Purification of ram seminal plasma acid alpha-glucosidase.
1. Ram seminal plasma alpha-glucosidase has been purified in order to increase our knowledge of this enzyme and of its role in epididymal physiology. 2. Since the enzyme behaved differently from other known acid alpha-glucosidases and was not affinity-adsorbed on dextran gels, another approach had to be used. 3. The final procedure included an ethanol precipitation step, sequential chromatography on hydroxylapatite and DEAE Sepharose CL-6B, isoelectric focusing in polyacrylamide gels and ultrafiltration. 4. The resulting purification factor of alpha-glucosidase was 9822 with an overall yield of 5%. 5. The purified material consisted of several isoforms with a mol. wt of 105,000 and isoelectric points varying between 4 and 5.
D000818:Animals; D011232:Chemical Precipitation; D002845:Chromatography; D005087:Glucan 1,4-alpha-Glucosidase / Q000302:isolation & purification*; D005959:Glucosidases / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008297:Male; D008970:Molecular Weight; D012661:Semen / Q000201:enzymology*; D012756:Sheep; D014462:Ultrafiltration
D005959:Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase