Abstract
A rapid and efficient method the exploiting affinity of α-amylase for its substrate starch is described. α-amylase from Bacillus licheniformis was purified to homogeneity by ammonium sulphate precipitation and affinity chromatography with 230-fold purification. The α-amylase adsorption to various starches was examined in order to screen its ability for highest binding to starch. The α-amylase was bound to starch more tenaciously, hence various eluants like maltose, soluble starch and high salts could not elute the bound α-amylase. However, the bound α-amylase was instantly eluted using 2% (w/v) dextrin. The purified enzyme showed a single polypeptide on SDS-PAGE, with a molecular weight of 58 kD. Western blot analysis confirmed the specificity of antibody raised against purified α-amylase.
Original language | English (US) |
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Pages (from-to) | 371-375 |
Number of pages | 5 |
Journal | World Journal of Microbiology and Biotechnology |
Volume | 21 |
Issue number | 3 |
DOIs | |
State | Published - Apr 2005 |
Externally published | Yes |
Keywords
- Affinity adsorption
- Bacillus licheniformis
- α-amylase
ASJC Scopus subject areas
- Biotechnology
- Physiology
- Applied Microbiology and Biotechnology