β-淀粉酶的活性和热稳定性研究.pdfVIP

β-淀粉酶的活性和热稳定性研究 田辉，张红，张剑∗ （武汉工业学院化学与环境工程系，湖北武汉 430023 ） 摘要 本文借助 DNS 试剂并通过分光光度法建立了在底物存在条件下 β-淀粉酶的活性 测定方法。并用该法从温度、淀粉浓度以及钙的添加量等方面探讨了 β-淀粉酶的活性和 热稳定性。研究表明，该酶作用底物的最适酸度为 4.5 ，钙离子的添加量直接关系到 β- 淀粉酶的活性和热稳定性。当向 2% （v/v ）初始浓度的淀粉溶液中加入 34 μmol CaCl2 / μg 酶蛋白量时，可使该酶的热稳定性和催化效率达到最大，在 30℃和pH 4.5 的条件下， 该酶的催化活性是对照酶的 1.25 倍。由此可见，向酶水解淀粉的反应液中添加适量的 2+ Ca ，可提高 β-淀粉酶的利用率。 关键词 β-淀粉酶；酶活力；Ca2+离子；热稳定性酶 Studies on Thermal Stability and Activity of β-Amylase TIAN, Hui ZHANG, Hong ZHANG, Jian* ( Department of Chemistry and Environmental Engineering, Wuhan Polytechnic University, ChangQing Garden, Hankou, Wuhan, 430023 ) Abstract A determination method for activity of β-amylase in presence of starch had been established by spectrophotometry in accordance with DNS procedure. Thermal stability and activity of β-amylase were studied from the aspects of temperatures, starch concentrations 2+ and additions of Ca in this paper. The results indicated that β-amylase displayed a pH 2+ activity optimum of 4.5 and addition of Ca was directly related to catalytic efficiency and thermostability of β-amylase. When addition of CaCl2 was 34 μmol / μg enzyme protein, thermostability and activity of the enzyme in the presence of starch achieved maximum values. Catalytic efficiency of β-amylase incubated at 34 μmol CaCl2 / μg enzyme protein solution (30oC, pH 4.5) was approximately 1.25-fold as the control enzyme during starch hydrolysis reaction. Therefore