Preparation of biocatalytic nanofibres with high activity and stability via enzyme aggregate coating on polymer nanofibres
Preparation of biocatalytic nanofibres with high activity and stability via enzyme aggregate coating on polymer nanofibres
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DOI：
10.1088/0957-4484/16/7/011
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发表时间：
2005-07-01
期刊：
影响因子：
3.5
通讯作者：
Gu, MB
中科院分区：
文献类型：
--
作者：
Kim, BC;Nair, S;Gu, MB
我们已经开发出一种独特的方法，用于在静电纺丝聚合物纳米纤维表面上制造酶聚集体涂层。该方法采用将种子酶共价连接到由聚苯乙烯和聚（苯乙烯-共-马来酸酐）的混合物组成的纳米纤维上，然后进行戊二醛（GA）处理，所述戊二醛处理将另外的酶分子交联并从溶液聚集到共价连接的种子酶分子上。这些交联的酶聚集体通过种子酶分子的接头共价连接到纳米纤维，预期由于增加的酶负载而提高酶活性，并且还提高酶稳定性。为了证明这一原理，我们将α-糜蛋白酶（CT）涂覆在由聚苯乙烯和聚（苯乙烯-马来酸酐共聚物）的混合物电纺的纳米纤维上。CT聚集体涂覆的纳米纤维的初始活性比仅具有一层共价连接的CT分子的纳米纤维高9倍。CT聚集体涂覆的纳米纤维的酶稳定性大大提高，在严格的振荡条件下观察一个月基本上没有可测量的活性损失。这种将酶涂覆在纳米纤维上的新方法，产生高活性和稳定性，创造了一种有用的新的生物催化固定化酶系统，在生物转化、生物修复和生物传感器中具有潜在的应用。
We have developed a unique approach for the fabrication of enzyme aggregate coatings on the surfaces of electrospun polymer nanofibres. This approach employs covalent attachment of seed enzymes onto nanofibres consisting of a mixture of polystyrene and poly(styrene-co-maleic anhydride), followed by a glutaraldehyde (GA) treatment that cross-links additional enzyme molecules and aggregates from the solution onto the covalently attached seed enzyme molecules. These cross-linked enzyme aggregates, covalently attached to the nanofibres via the linkers of seed enzyme molecules, are expected to improve the enzyme activity due to increased enzyme loading, and also the enzyme stability. To demonstrate the principle, we coated a-chymotrypsin (CT) on nanofibres electrospun from a mixture of polystyrene and poly(styrene-co-maleic anhydride). The initial activity of CT-aggregate-coated nanofibres was nine times higher than nanofibres with just a layer of covalently attached CT molecules. The enzyme stability of CT-aggregate-coated nanofibres was greatly improved with essentially no measurable loss of activity over a month of observation under rigorous shaking conditions. This new approach of enzyme coating on nanofibres, yielding high activity and stability, creates a useful new biocatalytic immobilized enzyme system with potential applications in bioconversion, bioremediation, and biosensors.