Precepitation and purification of amylase enzyme produced by streptomyces aureofaciens 77

  • Ibrahim, A.N. (Botany Department, Faculty of Agriculture, Al-Azhar University) ;
  • Ahmed, F.H. (Mycology Department, National Organization for Drug Control and Research) ;
  • Ibrahim, M.M.K. (Mycology Department, National Organization for Drug Control and Research) ;
  • Arafa, M.A.I. (Mycology Department, National Organization for Drug Control and Research)
  • Published : 1990.03.01

Abstract

Precipitation and purification of amylase secreted by Streptomyces aureofaciens 77 in liquid inorganic salts-starch medium under the optimum conditions were carried out. Ammonium sulphate fractionation was used to precipitate amylase in cell free culture filtrate. (NH/sub 4/)/sub 2/ SO/sub 4/ at a concentration of 50-70% saturation gave the highest enzyme yield. The obtained precipitates were redissolved in phosphate buffer (pH 7.0) and subjected to dialysis. The dialyzed enzyme preparation was applied to DEAE-cellulose column chromatography which resulted in an increase of purification up to 59.48 fold. A further step of purification was done by applying the obtained purified sample to Sephadex-G200 column chromatography which resulted in ann increase of purification up to 73. 92 fold. The results clearly indicated that the isolated amylase from S. aureofaciens 77 was only on type.

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