ISSN 1995-5537

«Біотехнологія» journal V. 5, No. 5, 2012
Р. 91-99, Bibliography 26, Russian.
Universal Decimal classification: 577.152.32

PURIFICATION OF Aspergillus flavus var. oryzae AND Bacillus subtilis α-AMYLASES ABD THEIR PROPERTIES

K. V. Avdiyuk, L. D. Varbanets, L. A. Safronova, E. S. Harkevich

Institute of Microbiology and Virology of National Academy of Sciences of Ukraine, Kyiv

Methods of isolation and purification of enzymes with α-amylase activity of the two producers — Aspergillus flavus var. oryzae 80428 and Bacillus subtilis 147 were selected which included: ammonium sulfate fractionation, gel filtration on TSK-gel Toyopearl HW-50 and ionexchange chromatography on TSK-gel DEAEToyopearl 650 M (for α-amylase, A. flavus var. oryzae); fractionation by ammonium sulfate and affinity sorption on starch (for α-amylase, B. subtilis 147). α-Amylase of A. flavus var. оryzae was purified in 37 times and α-amylase B. subtilis 147 — in 20 times in comparison with enzyme activity in supernatant of cultural medium. α-Amylase of A. flavus var. oryzae showed maximum activity at 60 °С and pH 6,0 and retained 100% activity after 24 h at pH 5,0–7,0 and after 3 h at 37 °С. The pH and temperature for optimum activity of the enzyme were 90 °С and 8,0. It was stable at рН 7,0–9,0 after 24 h and at 37 °С, 60 °С, 70 °С after 3 h and retained 80% of the initial activity after 3 h at 80 °С, respectively.

Key words: ?-amylase, gel-filtration, іоnexchange chromatography, Aspergillus flavus var. oryzae, Bacillus subtilis, purification.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008