In thepresentstudy,proteaseproducing Bacillus subtilis GACAS8wasisolatedfrommarineascidian
Phallusia arabica and identified by16SrRNAgenesequence.Optimizationofcultureconditionfor
proteaseproductionusingagriculturalwasteshaddonebyresponsesurfacemethodology.Thestatistical
optimization studiesrevealedthattheproteaseproductionwashigherwhenthegroundnutoilcakeand
cabbage leafwereusedassubstrate.Theproteaseactivitywasoptimumat50 1C andpH9anditretained
more than60%ofactivity,evenatpH12andthemolecularweightofthepurified proteasewas41kDa.
The activityofthepurified proteasewasstimulatedbyMg2þ and Ca2þ and itwasresistanttoTween20,
Tween 40andSDS.TheproteaseactivitywasstronglyinhibitedinthepresenceofEDTA.Theremarkable
properties suchastemperatureandpHstabilityfoundwiththisenzymeassurethatitcouldbea
potentialcandidateforindustrialapplications.