Department of Chemistry , Indian Institute of Technology Bombay , Mumbai 400076 , Maharashtra India.
Department of Chemical Engineering , Indian Institute of Technology Bombay , Mumbai 400076 , Maharashtra India.
Anal Chem. 2018 Aug 7;90(15):8960-8968. doi: 10.1021/acs.analchem.8b01130. Epub 2018 Jul 26.
Phenol and its derivatives constitute a class of highly toxic xenobiotics that pollute both river and groundwater. Here, we use a highly stable enzyme-based in vitro biosensing scaffold to develop a chip-based environmental diagnostic for in situ accurate, direct detection of phenol with selectively down to 10 ppb. Mesoporous silica nanoparticles (MCM41) having a pore diameter of 6.5 nm was screened and found to be the optimal solid support for creation of a robust immobilized protein based sensor, which retains stability, enzyme activity, sensitivity, and selectivity at par with solution format. The sensor strip exhibits minimal cross reactivity in simulated wastewater, crowded with several common pollutants. Moreover, this design is competent towards detection of phenol content with 95% accuracy in real-time environmental samples collected from local surroundings, making it a viable candidate for commercialization. The enzyme has been further modified via evolution driven mutagenesis to generate an exclusive 2,3-dimethylphenol sensor with equivalent selectivity and sensitivity as the native phenol sensor. Thus, this approach can be extended to generate a battery of sensors for other priority aromatic pollutants, highlighting the versatility of the biosensor unit. This novel biosensor design presents promising potential for direct detection and can be integrated in a device format for on-site pollutant monitoring.
苯酚及其衍生物属于一类高度有毒的异生物质，会污染河流和地下水。在这里，我们使用一种高度稳定的基于酶的体外生物传感支架，开发出一种基于芯片的环境诊断方法，用于现场准确、直接检测酚，选择性低至 10 ppb。筛选出孔径为 6.5nm 的介孔硅纳米粒子（MCM41），发现其是创建稳定的固定化蛋白质传感器的最佳固体载体，该传感器在溶液形式下保留了稳定性、酶活性、灵敏度和选择性。传感器条在模拟废水中表现出最小的交叉反应性，其中充斥着几种常见的污染物。此外，该设计能够以 95%的准确度实时检测从当地环境中采集的实际环境样本中的苯酚含量，使其成为商业化的可行候选物。该酶已通过进化驱动的诱变进一步修饰，生成了一种专用于 2,3-二甲基苯酚的传感器，其选择性和灵敏度与天然苯酚传感器相当。因此，这种方法可以扩展到生成一系列用于其他优先芳香族污染物的传感器，突出了生物传感器单元的多功能性。这种新型生物传感器设计具有直接检测的巨大潜力，并可集成到设备格式中用于现场污染物监测。
