title: Improved properties of bovine erythrocyte acetylcholinesterase, isolated by papain cleavage.
abstract: A simple and rapid procedure involving papain cleavage of the membrane anchor was used to isolate membrane-bound acetylcholinesterase from bovine erythrocytes. The solubilized enzyme was purified 930-fold by ion exchange chromatography and gel filtration. The properties of the papain-cleaved acetylcholinesterase were compared with those of a commercial acetylcholinesterase, solubilized from the erythrocyte membranes by detergents. Cleavage of the membrane anchor eliminated dimer aggregation, caused a pH shift in thermal stability and resulted in increased stability in organic solvents. Bovine serum albumin, used as stabilizer of the commercial enzyme preparation, increased the thermal stability but concomitantly decreased the activity of acetylcholinesterase at pH 6-8. The improved stability of the cleaved acetylcholinesterase, especially in organic solvents, may enhance the biosensor performance of the enzyme.
authors.affiliation: Department of Enzyme Technology, GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Schmidt-Dannert
authors.forename: H M
authors.initials: HM
authors.lastname: Kalisz
authors.forename: I
authors.initials: I
authors.lastname: Safarík
authors.forename: R D
authors.initials: RD
authors.lastname: Schmid
mesh_terms: D000110:Acetylcholinesterase / Q000097:blood* / Q000737:chemistry; D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010206:Papain / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology
chemical_list: D002800:Cholinesterase Inhibitors; D012710:Serum Albumin, Bovine; D000110:Acetylcholinesterase; D010206:Papain