The Maillard reaction between asparagine and a carbonyl source can lead to acrylamide formation, which is classified as "probably carcinogenic to humans". This study aimed to investigate the impact of varying amounts, incubation times, and temperatures of asparaginases on reducing acrylamide in wheat and rye cookies while preserving sensory attributes. To determine if different asparaginases reduce acrylamide without negative effects on cookie quality, acrylamide was quantified using ELISA, and color, texture and sensory attributes were assessed. Adding asparaginases resulted in a reduction of acrylamide by up to 85%. Incubating cookie dough for 10 or 30 min at 60 °C or 90 °C did not impact acrylamide formation. Acrylamide concentrations were higher by an average of 78 μg/kg for an incubation temperature of 90 °C compared to 60 °C. Rye cookies exhibited higher acrylamide concentrations by 490 μg/kg compared to wheat due to higher free asparagine in rye flour. Asparaginases showed minimal effects on cookie color and texture, with no changes in sensory evaluation. Acrylamide can be reduced using asparaginases, providing a simple and effective means of ensuring safe food for consumers. •Higher dough incubation temperature (90 °C vs. 60 °C) increased acrylamide•Longer dough incubation time had no effect on acrylamide formation•Increasing asparaginase concentration had little effect on acrylamide reduction•Optimal incubation temperature and time reduced acrylamide by up to 85%•Asparaginases showed little or no effect on flavor, color and texture of cookies