Purification of α-amylases using magnetic alginate beads
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Magnetic alginate beads were used to purify α-amylases from porcine pancreas, starchzyme, BAN 240L (a commercial purification from Bacillus subtilis), and wheat germ. The beads bound a significant level of α-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 α-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.
Index EntriesAffinity separation α-amylase magnetic alginate beads macroaffinity ligands wheat amylase
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- 2.Safarik, I. and Safarikova, M. (1997), in Scientific and Clinical Applications of Magnetic Carriers, Hafeli, U., Schut, W., Teller, J., and Dorowski, M., eds., Plenum, New York, pp. 323–340.Google Scholar
- 4.Worlock, A. J., Sidgwick, A., Horsburgh, T., and Bell, P. R. L. (1991), Biotechniques 10, 310–315.Google Scholar
- 7.Walsh, G. and Headon, D. (1994) in Protein Biotechnology, Wiley, Chichester, pp. 321–322.Google Scholar
- 9.Decker, L. A. (1977), Worthington Enzyme Manual, Worthington Biochemical, Freehold, NJ.Google Scholar
- 11.Hames, B. D. (1986), in Gel Electrophoresis of Protein: A Practical Approach, Hames B. D. and Rickwood, D., eds., IRL, Oxford, pp. 1–86.Google Scholar
- 12.Rozie, H., Somers, W., Bonte, A., Visser, J., van’t Riet, J., and Rombouts, F. (1988), Biotechnol. Appl. Biochem. 10, 346–358.Google Scholar
- 14.Rozie, H., Somers, W., Bonte, A., Visser, J., Van’t Riet, K., and Rombouts, F. (1991), Biotechnol. Appl. Biochem. 13, 181–195.Google Scholar