交联β-葡萄糖苷酶聚集体的制备及其性质-食品科学.pdf

254 2007, Vol. 28, No. 12 食品科学 ※生物工程 罗建平，欧 杰，潘利华 (合肥工业大学生物与食品工程学院，安徽 合肥 230009) ：交联酶聚集体法是一种新型的无载体酶固定化方法。本实验用该法固定化黑曲霉β- 葡萄糖苷酶，对其 制备条件、结构特征、酶学性质进行了研究。结果表明，β-葡萄糖苷酶酶液在冰水浴中经90%的硫酸铵沉淀30min 后，以5% 戊二醛水溶液进行酶聚集体交联反应，可获得较高活性的交联酶聚集体，酶活回收率达75%，对对- 硝基苯基-β-D-葡萄糖苷作用的最适温度、最适pH值和K － 3 m 值分别为60℃、pH4.0和4.61×10 mol/L，与游离酶 相比，表现更高的底物亲和力。将其用于大豆异黄酮活性苷元染料木素的合成，40min 内对染料木苷转化率达到 6 6 % 。 ：β- 葡萄糖苷酶；交联酶聚集体；无载体固定化；大豆异黄酮；应用 Preparation and Properties of Cross-linked β-glucosidase Aggregates LUO Jian-ping，OU Jie，PAN Li-hua (School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009, China) Abstract：Cross-linked enzyme aggregates (CLEAs) were used to immobilize β-glucosidase from Aspergillus niger. β- glucosidase was precipitated by 90% ammonium sulfate solution for 30 min at 0 ℃, then the aggregates were cross-linked by 5% glutaraldehyde solution. The obtained cross-linked β-glucosidase aggregates remained 75% of total enzyme activity. Its optimum －3 reaction temperature, reaction pH and Km to pNPG were 60 ℃, 4.0 and 4.61×10 mol/L, respectively. Compared to the free β-glucosidase, the cross-linked β-glucosidase aggregates had higher affinity to the substrate. The conversion rate was 66% when cross-linked β-glucosidase aggregates were employed to genistein production from soybean isoflavone genistin in 40 min. Key words：β-glucosidase；cross-linked enzyme aggregates；carrier-free immobilization；soybean isoflavones； application 中图分类号：Q814.2 文献标识码：A 文章编号：1002-6630(2007)12-02