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碳包覆金属纳米磁体上共价连接酶的高效磁回收

Efficient magnetic recycling of covalently attached enzymes on carbon-coated metallic nanomagnets.

作者信息

Zlateski Vladimir, Fuhrer Roland, Koehler Fabian M, Wharry Scott, Zeltner Martin, Stark Wendelin J, Moody Thomas S, Grass Robert N

机构信息

Institute for Chemical and Bioengineering, ETH Zurich , 8093 Zurich, Switzerland.

出版信息

Bioconjug Chem. 2014 Apr 16;25(4):677-84. doi: 10.1021/bc400476y. Epub 2014 Apr 4.

Abstract

In the pursuit of robust and reusable biocatalysts for industrial synthetic chemistry, nanobiotechnology is currently taking a significant part. Recently, enzymes have been immobilized on different nanoscaffold supports. Carbon coated metallic nanoparticles were found to be a practically useful support for enzyme immobilization due to their large surface area, high magnetic saturation, and manipulatable surface chemistry. In this study carbon coated cobalt nanoparticles were chemically functionalized (diazonium chemistry), activated for bioconjugation (N,N-disuccinimidyl carbonate), and subsequently used in enzyme immobilization. Three enzymes, β-glucosidase, α-chymotrypsin, and lipase B were successfully covalently immobilized on the magnetic nonsupport. The enzyme-particle conjugates formed retained their activity and stability after immobilization and were efficiently recycled from milliliter to liter scales in short recycle times.

摘要

在为工业合成化学寻求强大且可重复使用的生物催化剂的过程中,纳米生物技术目前正发挥着重要作用。最近,酶已被固定在不同的纳米支架载体上。由于其大表面积、高磁饱和度和可操控的表面化学性质,碳包覆金属纳米颗粒被发现是一种实际有用的酶固定化载体。在本研究中,碳包覆钴纳米颗粒经过化学功能化(重氮化学)、生物共轭活化(N,N - 二琥珀酰亚胺基碳酸酯),随后用于酶固定化。三种酶,β - 葡萄糖苷酶、α - 胰凝乳蛋白酶和脂肪酶B成功地共价固定在磁性载体上。固定化后形成的酶 - 颗粒共轭物保留了它们的活性和稳定性,并且在短循环时间内能够以毫升到升的规模高效回收利用。

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