Purification, partial characterization, and covalent immobilization-stabilization of an extracellular α-amylase from Aspergillus niveus
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- alfa-amylasy izolace a purifikace metabolismus MeSH
- amylopektin metabolismus MeSH
- amylosa metabolismus MeSH
- Aspergillus enzymologie MeSH
- chromatografie iontoměničová MeSH
- chromatografie na tenké vrstvě MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- enzymy imobilizované chemie metabolismus MeSH
- gelová chromatografie MeSH
- glykogen metabolismus MeSH
- hydrolýza MeSH
- koncentrace vodíkových iontů MeSH
- molekulární sekvence - údaje MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alfa-amylasy MeSH
- amylopektin MeSH
- amylosa MeSH
- enzymy imobilizované MeSH
- glykogen 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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