This study investigates the potential of deep eutectic solvents (DES) to enhance the biodegradation of natural rubber gloves (NRG) by Klebsiella aerogenes. Choline chloride and urea (ChCl: urea) was the DES employed to pretreat NRG at various temperatures (80 degrees C to 140 degrees C) and durations (0.5 h to 5 h). Pre-treated rubber (p-NRG) underwent significant physical and chemical changes, enhancing its biodegradability. Analytical techniques such as dry weight analysis, bacteria cell concentration, FTIR TGA, and SEM were used to characterize the pre-treated and biodegraded samples. The results have demonstrated a significant weight loss and structural modifications in p-NRG, with the highest degradation of 43 % observed at 140 degrees C for 5 hours of pretreatment before biodegradation. Meanwhile, merely 17 % of weight loss was observed when pre-treatment was not employed. DES pretreatment notably enhanced NRG biodegradability, achieving a 50.6 % weight loss when biodegradation was conducted at pH 7 and 35 degrees C. The highest cell concentration, 0.75 g/L, was recorded in the second week of the biodegradation process. Results have indicated that the maximum protein concentration of 697.3 mu g/ml, along with the highest enzyme activities for laccase and manganese peroxidase (MnP) at 0.46 f 0.05 IU and 0.30 f 0.05 IU respectively, were recorded in the second week of the biodegradation process. DES pre-treatment has significantly improved the biodegradability of NRG by Klebsiella aerogenes, offering a promising and eco-friendly solution for rubber waste management.