wash durability and retained 67,66% enzymatic activity after five rinse cycles. In addition, it showed superior storage stability, with over 86% activity retention after four weeks at 4 °C cold storage. This study demonstrated digital valve-jet printing as a promising technology for GOx enzyme immobilization on surface activated PET. These findings underscore methods to improvise GOx enzyme immobilization, operational durability, and long-term storage stability. It also represents a step toward resource efficient process development by reducing environmental footprint paving the way for the future of functional textiles.