The
enzyme hydrolyses the substrate acetylthiocholine resulting in the
product thiocholine which reacts with Ellman’s reagent (DTNB) to
produce 2-nitrobenzoate-5-mercaptothiocholine and 5-thio-2-nitrobenzoate
which can be detected at 412 nm. 50 mM Tris–HCl
pH 8.0 was used as a buffer throughout the experiment unless
otherwise stated.