Purification and characterization of amylase produced by Aspergillus oryzae FAFU from rice wine fermentation starter
The crude amylase was extracted from Aspergillus oryzae FAFU.The enzymes were purified by precipitation through ammonium sulfate,desalted by dialysis,and successively purified by chromatography on DEAE Sepharose and Sephacryl S-200 column to produce three amylase fractions termed Amy-Ⅰ,Amy-Ⅱ,and Amy-Ⅲ,respectively.Amy-Ⅰ and Amy-Ⅱ were identified as o-amylase,Amy-Ⅲ was identified as glucoamylase according to the results of TLC analysis for their enzymatic hydrolysates,and their molecular mass were about 48.6,49.7,93.5 kDa respectively through SDS-PAGE.Some enzymatic properties of Amy-Ⅰ and Amy-Ⅱ were characterized in this research.The optimal reaction temperature of Ant-Ⅰ was determined to be 50 ℃.It was stable when the temperature was lower than 50 ℃,and its half-life was 40 min at 55 ℃.The optimal reaction pH was 6.0,and its pH stabilities were kept in the range 7.2-9.6.The optimal reaction temperature of Amy-Ⅱ was determined to be 50 ℃,it was stable when the temperature lower than 45 ℃ and its half-life was 30 min at 50℃.Its optimal reaction pH was 6.6,and it kept stable in the pH range 8.6-10.0.
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