AbstractBACKGROUNDBioremediation using enzymes has become an attractive approach for removing hazardous chemicals from the environment. The present study investigated the production, optimization and applications of an extracellular alkaline protease (AkP) from the edible mushroom Termitomyces clypeatus focusing on bioremediation.RESULTSFor optimized enzyme production, the variables significantly influencing the protease activity were screened through the 11 factorial Plackett–Burman design. The optimum values of the selected variables and their mutual interactions were determined through response surface methodology using the Box–Behnken experimental design. Overall a 10‐fold increase in enzyme activity (1830 U mL‐1) was obtained which closely matched the yield predicted by the statistical model (1778.63 U mL‐1) with R2 = 0.9819. AkP could efficiently dehair the goat skin, detached bird feather vanes from its shaft and reduce BOD, COD and pH of tannery waste effluent. The AkP also demonstrated bactericidal property against Staphylococcus aureus as detected by MIC and flow cytometry analyses.CONCLUSIONAs an ecofriendly alternative, the enzyme showed significant promise for bioremediation and industrial applications through time‐saving bioprocesses. This is first report of alkaline protease from T. clypeatus or from a fungal source with wide‐ranging application potential. © 2014 Society of Chemical Industry