One-pot synthesis of trypsin-based magnetic metal-organic frameworks for highly efficient proteolysis.
One-pot synthesis of trypsin-based magnetic metal-organic frameworks for highly efficient proteolysis.
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DOI：
10.1039/c9tb02315a
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发表时间：
2020-05
期刊：
影响因子：
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通讯作者：
Chao Zhong;Zhixian Lei;Huan Huang;Mingyue Zhang;Z. Cai;Zian Lin
中科院分区：
文献类型：
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作者：
Chao Zhong;Zhixian Lei;Huan Huang;Mingyue Zhang;Z. Cai;Zian Lin
通过仿生矿化方法将酶固定化到金属有机骨架(MOF)上，可以在恶劣的环境中保存生物功能。尽管这种方法取得了成功，但由于MOF的有机单体所造成的反应体系的碱性环境，使得它不适合一些pH敏感的酶，特别是对胰酶。本文报道了一种简便的一锅法合成固定化胰酶磁性分子筛咪唑骨架-8(氧化铁@ZIF-8@胰酶)的方法，当反应体系的pH变化到一定程度时，ZIF-8在柠檬酸修饰的氧化铁周围生长和胰酶的固定化同时发生。氧化铁具有较大的比表面积和较高的载酶能力，其酶活性是游离胰酶的2.6倍。此外，它还表现出良好的磁响应性(43emu g-1)，使操作和回收变得容易和方便。此外，氧化铁@ZIF-8@胰酶可以作为固定化酶微反应器(Imer)，快速高效地消化蛋白质和复杂的人血清样品，结果令人满意，在蛋白质组学分析中具有广阔的应用前景。
Immobilization of enzymes onto metal-organic frameworks (MOFs) through a biomimetic mineralization approach can preserve biological functionality in harsh environments. Despite the success of this approach, the alkaline environment of the reaction system, which is caused by the organic monomers of MOFs, makes it unsuitable for some pH-sensitive enzymes, especially for trypsin. Herein, we reported a facile approach for the one-pot synthesis of trypsin-immobilized magnetic zeolite imidazolate framework-8 (iron oxide@ZIF-8@trypsin), where the growth of ZIF-8 around the citric acid-modified iron oxide and immobilization of trypsin occurred simultaneously when the pH of the reaction system was changed to some extent. With a large specific surface area and a high enzyme loading capacity, the resultant iron oxide@ZIF-8@trypsin exhibited 2.6 times higher enzymatic activity than free trypsin. Moreover, it showed a favourable magnetic response (43 emu g-1) which made the operation and recycling easy and convenient. In addition, iron oxide@ZIF-8@trypsin could be applied as an immobilized enzyme microreactor (IMER) to rapidly and efficiently digest proteins and complex human serum samples with satisfactory results, showing great promise for application in proteomic analysis.