生物合成高性能蛋白及材料应用

doi:10.12211/2096-8280.2022-032

摘要/Abstract

摘要：

源自高性能生物结构蛋白的新一代材料体系在高技术领域具有广阔的应用前景。随着基因组装、定点突变和合成改造等合成生物学工具的发展，具有新特性的人工高性能蛋白及其多级材料体系获得了极大拓展。然而，目前在人工蛋白的高效生物合成、理性功能设计和跨尺度体外组装等方面仍面临严峻挑战。本文综述了利用合成生物学方法设计合成高性能蛋白材料的研究进展，强调了合成生物学技术在人工蛋白定向优化、结构改造和可编程材料组装等方面的作用，并突出了高性能蛋白及组装体在构建高强蛋白纤维和黏合材料领域的应用。最后，面向高性能蛋白材料理性设计和规模化制备的重大需求，对具有发展潜力的新型蛋白分子和技术平台进行了总结和展望，为今后该领域的应用基础研究提供了可借鉴的思路。

关键词: 生物材料, 合成生物学, 蛋白工程, 纤维, 黏合剂

Abstract:

Biomaterials, derived from high-performance structural proteins such as elastin, spidroin, and resilin, exhibit broad applications in high-tech fields from wearable devices, biomedicine to military scenarios. Advanced synthetic biology tools including genetic recombination, site-directed mutagenesis, and metabolic pathway optimization enable the generation of recombinant structural proteins with customized properties. Additionally, such recombinant proteins could further assemble into disordered aggregates or ordered hierarchies to fulfill multiple functionalities. In particular, two or more independently natural structure proteins have been connected to produce recombinant proteins as scaffolds with superior mechanical properties. The mechanical performance has surpassed chemically synthesized polymers. By harnessing the power of metabolic pathway optimization, engineered microbes could enable the large production of high-molecular-weight proteins as renewable materials. Furthermore, through the integration of synthetic biology and materials science, programmable materials with multiple composition and sophisticated spatial organization have been fabricated, with diverse functionalities including self-repair, information storage, impact resistance, wound healing, etc. The protein-based materials established a new niche for current material systems. Despite recent advances, there are still huge challenges to be addressed in rational design, scalable production, and systematic integration for recombinant proteins. Synthetic biology, coupling with advanced techniques in materials science, provides a feasible route to tackle the aforementioned challenges and promote the innovation in bioinspired protein materials. In this review, we outline key advances in the design and fabrication of biosynthetic protein-based materials. First, we highlight the achievements in protein design, structural reconstruction and programmable assembly of novel biomaterials. Next, we discuss the developments of typical advanced protein materials, including biofibers and bioadhesives. Finally, we envision the ideal biosynthetic platforms, which would enable the rational de novo design and mass production for protein-based materials in the future.

Key words: biomaterials, synthetic biology, protein engineering, fibers, adhesives

中图分类号:

Q816

李敬敬, 马超, 王帆, 张洪杰, 刘凯. 生物合成高性能蛋白及材料应用[J]. 合成生物学, 2022, 3(4): 638-657.

Jingjing LI, Chao MA, Fan WANG, Hongjie ZHANG, Kai LIU. Biosynthesis of high-performance protein materials and their applications[J]. Synthetic Biology Journal, 2022, 3(4): 638-657.

图/表 7

图1 基于多生物融合的人工结构蛋白的设计（a）、模块重组（b）与生物合成（c）

Fig. 1 Schematic diagram of design (a), recombination (b) and biosynthesis (c) of the man-made structural proteins

图2 动态共价相互作用介导的人工结构蛋白组装

Fig. 2 Dynamic covalent interactions-mediated assembly of the recombinant structural proteins

图3 非共价相互作用介导的人工结构蛋白组装（a）蛋白末端结构域及折叠结构内部作用诱导的蛋白有序自组装［71］；（b）人工蛋白与长链DNA分子的复合组装［61］；（c～d）超电荷重组类弹性蛋白与表面活性剂分子的复合组装［72-73］

Fig. 3 Non-covalent interactions-mediated assembly of the recombinant structural proteins(a) Protein self-assembly induced by both C-terminal domain and internal actions among secondary structural elements[71]; (b) Composite assembly of recombinant structural proteins and DNA molecules[61]; (c～d) Composite assembly of supercharged recombinant structural proteins and surfactant molecules[72-73]

表1 高性能蛋白纤维和黏合剂材料性能汇总

Tab. 1 Properties of high-performance protein fibers and adhesives

序号	蛋白	氨基酸序列	修饰方法	材料性能	参考文献
1	蓑衣虫丝蛋白	(A) _n -(GA) _n -Linker-(GAGAGS) _n	天然蛋白	拉伸强度2.0 GPa，韧性364 MJ/m³	[22]
2	重组鱿鱼环齿蛋白	(PAATAVSHTTHHAP) [VPGVG(VPGKG) _n ]	不同折叠结构的基序重组	拉伸强度650 MPa，韧性120 MJ/m³	[34]
3	重组蛛丝蛋白	[(GGX) _n FGAILSS]₁₂₈	不同折叠结构的基序重组与多聚化	拉伸强度0.98 GPa，韧性161 MJ/m³	[35]
4	重组蛛丝蛋白	[An(GGX) _n ]₁₉₂	蛋白共价组装	拉伸强度1.03 GPa，韧性114 MJ/m³	[59]
5	重组节肢弹性蛋白	(VGSGGRPSDSYGAPGGGNP) [VPGVG(VPGKG) _n ]	蛋白-海藻酸盐复合	拉伸强度768 MPa，模量24 MPa，韧性52 MJ/m³	[63]
5	重组鱿鱼环齿蛋白	(PAATAVSHTTHHAP) [VPGVG(VPGKG) _n ]	蛋白-海藻酸盐复合	拉伸强度606 MPa，韧性69 MJ/m³	[63]
6	重组蛛丝蛋白	NTD-[A _n (GGX) _n ]-CTD	蛋白非共价组装	拉伸强度834 MPa，韧性143 MJ/m³	[71]
7	重组节肢弹性蛋白	(VGSGGRPSDSYGAPGGGNP) [VPGVG(VPGKG) _n ]	蛋白-DNA复合	韧性250 MJ/m³	[61]
8	重组类弹性蛋白	[(VPGVG)₂₀pAzF]₄	非天然氨基酸定点修饰	光交联反应活性	[27]
9	重组贻贝足丝蛋白	Mfp5-CsgA	不同生物功能的基序进行重组	水下黏合性能20.9 mJ/m²	[31]
10	重组类弹性蛋白	[VPGVG(VPGKG) _n ]	蛋白-表面活性剂复合	黏合性能16.5 MPa	[72]
11	重组类弹性蛋白	[VPGVG(VPGKG) _n ]	蛋白-表面活性剂-金属离子复合	黏合性能16 MPa	[73]
12	重组类弹性蛋白	[VPGVG(VPGKG) _n ]	蛋白-表面活性剂-Au复合	黏合性能20 MPa	[88]

图4 高性能蛋白纤维材料在生物体内的应用

Fig. 4 Applications of high-performance protein fibers in vivo

图5 高性能蛋白纤维材料在抗冲击（a）［62］和信息存储领域（b）［46］的应用

Fig. 5 Applications of high-performance protein fibers for impact resistance (a)[62] and information storage (b)[46]

图6 高性能蛋白黏合材料在生物体内的应用（a～b）超电荷重组类弹性蛋白-表面活性剂分子蛋白黏合剂在止血和伤口黏合中的应用［72-73］；（c）重组类弹性蛋白双网络静电复合蛋白黏合体系及软骨修复应用［115］；（d）近红外光响应的蛋白纳米黏合体系及肿瘤光热治疗［88］

Fig. 6 Applications of high-performance protein adhesives in vivo(a～b) Supercharged ELPs-based adhesive and the applications in hemostasis and wound repair[72-73]; (c) Fabrication of the engineered protein adhesive as a conjoined network and applications in cartilage repair[115]; (d) Fabrication of protein photothermal bioplaster (protein-SDBS-GNRs) and non-invasive skin tumor therapy[88]

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