机构地区: 广西科学院
出 处: 《现代食品科技》 2012年第10期1294-1297,1406,共5页
摘 要: 从土壤中筛选得到一株产酸性糖化酶的黑曲霉ASP-S21,粗酶液通过(NH4)2s04沉淀、Sepharose HP阴离子交换层析、SephacrylS-200层析柱分离纯化,SDS-PAGE电泳测定其分子量为120kDa。该酶最适作用温度为65℃;酶的最适反应pH值为4.0;在pH2.2-7.6之间,具有较好的酸稳定性;酶的Km值为0.94mg/mL,Vmax=142.43mol(Glu)/min-L。Cu2+和Co2+对酶活有较强的促进作用,10mM的Cu2镟酶活力提高到129%,Fe^3+对酶催化活力抑制作用较强。该酶且具有部分降解生淀粉的能力,在pH4.0,50℃反应1h,生淀粉酶活力为0.39U,RDA值为4.57%。得到的黑曲霉ASP-S21酸性糖化酶,产生的糖化酶活力高、耐酸稳定性好,酶争陛质符合淀粉糖化工业化过程中对酶的要求,具有良好的研究前景。 An acidic glucoamylase producing strain Aspergillus niger ASP-S21 was isolated from soil. Glucoamylase was purified to homogeneity by ammonium sulfate precipitation ,ion-exchange chromatography with a Sepharose HP column ,and column chromatography with a Sephacryl S-200 column, the molecular weight of SDS-PAGE was 120 kDa. The study of its enzymatic properties showed that the optimum temperature and pH for glucoamylase of such strain were 65℃ and 4.0, respectively. Besides, it had good acid stability within the range ofpH value from 2.2 to7.6. The Km for glucoamylase was calculated to be 0.94 mg/mL, and the Vmax was 142.43 mol(Glu)/min.L. The enzyme activity was enhanced by Cu2+and Co2+, and the 10 mM of Cu2+ could improve the glucoamylase activity to 129%. However, the enzyme was strongly inhibited by Fe3+. Its RSGA activities were 0.39U and the RDA values were 4.57%. The glucoamylase was suitable for applying in starch sugar industry.