Preparation of a very stable immobilized biocatalyst of glucose oxidase from Aspergillus niger
Preparation of a very stable immobilized biocatalyst of glucose oxidase from Aspergillus niger
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DOI：
10.1016/j.jbiotec.2005.07.014
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发表时间：
2006-01-24
影响因子：
4.1
通讯作者：
Fernández-Lafuente, R
中科院分区：
文献类型：
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作者：
Betancor, L;López-Gallego, F;Fernández-Lafuente, R
葡萄糖氧化酶（GOX）被固定在不同的活化载体上，包括乙醛酰琼脂糖、环氧树脂珠和戊二醛活化的载体。用戊二醛预活化的载体上的固定化使得GOX的制备最耐热。因此，由于戊二醛化学给酶的高稳定性，我们提出了另一种技术，通过戊二醛提高多点连接：酶离子吸附在阳离子支持伯氨基，然后固定化制剂用戊二醛溶液处理。酶活性下降<20%。根据这种方法，我们实现了所有分析的固定化系统的最高稳定性，显示出比可溶性酶高100倍的半衰期。此外，该衍生物在有机溶剂（如甲醇）或环氧氢存在下显示出比离子吸附酶或可溶性酶更高的稳定性。因此，GOX胺化阳离子载体上的吸附和随后的处理与戊二醛作为一个非常成功的方法，实现了一个非常稳定的生物催化剂。(c)2005 Elsevier B.V.保留所有权利。
Glucose oxidase (GOX) has been immobilized on different activated supports, including glyoxyl agarose, epoxy sepabeads and glutaraldehyde-activated supports. Immobilization onto supports pre-activated with glutaraldehyde rendered the most thermostable preparation of GOX. Therefore, as the glutaraldehyde chemistry gave a high stabilization of the enzyme, we proposed another technique for improving the multipoint attachment through glutaraldehyde: the enzyme was ionically adsorbed on cationic supports with primary amino groups and then the immobilized preparation was treated with a glutaraldehyde solution. The decrease on enzyme activity was < 20%. Following this methodology, we achieved the highest stability of all the immobilization systems analyzed, showing a half-life 100 times higher than the soluble enzyme. Moreover, this derivative showed a higher stability in the presence of organic solvents (for instance methanol) or hydrogen epoxide than the ionically adsorbed enzyme or the soluble one. Therefore, the adsorption of GOX on aminated cationic support and subsequent treatment with glutaraldehyde was presented as a very successful methodology for achieving a very stable biocatalyst. (c) 2005 Elsevier B.V. All rights reserved.