The carcinogen ethyl carbamate (EC) in food is a potential threat to health. Available urethanases cannot efficiently degrade EC because of their instability or low activity under acidic conditions. Here, a novel thermostable urethanase was identified in Thermoflavimicrobium dichotomicum using a database-mining approach. The enzyme displayed exceptional thermotolerance, with an optimum temperature of 75 °C, and exhibited 58.6 % of its maximum activity at 90 °C. After incubation at temperatures below 70 °C for 30 min, 100 % activity was maintained. Following treatment at 4 °C for 6 h, it retained 59-87 % of its activity at pH 4.0-5.0, demonstrating the highest acid stability reported so far. This enzyme showed good ethanol tolerance. 80.4 % of its activity was retained after incubation in 10 % (v/v) ethanol solution at 37 °C for 1 h. The enzyme exhibited the highest EC affinity (K