Purification, partial characterization, and covalent immobilization-stabilization of an extracellular α-amylase from Aspergillus niveus
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- alpha-Amylases isolation & purification metabolism MeSH
- Amylopectin metabolism MeSH
- Amylose metabolism MeSH
- Aspergillus enzymology MeSH
- Chromatography, Ion Exchange MeSH
- Chromatography, Thin Layer MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Enzymes, Immobilized chemistry metabolism MeSH
- Chromatography, Gel MeSH
- Glycogen metabolism MeSH
- Hydrolysis MeSH
- Hydrogen-Ion Concentration MeSH
- Molecular Sequence Data MeSH
- Amino Acid Sequence MeSH
- Sequence Alignment MeSH
- Enzyme Stability MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- alpha-Amylases MeSH
- Amylopectin MeSH
- Amylose MeSH
- Enzymes, Immobilized MeSH
- Glycogen MeSH
An extracellular amylase secreted by Aspergillus niveus was purified using DEAE fractogel ion exchange chromatography and Sephacryl S-200 gel filtration. The purified protein migrated as a single band in 5 % polyacrylamide gel electrophoresis (PAGE) and 10 % sodium dodecyl sulfate (SDS-PAGE). The enzyme exhibited 4.5 % carbohydrate content, 6.6 isoelectric point, and 60 and 52 kDa molar mass estimated by SDS-PAGE and Bio-Sil-Sec-400 gel filtration column, respectively. The amylase efficiently hydrolyzed glycogen, amylose, and amylopectin. The end-products formed after 24 h of starch hydrolysis, analyzed by thin layer chromatography, were maltose, maltotriose, maltotetraose, and maltopentaose, which classified the studied amylase as an α-amylase. Thermal stability of the α-amylase was improved by covalent immobilization on glyoxyl agarose (half-life of 169 min, at 70 °C). On the other hand, the free α-amylase showed a half-life of 20 min at the same temperature. The optima of pH and temperature were 6.0 and 65 °C for both free and immobilized forms.
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