Current Microbiology

, Volume 15, Issue 4, pp 203–205 | Cite as

Improved purification of α-amylase isolated fromRhizomucor pusillus by affinity chromatography

  • Sandra L. Turchi
  • Thomas Becker

Abstract

A highly purified extracellular α-amylase was isolated fromRhizomucor pusillus with minimum loss of enzymatic activity. The enzyme was purified from the mycelium-free liquid filtrate of the thermophilic moldRhizomucor pusillus. Maximum enzyme yields were attained after 5 days of growth on liquid starch-yeast extract at 45°C and pH 7.0. The crude enzyme preparation was first concentrated 80-fold by ultrafiltration. Purification was recently achieved with high-performance liquid chromatography and Waters Protein Pak 300 SW. Improved purification was then achieved with a dextrin-bound affinity column, with a 59-fold increase in specific activity from the crude enzyme preparation. This final enzyme preparation produced a single band on polyacrylamide gel electrophoresis. The molecular weight determined by SDS gel electrophoresis was 52,000 daltons.

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Copyright information

© Springer-Verlag New York Inc. 1987

Authors and Affiliations

  • Sandra L. Turchi
    • 1
  • Thomas Becker
    • 1
  1. 1.Department of ChemistryMillersville UniversityMillersvilleUSA

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