Partial Purification and Some Properties of Pteroylpolyglutamate Hydrolase (Conjugase) from Chicken Pancreas

A simple and rapid procedure for the purification of pteroylpolyglutamate hydrolase (conjugase) from chicken pancreas for the purpose of standardization of microbiological assays of folates has been developed. It yields a stable folate-free preparation in quantity. The purification steps included extraction of conjugase from crude lyophilized chicken pancreas in mer-captoethanol followed by DEAE-cellulose chromatography. The enzyme was further purified by absorption to alumina-C-γ-gel from which it was eluted with phosphate buffer, pH 6.5. The enzyme was concentrated by dialysis against 20 vol of 2 M sucrose, lyophilized, and vacuum-sealed. The purified conjugase exhibited two peaks on Sephadex G-75 chromatography corresponding to molecular weights of 50,000 and 25,000, respectively. The optimum