The N-benzoyl-L-tyrosine ethyl ester (BTEE) anchored in cyclodextrin (beta and beta-methyl CD) serves as an excellent substrate for subtilisin Carlsberg catalysis. The rate of hydrolysis was found to be approximately twofold higher than that of the methanolic substrate. The K-mapp and V-max values for the CD anchored substrates were significantly higher than the methanol-solubilized BTEE.