Purification and characterization of rat epididymal neutral beta-galactosidase and its changes during in vivo development.
A neutral beta-D-galactosidase has been partially purified from rat epididymis and characterized. The enzyme having molecular mass of approximately 50 kilodaltons has been purified 400-fold by using calcium phosphate gel adsorption, DEAE-cellulose chromatography, Sephadex G-100 gel filtration, and concanavalin A--agarose affinity chromatography. Although the neutral enzyme binds to the concanavalin A affinity column, the activity could be eluted with alpha-methyl mannoside only if the buffer contained salt (NaCl) at a concentration as high as 0.3 M. The enzyme was of cytosolic origin, since 90% of the total enzymic activity of the tissue homogenate was recovered in the soluble fraction of these cells. The neutral beta-galactosidase was not dependent on metal ions for its activity and it had a pH optimum of 7.0. Zn2+, p-chloromercuribenzoate, Hg2+, and Pb2+ served as potent inhibitors of the enzyme. There was a marked increase (approximately fourfold) in the specific activity of the neutral beta-galactosidase during sexual maturity of epididymis in vivo.
Dutta
P
P
Indian Institute of Chemical Biology, Jadavpur, Calcutta.
Majumder
G C
GC
D000818:Animals; D002451:Cell Compartmentation; D002458:Cell Fractionation; D003600:Cytosol / Q000201:enzymology; D004822:Epididymis / Q000201:enzymology* / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D008297:Male; D051381:Rats; D013379:Substrate Specificity; D013997:Time Factors; D015032:Zinc / Q000494:pharmacology; D001616:beta-Galactosidase / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
D001616:beta-Galactosidase; D015032:Zinc