Cohesive Living Bacterial Films with Tunable Mechanical Properties from Cell Surface Protein Display.

IF 3.7 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS ACS Synthetic Biology Pub Date : 2024-11-01 DOI:10.1021/acssynbio.4c00528
Hanwei Liu, Priya K Chittur, Julia A Kornfield, David A Tirrell
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Abstract

Engineered living materials (ELMs) constitute a novel class of functional materials that contain living organisms. The mechanical properties of many such systems are dominated by the polymeric matrices used to encapsulate the cellular components of the material, making it hard to tune the mechanical behavior through genetic manipulation. To address this issue, we have developed living materials in which mechanical properties are controlled by the cell-surface display of engineered proteins. Here, we show that engineered Esherichia coli cells outfitted with surface-displayed elastin-like proteins (ELPs, designated E6) grow into soft, cohesive bacterial films with biaxial moduli around 14 kPa. When subjected to bulge-testing, such films yielded at strains of approximately 10%. Introduction of a single cysteine residue near the exposed N-terminus of the ELP (to afford a protein designated CE6) increases the film modulus 3-fold to 44 kPa and eliminates the yielding behavior. When subjected to oscillatory stress, films prepared from E. coli strains bearing CE6 exhibit modest hysteresis and full strain recovery; in E6 films much more significant hysteresis and substantial plastic deformation are observed. CE6 films heal autonomously after damage, with the biaxial modulus fully restored after a few hours. This work establishes an approach to living materials with genetically programmable mechanical properties and a capacity for self-healing. Such materials may find application in biomanufacturing, biosensing, and bioremediation.

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通过细胞表面蛋白质展示具有可调机械特性的粘性活细菌薄膜
工程活体材料(ELMs)是一类含有活生物体的新型功能材料。许多此类系统的机械特性受用于封装材料细胞成分的聚合物基质的支配,因此很难通过基因操纵来调整机械行为。为了解决这个问题,我们开发出了由细胞表面显示的工程蛋白质控制机械特性的活体材料。在这里,我们展示了装有表面显示弹性蛋白(ELPs,命名为 E6)的工程大肠杆菌细胞生长成柔软、内聚的细菌薄膜,其双轴模量约为 14 kPa。在进行隆起测试时,这种薄膜的应变约为 10%。在 ELP 暴露的 N 端附近引入一个半胱氨酸残基(产生一种名为 CE6 的蛋白质),可将薄膜模量提高 3 倍,达到 44 千帕,并消除屈服行为。当受到振荡应力时,由含有 CE6 的大肠杆菌菌株制备的薄膜会表现出适度的滞后和完全的应变恢复;而在 E6 薄膜中则会观察到更明显的滞后和大量的塑性变形。CE6 薄膜在受损后可自主愈合,双轴模量在几小时后完全恢复。这项研究为具有基因可编程机械特性和自愈合能力的活体材料提供了一种方法。这种材料可应用于生物制造、生物传感和生物修复领域。
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来源期刊
CiteScore
8.00
自引率
10.60%
发文量
380
审稿时长
6-12 weeks
期刊介绍: The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.
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Cohesive Living Bacterial Films with Tunable Mechanical Properties from Cell Surface Protein Display. A Plug-and-Play T7 Expression System for Heterologous Production of Lanthipeptides in Bacillus subtilis. Design of a Genetically Programmable and Customizable Protein Scaffolding System for the Hierarchical Assembly of Robust, Functional Macroscale Materials. Recent Advances in Genetic Engineering Strategies of Sinorhizobium meliloti. Using Cupriavidus necator H16 to Provide a Roadmap for Increasing Electroporation Efficiency in Nonmodel Bacteria.
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