合成生物纳米酶

doi:10.12211/2096-8280.2022-009

摘要/Abstract

摘要：

纳米酶是一类本身蕴含酶学特性的纳米材料，能够在生理条件下催化天然酶的底物及其介导的生化反应，表现出类似的反应动力学和催化机理。自2007年首次发现四氧化三铁纳米颗粒自身具有类过氧化物酶活性以来，成百上千种不同的纳米酶材料相继被开发出来，在生物医学、检测传感、环境工程等领域有着广泛的应用。近年来，以基因编辑或重组为基础的纳米酶材料开始出现。相比较其他纳米酶，这种纳米酶是基于合成生物学相关技术开发而来，在这里将其命名为合成生物纳米酶，其特点是以人为改造或从头设计的蛋白作为骨架，原位生长一些金属纳米颗粒，将蛋白骨架的功能和材料的催化融于一体。本文主要介绍了纳米酶的基本概况，例证了其在生物医学应用上的优势；概述了多种天然蛋白作为骨架制备纳米酶的原理，列举了其中的部分应用；简述了基因改造蛋白骨架方面的研究进展，并重点强调这种蛋白骨架在合成无机纳米颗粒方面的优势；在以上这些进展的基础上，提出了合成生物纳米酶的概念，并阐释了其中的内涵，最后也以基因重组铁蛋白纳米酶为例介绍了目前的一些设计及应用。未来，以合成生物纳米酶为代表的纳米酶，有可能会将计算生物学、结构生物学、蛋白/基因工程及化学等手段融为一体，在模拟酶的设计上更为理性，在赋予功能上更为多样化，并且有望进一步促进合成生物学与纳米生物学的深度融合。

关键词: 纳米酶, 纳米生物材料, 基因改造, 合成生物纳米酶, 铁蛋白纳米酶

Abstract:

Nanozymes are nanomaterials with intrinsic enzyme-like activities, which can convert specific substrates to products and catalyze biochemical reactions as natural enzymes, with the similar reaction dynamics and catalytic mechanisms. Since the discovery of the intrinsic peroxidase activity of Fe₃O₄ nanoparticles, thousands of nanozymes have been developed, and extensively applied in many fields, such as biomedicine, biosensor/biodetection and bioremediation. Recently, one type of nanozymes based on gene editing or genetic recombination technology was developed. Compared to other nanozymes, this kind of nanozymes was fabricated based on the key techniques of synthetic biology. Herein, we term them as synthetic biological nanozymes. The main characteristic of synthetic biological nanozymes is the utilization of artificially or de novo designed protein as scaffolds. Furthermore, the amino acids or domains of the scaffolds with metal ion binding ability can be used for synthesizing metal nanoparticles. This characteristic of synthetic biological nanozymes integrates the function of proteins and the catalytic activity of metal nanoparticles together. In this review, we comment the progress of nanozymes and their advantages in biomedicine applications, and highlight the synthesis principle of natural protein scaffold-based nanozymes and their applications in nanomedicine. We also introduce the gene editing or genetic recombination protein scaffolds for nanomaterials preparation and the superiorities of this kind of scaffolds for inorganic nanoparticles synthesis. Through such an overview, we elaborate the definition of synthetic biological nanozymes, and explain their essential connotation and finally take ferritin-based nanozymes as an example to demonstrate their design and applications. In the future, synthetic biological nanozymes may integrate technological innovations of many fields including computational biology, structural biology, protein engineering, genetic engineering, chemistry, etc., which would make the design of nanozymes more rational, their functions more diverse, and integration of synthetic biology and nanobiology more profound.

Key words: nanozymes, bioactive materials, genetic engineering, synthetic biological nanozymes, ferritin nanozymes

中图分类号:

Q816

刘奇奇, 王春玉, 齐天翊, 朱明盛, 黄兴禄. 合成生物纳米酶[J]. 合成生物学, 2022, 3(2): 320-334.

Qiqi LIU, Chunyu WANG, Tianyi QI, Mingsheng ZHU, Xinglu HUANG. Synthetic biological nanozyme[J]. Synthetic Biology Journal, 2022, 3(2): 320-334.

图/表 7

表1 天然酶与纳米酶优缺点对比［9］

Tab. 1 Advantages and disadvantages of natural enzymes and nanozymes［9］

纳米酶		天然酶
优点	缺点	优点	缺点
高催化活性、成本低	活性类型有限	高催化活性	成本高
稳定性高	底物选择性有限	高底物选择性	稳定性有限
可长期存储	具有潜在的纳米毒性	良好的生物相容性	长期存储难
易于批量生产、可回收利用	催化机制不清楚	生物催化类型广泛	批量生产难、分离纯化耗时
多酶模拟活性、可控催化活性和类型	缺乏统一标准和参考资料	应用广泛	催化活性单一
易于多功能化（生物共轭表面积大）、可以通过外部刺激进行催化活性和催化类型控制（如光、超声、热、磁等）	催化性能依赖于大小、形状、结构和成分等	通过基因和蛋白质工程进行理性设计
适用于极端环境			在极端环境中使用难（如高温、极端pH、盐离子浓度、紫外线照射等）
独特的物理化学性质（如荧光、电、顺磁性质等）

图1 纳米酶试纸条的设计［24］（a）标准胶体金试纸条；（b）纳米酶试纸条，利用纳米酶的活性，可以与底物发生显色反应，实现肉眼可见的增强信号

Fig. 1 Design for nanozyme-strips［24］(a) Colloidal gold strips; (b) Nanozyme-strips. Probe with nanozyme activity generates a color reaction with substrates, which significantly enhances the signal so that it can be visualized by naked-eyes

图2 HSA的晶体结构［40］

Fig. 2 Crystal structure of HSA［40］

图3 各种基于笼状蛋白的结构示意图及其透射电镜图［52］（比例尺= 50 nm。上图显示了基于天然结构的纳米笼，下图展示了基于病毒样颗粒的纳米笼）

Fig. 3 Schematic diagrams and TEM images of structures of various protein-based nanocages[52](Scale bar: 50 nm. Upper images show natural structure-based nanocages, and lower images show virus-like particles.)

图4 杂化铁蛋白纳米酶（mito-fenozyme）保护线粒体功能［88］（a）mito-fenozyme制备示意图；（b）FTn的负染色TEM图；（c）MnO2 fenozyme TEM图；（d） mito-fenozyme TEM图（比例尺=20 nm）；（e）在mito-fenozyme作用下细胞内自由基向非细胞毒性分子转化的示意图；（f）mito-fenozyme影响线粒体氧化损伤的共聚焦图像（上）和量化分析（下）；（g） mito-fenozyme处理的细胞内氧化损伤自由基水平下降（比例尺=10 μm）

Fig. 4 Hybrid ferritin nanozyme (Mito-Fenozyme) protects mitochondrial functions［88］(a) Schematic diagram for the preparation of Mito-Fenozyme. TEM images of the FTn protein shell (negative staining with 1% uranyl acetate) (b), MnO2 nanoparticles inside the FTn shell (c), and Mito-Fenozyme (d). Scale bar: 20 nm. (e) Schematic diagram for the intracellular conversion of free radicals to non-cytotoxic molecules under the catalysis of Mito-Fenozyme. (f) Confocal images (top) and quantitative analysis (bottom) for the effect of Mito-Fenozyme on mitochondrial oxidative damage. (g) Mito-Fenozyme reduced intracellular free radical levels in oxidatively damaged cells. Scale bar: 10 μm

图5 模块化组装思路构筑寡聚纳米酶［87］（a）铁蛋白胞内重组胞外再重组构筑寡聚组装体示意图；（b）构筑的几种组装体的示意图（上）和TEM照片（下）。比例尺=20 nm

Fig. 5 Assembly of chain-like nanostructures［87］(a) Schematic illustration for the preparation of FTn-Ner via a two-step self-assembly/post-assembly approach. The uniform FTn as motifs were obtained by the self-assembly of 24 FTn subunits expressed in E. coli. Subsequently, purified FTn motifs were further assembled to form different FTn-Ner by using two-armed PEG. (b) Representative TEM images of various assembled nanostructures. Scale bar: 20 nm

图6 合成生物纳米酶可能的方向

Fig. 6 Prospects for synthetic biological nanozymes

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