Enzyme immobilization has been used in non-aqueous enzymatic catalysis due to its advantages like enhancing stability,activity,and recycle rate.Thus,in the work,we mainly used molecular dynamics(MD)simulation and protein structure network(PSN)to study subtilisin Carlsberg(SC)immobilized onto carbon nanotube(CNT)in water,acetonitrile and heptane solvents,in order to explore activation mechanism of enzymes in non-aqueous media.