一株酸性糖化酶的分离纯化及酶学性质研究.pdfVIP

现代食品科技 Modern Food Science and Technology 2012, Vol.28, No.10 一株酸性糖化酶的分离纯化及酶学性质研究 朱婧，王青艳，廖思明，秦艳，申乃坤，黄日波 (广西科学院国家非粮生物质能源工程技术研究中心，广西南宁 530007) 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(NH ) SO 沉淀、Sepharose HP阴离子交换层析、 4 2 4 Sephacryl S-200层析柱分离纯化，SDS电泳测定其分子量为120 kDa。该酶最适作用温度为65 ℃；酶的最适反应pH值为4.0；在 2+ 2+ pH 2.2~7.6之间，具有较好的酸稳定性；酶的Km值为0.94 mg/mL，Vmax=142.43 mol(Glu)/min·L。Cu 和Co 对酶活有较强的促进作 用，10 mM 的Cu2+使酶活力提高到129%，Fe3+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pH 4.0 ，50 ℃反应 1h，生淀粉酶活力为0.39 U，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，酶学性质 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。 关键词：酸性；糖化酶；黑曲霉；分离纯化 文章篇号：1673-9078(2012)10-1294-1297 Purification of an Acidic Glucoamylase from Aspergillus niger and its Enzymatic Properties ZHU Jing, WANG Qing-yan, LIAO Si-ming, QIN Yan, SHEN Nai-kun, HUANG Ri-bo (National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences , Nanning 530007, China) Abstract: 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 SDSwas 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 of pH 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