Affinity Chromatography of Rabbit Liver β-Glucuronidase

Abstract

An affinity adsorbent was prepared by coupling p-aminophenyl 1-thio-β-D-gluco-pyranosiduronic acid with CH-Sepharose 4B. β-Glucuronidase from rabbit liver was adsorbed on p-aminophenyl 1-thio-β-D-glucopyranosiduronic acid-CH-Sepharose then eluted with basic buffer or salt solution. The specific enzyme activity increased 46-fold and essentially all of the activity of the enzyme was recovered in elution buffer containing 0.1M NaCl. N-Acetyl-β-glucosaminidase was adsorbed slightly but β-glucosidase and β-galactosidase were not adsorbed on the affinity adsorbent. The adsorbent retained affinity for the enzyme after being used ten times.