交联牛肝β-糖苷酶聚合物的制备、表征和应用.PDF

现代食品科技 Modern Food Science and Technology 2015, Vol.31, No.10 交联牛肝β-糖苷酶聚合物的制备、表征及应用 秦烨芝，叶敏，李宁，宗敏华 （华南理工大学轻工与食品学院制浆造纸工程国家重点实验室，广东广州 510640） 摘要：牛肝β-糖苷酶是一种能催化核苷类化合物区域选择性糖基化反应合成二糖核苷的生物催化剂。在已有的研究报道中，采 用的生物催化剂都是游离酶，不利于其大规模应用。交联酶聚合物(CLEAs)作为一种新型的无载体固定化酶，具有制备简便、成本低、 体积酶活高等优点。本文研究牛肝β-糖苷酶CLEAs 的制备，并对其酶学性质和结构进行了研究，探讨了其在二糖核苷合成中的应用。 最适的沉降剂、交联剂及交联时间分别为70%饱和度的(NH ) SO 、50% (V/ V)葡聚糖多醛及8 h。在最适条件下，CLEAs 的活性回收 4 2 4 率为58%。该CLEAs 的最适pH 、温度、V 及K 分别为8.5、60 ℃、0.13 μmol/(min mg)及1.43 mM。并且该CLEAs 具有较好的操 max m 作稳定性，重复利用6 批次后，其催化合成5’-O-β-葡萄糖基-2’-脱氧尿苷的相对产率仅下降20%。牛肝β-糖苷酶CLEAs 呈无定形态。 关键词：β-糖苷酶；交联酶聚合物；二糖核苷 文章篇号：1673-9078(2015)10-210-214 DOI: 10.13982/j.mfst.1673-9078.2015.10.035 Preparation, Characterization, and Application of Cross-linked Enzyme Aggregates ofβ-glycosidases from Bovine Liver QIN Ye-zhi, YE Min, LI Ning, ZONG Min-hua (State Key Laboratory of Pulp and Paper Engineering, College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China) Abstract: β-glycosidase from bovine liver is a biocatalyst that can catalyze the synthesis of disaccharide nucleosides via regioselective glycosylation of nucleosides. In previous reports, the used biocatalysts were free enzymes, which limit the large-scale applications. As novel carrier-free immobilized enzymes, cross-linked enzyme aggregates (CLEAs) have many advantages, such as easy preparation, low cost, and high enzyme volumetric activities. CLEAs of β-glycosidase from bovine liver were prepared, their enzymatic properties and structures were studied, and their application for the syn