Enzyme purification by electrodecantation.
1. Electrodecantation has been applied to the initial fractionation from crude material of pig kidney beta-d-glucosidase and sheep testicular N-acetylglucosaminidase. 2. The isoelectric points determined by isoelectric focusing were pH4.9 for the beta-d-glucosidase and pH6.3 for the N-acetylglucosaminidase. 3. Electrodecantation of pig kidney extract and sheep testicular extract was carried out at pH4.9 and 6.3 respectively. 4. A six- to ten-fold increase in specific activity could be obtained with good recoveries after a single cycle of electrodecantation. 5. The technique has also been used to purify further an extracellular Bacillus subtilis protease preparation. 6. Attempts to use electrodecantation for the concentration of very dilute enzyme solutions resulted in considerable loss of activity. 7. The limitations and potential use of the technique in laboratory-scale enzyme preparation are discussed.
D000179:Acrylates; D000818:Animals; D001412:Bacillus subtilis / Q000201:enzymology; D003956:Dialysis; D004586:Electrophoresis; D004798:Enzymes / Q000302:isolation & purification*; D005782:Gels; D005959:Glucosidases / Q000302:isolation & purification; D006596:Hexosaminidases / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology; D008297:Male; D008722:Methods; D010447:Peptide Hydrolases / Q000302:isolation & purification; D012756:Sheep; D013552:Swine; D013737:Testis / Q000201:enzymology
D000179:Acrylates; D004798:Enzymes; D005782:Gels; D005959:Glucosidases; D006596:Hexosaminidases; D010447:Peptide Hydrolases