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Purification and characterization of α-amylase fromAspergillus flavus

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Abstract

Aspergillus flavus produced approximately 50 U/mL of amylolytic activity when grown in liquid medium with raw low-grade tapioca starch as substrate. Electrophoretic analysis of the culture filtrate showed the presence of only one amylolytic enzyme, identified as an α-amylase as evidenced by (i) rapid loss of color in iodine-stained starch and (ii) production of a mixture of glucose, maltose, maltotriose and maltotetraose as starch digestion products. The enzyme was purified by ammonium sulfate precipitation and ion-exchange chromatography and was found to be homogeneous on sodium dodecyl sulfate— polyacrylamide gel electrophoresis. The purified enzyme had a molar mass of 52.5±2.5 kDa with an isoelectric point at pH 3.5. The enzyme was found to have maximum activity at pH 6.0 and was stable in a pH range from 5.0 to 8.5. The optimum temperature for the enzyme was 55°C and it was stable for 1 h up to 50°C. TheKm andV for gelatinized tapioca starch were 0.5 g/L and 108.67 μmol reducing sugars per mg protein per min, respectively.

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Authors and Affiliations

  1. School of Biological Sciences, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    S. L. Khoo, A.-A. Amirul, M. Kamaruzaman, N. Nazalan & M. N. Azizan

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  1. S. L. Khoo
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  2. A.-A. Amirul
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  3. M. Kamaruzaman
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  4. N. Nazalan
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  5. M. N. Azizan
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Khoo, S.L., Amirul, AA., Kamaruzaman, M. et al. Purification and characterization of α-amylase fromAspergillus flavus . Folia Microbiol 39, 392–398 (1994). https://doi.org/10.1007/BF02814445

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  • DOI: https://doi.org/10.1007/BF02814445

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