Purification of wheat germ amylase by precipitation.
alpha-Amylase from various sources was found to bind alginate in free solution. The alginate-enzyme complex could be precipitated with Ca(2+). The enzyme activity could be recovered by dissolving the precipitate in 1 M maltose and precipitating alginate alone by addition of Ca(2+). Based upon these observations, alpha-amylase from wheat germ was purified with 68-fold purification and 72% recovery. The molecular weight estimated by SDS-PAGE was 18 kDa. The method also worked equally well with alpha-amylase for the whole wheat seed. The latter enzyme could be purified 54-fold with 70% activity recovery. The molecular weight of this second enzyme was estimated to be 45 kDa by SDS-PAGE.
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Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi, 110016, India.
Sharma
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Gupta
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D000464:Alginates; D002118:Calcium; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D008320:Maltose; D008970:Molecular Weight; D012639:Seeds / Q000201:enzymology; D014908:Triticum / Q000201:enzymology*; D000516:alpha-Amylases / Q000737:chemistry / Q000302:isolation & purification*
D000464:Alginates; D006603:Hexuronic Acids; D008320:Maltose; D020723:Glucuronic Acid; D000516:alpha-Amylases; D002118:Calcium