Kuhn_1982_Biochem.Pharmacol_31_781

Guinea pig liver has the highest chloramphenicol-hydrolyzing capacity among the livers of various mammals. The enzyme responsible for the hydrolysis of the amide-bond in chloramphenicol is one of the isoenzymes of the microsomal nonspecific carboxylesterases. This isoenzyme is related to the well-known acetanilide-hydrolyzing carboxylesterases/amidases of pig and rat liver. The guinea pig liver enzyme is purified 24-fold starting with microsomes. The purified enzyme is essentially free from other proteins except other carboxylesterase isoenzymes with similar properties. The chloramphenicol-hydrolyzing esterase has an apparent molecular weight of about 180,000, a subunit weight of 60,000 and a pH optimum at 8.5. It also hydrolyzes methyl butyrate and acetanilide and it is completely inhibited by diethyl-4-nitrophenyl phosphate. Two assay procedures for the enzymatic chloramphenicol hydrolysis are described: a thin-layer chromatographic assay using radioactive chloramphenicol and a colorimetric assay utilizing the reaction of the liberated amine with trinitrobenzenesulfonic acid.