In order to separate lumbrokinase in Pheretima praepinguis and to study the enzymatic properties, with Pheretima praepinguis as material, lumbrokinase was separated with the salting out method. Lumbrokinase activity was measured with casein medium plate method. The effect of pH and temperature on lumbrokinase activity was studied. The activity of lumbrokinase separated from Pheretima praepinguis was relatively high. Lumbrokinase activity in neutral or slightly alkaline environment was higher. Lumbrokinase had tolerance ability to high temperature, with highly enzymatic activity under 60° and wide range of temperature adaptation.