Lekbach Yassir, Ueki Toshiyuki, Liu Xiaomeng, Woodard Trevor, Yao Jun, Lovley Derek R
Department of Microbiology, University of Massachusetts, Amherst, MA, 01003, USA.
Department of Electrical and Computer Engineering University of Massachusetts, Amherst, MA, 01003, USA.
Biosens Bioelectron. 2023 Apr 15;226:115147. doi: 10.1016/j.bios.2023.115147. Epub 2023 Feb 10.
Nanowires have substantial potential as the sensor component in electronic sensing devices. However, surface functionalization of traditional nanowire and nanotube materials with short peptides that increase sensor selectivity and sensitivity requires complex chemistries with toxic reagents. In contrast, microorganisms can assemble pilin monomers into protein nanowires with intrinsic conductivity from renewable feedstocks, yielding an electronic material that is robust and stable in applications, but also biodegradable. Here we report that the sensitivity and selectivity of protein nanowire-based sensors can be modified with a simple plug and play genetic approach in which a short peptide sequence, designed to bind the analyte of interest, is incorporated into the pilin protein that is microbially assembled into nanowires. We employed a scalable Escherichia coli chassis to fabricate protein nanowires that displayed either a peptide previously demonstrated to effectively bind ammonia, or a peptide known to bind acetic acid. Sensors comprised of thin films of the nanowires amended with the ammonia-specific peptide had a ca. 100-fold greater response to ammonia than sensors made with unmodified protein nanowires. Protein nanowires with the peptide that binds acetic acid yielded a 4-fold higher response than nanowires without the peptide. The protein nanowire-based sensors had greater responses than previously reported sensors fabricated with other nanomaterials. The results demonstrate that protein nanowires with enhanced sensor response for analytes of interest can be fabricated with a flexible genetic strategy that sustainably eliminates the energy, environmental, and health concerns associated with other common nanomaterials.
纳米线作为电子传感设备中的传感器组件具有巨大潜力。然而,用短肽对传统纳米线和纳米管材料进行表面功能化以提高传感器的选择性和灵敏度,需要使用有毒试剂的复杂化学方法。相比之下,微生物可以将菌毛蛋白单体组装成具有本征导电性的蛋白质纳米线,其原料可再生,所产生的电子材料在应用中坚固稳定,而且可生物降解。在此,我们报告基于蛋白质纳米线的传感器的灵敏度和选择性可以通过一种简单的即插即用基因方法进行修饰,即在微生物组装成纳米线的菌毛蛋白中掺入一个设计用于结合目标分析物的短肽序列。我们采用了一种可扩展的大肠杆菌底盘来制造蛋白质纳米线,这些纳米线展示了一种先前已证明能有效结合氨的肽,或者一种已知能结合乙酸的肽。由用氨特异性肽修饰的纳米线薄膜组成的传感器对氨的响应比用未修饰蛋白质纳米线制成的传感器高约100倍。带有结合乙酸肽的蛋白质纳米线比没有该肽的纳米线产生的响应高4倍。基于蛋白质纳米线的传感器比先前报道的用其他纳米材料制造的传感器具有更高的响应。结果表明,通过一种灵活的基因策略可以制造出对目标分析物具有增强传感器响应的蛋白质纳米线,该策略可持续地消除了与其他常见纳米材料相关的能源、环境和健康问题。