A single-step purification of rat pancreatic and salivary amylase by affinity chromatography

✍ Scribed by Peter H. Burrill; Patsy M. Brannon; Norman Kretchmer

• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 263 KB
• Volume: 117
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90798-3

✦ Synopsis

Rat pancreatic and salivary amylase [( 1 -4) a-D-gkan glucanohydrolase] (EC 3.2.1.1) were purified 20-to 50-fold with a yield above 75% by a single-step affinity chromatographic procedure. The affinity ligand was an a-glucohydrolase inhibitor (proprietary name Bay g5421) coupled to w-aminohexyl-Sepharose 4B. Pancreatic amylase was eluted as a single peak at pH 7.4 with 0.1% glycogen or at pH 5.8 without glycogen. Salivary amylase could be eluted only with 0.1% glycogen containing buffer at pH 7.4. Pancreatic amylase migrated as a single homogeneous band on sodium dodecyl sulfate polyacrylamide gels, whereas salivary amylase migrated as a doublet.