金属有机骨架材料载体用于酶固定化的研究进展

doi:10.16085/j.issn.1000-6613.2018-2451

摘要/Abstract

摘要：

现有的固定化酶技术存在低稳定性、低回收利用率等缺点，而金属有机骨架材料（MOFs）凭借自身独特的性质如特定的主-客体相互作用及限域效应，极大提高了酶的负载率以及限制酶分子的流失，甚至极端环境下仍能维持酶的活性，近年来成为固定化酶载体的研究热点。本文从酶-MOF复合物的合成策略出发，介绍了利用MOFs载体完成酶固定化包括表面吸附、共价结合、孔道包埋和原位合成这4种方法的优劣势，重点概述了酶-MOF复合物在微反应器构建、级联反应等应用领域的巨大潜力。研究进展表明，有巨大优势的新型MOFs载体将有力促进酶-MOF复合物作为优异的催化剂、生物传感器等在多学科领域的发展。

关键词: 金属有机骨架材料, 酶, 固定化, 合成策略

Abstract:

The existing immobilized enzyme technologies (mainly the ill-developed carriers) have limitations of low stability and/or low recovery rate. Metal-organic frameworks (MOFs) are considered as the most promising carriers for enzyme immobilization due to their unique properties such as specific host-guest interaction and confinement effect, which can greatly keep enzymes with high loading, low leaching and comparable catalytic efficiency under harsh conditions. In this review, four kinds of efficient synthesis strategies including surfaces adsorption, covalent binding, cage inclusion and in-situ synthesis are introduced and compared. Moreover, the potential applications of enzyme-MOF composites in micro-reactor design, cascade reaction and other fields are discussed. The research progresses indicate that the huge merits of MOFs carriers will synergistically promote the development of enzyme-MOF composites as catalyst and bio-sensor in multi-disciplinary fields.

Key words: metal-organic frameworks (MOFs), enzyme, immobilization, synthetic strategie

中图分类号:

范铮,唐咸昌,张旭,李畅,张国亮. 金属有机骨架材料载体用于酶固定化的研究进展[J]. 化工进展, 2019, 38(10): 4606-4613.

Zheng FAN,Xianchang TANG,Xu ZHANG,Chang LI,Guoliang ZHANG. Progress of on enzyme immobilization with metal-organic frameworks[J]. Chemical Industry and Engineering Progress, 2019, 38(10): 4606-4613.

图/表 6

表1 利用MOFs载体固定酶的4种主要合成策略及其比较

方法	优势	劣势
表面吸附	操作简单；绿色化学	酶与载体作用力弱，易流失；耗时；强静电相互作用可能影响蛋白质构象，从而导致酶活性下降
共价结合	酶与载体之间作用力强	操作复杂；化学试剂引入可能对酶活性有影响
孔道包埋	酶负载率高且不易流失	对MOFs孔径（大多限于介孔）、酶尺寸、形状都有要求
原位合成	制备条件温和，酶活性不受影响；快速	合成条件限于水溶液；MOFs种类较少

图1 通过疏水作用、氢键等相互作用分别将GDH和MG吸附到ZIF-70表面示意图[32]

图2 胰蛋白酶上的游离羧基与活化后MIL-88B-NH2(Cr)上的氨基发生共价结合示意图[35]

图3 自由扩散的酶分子被介孔尺寸MOFs孔道截住的示意图[23]

图4 将Cyt C和PVP共同加到2-甲基咪唑和Zn(Ⅱ)的甲醇溶液中，通过自组装生成Cyt C@ZIF-8复合物示意图[43]

表2 不同酶-MOF复合物的合成方法及其应用

方法	酶	MOF	应用	参考文献
表面吸附	微过氧化物酶（MP-11）	[Cu(OOC-C₆H₄-C₆H₄-COO)· $12$ C₆H₁₂N₂] _n	催化	[31]
表面吸附	葡萄糖脱氢酶（GDH）	ZIF-7、 ZIF-8、 ZIF-67、 ZIF-68、 ZIF-70	传感	[32]
表面吸附	洋葱伯克霍尔德氏菌脂肪酶（BCL）	An-ZIF-8	酯水解和酯交换	[33]
表面吸附	NBD标记的胰蛋白酶	MIL-100(Cr)、 MIL-101(Cr)、 UiO-66 (Zr)、 CYCU-4(Al)	蛋白质消化	[34]
共价结合	胰蛋白酶（Try）	MIL-101(Cr)、 MIL-88B(Cr)、 MIL-88B-NH₂(Cr)	蛋白质组学	[35]
共价结合	链霉亲和素（SA）	HKUST-1	DNA传感器	[36]
共价结合	β-葡萄糖苷酶	NH₂-MIL-53(A1), ZIF-67, ZIF-8	催化	[37]
孔道包埋	HRP、 Cyt C、 MP-11	PCN-332、 PCN-333	催化	[27]
孔道包埋	胰岛素、有机磷酸酐酶（OPAA）	NU-100x、 PCNs	解毒	[28,30,40]
孔道包埋	葡萄糖氧化酶（GOx）, HRP	PCN-888	催化	[57]
原位合成	卵清蛋白、核糖核酸酶A、人血清白蛋白、吡咯喹啉醌依赖性葡萄糖脱氢酶、脂肪酶、血红蛋白、溶菌酶、胰岛素、 HRP、 Try、脲酶和寡核苷酸	ZIF-8、 HKUST-1、 Eu/Tb-BDC、 MIL-88A	生物样本库、生物反应器	[29,44]
原位合成	Cyt C、 HRP、脂肪酶（经PVP修饰）	ZIF-8、 ZIF-10	催化，检测	[43]
原位合成	GOx, HRP	ZIF-8	催化，检测	[54]

表2 不同酶-MOF复合物的合成方法及其应用

方法	酶	MOF	应用	参考文献
表面吸附	微过氧化物酶（MP-11）	[Cu(OOC-C₆H₄-C₆H₄-COO)· $12$ C₆H₁₂N₂] _n	催化	[31]
表面吸附	葡萄糖脱氢酶（GDH）	ZIF-7、 ZIF-8、 ZIF-67、 ZIF-68、 ZIF-70	传感	[32]
表面吸附	洋葱伯克霍尔德氏菌脂肪酶（BCL）	An-ZIF-8	酯水解和酯交换	[33]
表面吸附	NBD标记的胰蛋白酶	MIL-100(Cr)、 MIL-101(Cr)、 UiO-66 (Zr)、 CYCU-4(Al)	蛋白质消化	[34]
共价结合	胰蛋白酶（Try）	MIL-101(Cr)、 MIL-88B(Cr)、 MIL-88B-NH₂(Cr)	蛋白质组学	[35]
共价结合	链霉亲和素（SA）	HKUST-1	DNA传感器	[36]
共价结合	β-葡萄糖苷酶	NH₂-MIL-53(A1), ZIF-67, ZIF-8	催化	[37]
孔道包埋	HRP、 Cyt C、 MP-11	PCN-332、 PCN-333	催化	[27]
孔道包埋	胰岛素、有机磷酸酐酶（OPAA）	NU-100x、 PCNs	解毒	[28,30,40]
孔道包埋	葡萄糖氧化酶（GOx）, HRP	PCN-888	催化	[57]
原位合成	卵清蛋白、核糖核酸酶A、人血清白蛋白、吡咯喹啉醌依赖性葡萄糖脱氢酶、脂肪酶、血红蛋白、溶菌酶、胰岛素、 HRP、 Try、脲酶和寡核苷酸	ZIF-8、 HKUST-1、 Eu/Tb-BDC、 MIL-88A	生物样本库、生物反应器	[29,44]
原位合成	Cyt C、 HRP、脂肪酶（经PVP修饰）	ZIF-8、 ZIF-10	催化，检测	[43]
原位合成	GOx, HRP	ZIF-8	催化，检测	[54]

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