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

Publisher Website
Google Scholar
Abstract
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 was developed. It yields a stable folate-free preparation in quantity. The prufication steps included extraction of conjugase from crude lyophilized chicken pancreas in mercaptoethanol followed by DEAE-cellulose chromatography. The enzyme was further purified by absorption to alumina-C-.gamma.-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 2 peaks on Sephadex G-75 chromatography corresponding to MW of 50,000 and 25,000, respectively. The optimum temperature for the conjugase was between 35 and 50.degree., and the optimum pH was 8.5.

This publication has 2 references indexed in Scilit: