Purification of alpha-amylases using magnetic alginate beads.
Magnetic alginate beads were used to purify alpha-amylases from porcine pancreas, starchzyme, BAN 240L (a commercial purification from Bacillus subtilis), and wheat germ. The beads bound a significant level of alpha-amylase activity from porcine pancreas, BAN 240L, and wheat germ. In each case, the enzyme activity could be eluted by using 1.0 M maltose, a known competitive inhibitor of alpha-amylase. In the case of BAN 240L, 3.6-fold purification with 72% recovery of activity was observed. In the case of wheat germ enzyme, starting from the crude extract, 48-fold purification with 70% activity recovery was observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis also indicated considerable purification in the latter case.
Teotia
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Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas.
Gupta
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D000464:Alginates / Q000737:chemistry / Q000648:ultrastructure; D000681:Amylases / Q000737:chemistry / Q000302:isolation & purification; D001671:Biochemistry / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D052203:Ferrosoferric Oxide; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D007501:Iron / Q000737:chemistry; D008024:Ligands; D008280:Magnetics; D008855:Microscopy, Electron, Scanning; D010087:Oxides / Q000737:chemistry; D011485:Protein Binding; D013997:Time Factors; D014908:Triticum / Q000201:enzymology; D000516:alpha-Amylases / Q000737:chemistry* / Q000302:isolation & purification*
D000464:Alginates; D006603:Hexuronic Acids; D008024:Ligands; D010087:Oxides; D020723:Glucuronic Acid; D007501:Iron; D000681:Amylases; D000516:alpha-Amylases; D052203:Ferrosoferric Oxide