Chen Haiying, Huang Baojian, Han Lei
College of Chemistry and Pharmaceutical Sciences, Qingdao Agricultural University, 700 Changcheng Road, Qingdao, Shandong 266109, China.
College of Chemistry and Pharmaceutical Sciences, Qingdao Agricultural University, 700 Changcheng Road, Qingdao, Shandong 266109, China.
J Hazard Mater. 2025 Jul 5;491:137957. doi: 10.1016/j.jhazmat.2025.137957. Epub 2025 Mar 17.
Designing a portable device based on bacterial laccase (BLac) for on-site assay of phenolic contaminants presents significant challenges. Here, we achieved comprehensive performance enhancement of BLac by integrating biomineralization and microbial surface display technologies. The introduction of cell surface-displayed bacterial laccase (CSD-BLac) reduced costs and improved sensitivity compared to detection methods based on free Blac and whole-cell catalyst. Further, the biomineralization dramatically enhanced the catalytic efficiency (V/K) of mineralized CSD-BLac (M-CSD-BLac), making it 1.98 times higher than that of CSD-BLac. Mineralization conditions could significantly affect the activity of M-CSD-Blac. Moreover, the biomineralization layer also enhanced the resistance of M-CSD-BLac against high temperature, metal ions, ionic strength and storage time. Further, a portable assay device was developed for detection of phenolic pollutants by depositing M-CSD-BLac on a syringe filter membrane, which demonstrated easy operation, rapid detection (10 min), good reusability (20 cycles). The device not only could reliably differentiate three types of phenols but also quantitatively detect them with high sensitivity. For phenol, m-aminophenol, and p-nitrophenol, the limits of detection were 0.09, 0.28 and 0.17 μM, with detection ranges of 10-70, 20-80 and 15-110 μM, respectively. Additionally, the porous structure of M-CSD-BLac layer and the insertion of M-CSD-BLac into the filter membrane pores allowed effective filtration of smaller pigments from real samples, eliminating the need for additional pretreatment. This work not only proposes a strategy for elevating the activity and stability of laccase, but also stimulates the development of portable assay devices for on-site environmental monitoring.
设计一种基于细菌漆酶（BLac）的便携式设备用于现场检测酚类污染物面临重大挑战。在此，我们通过整合生物矿化和微生物表面展示技术实现了BLac综合性能的提升。与基于游离BLac和全细胞催化剂的检测方法相比，细胞表面展示细菌漆酶（CSD - BLac）的引入降低了成本并提高了灵敏度。此外，生物矿化显著提高了矿化CSD - BLac（M - CSD - BLac）的催化效率（V/K），使其比CSD - BLac高1.98倍。矿化条件会显著影响M - CSD - Blac的活性。此外，生物矿化层还增强了M - CSD - BLac对高温、金属离子、离子强度和储存时间的耐受性。进一步地，通过将M - CSD - BLac沉积在注射器滤膜上开发了一种用于检测酚类污染物的便携式检测设备，该设备操作简便、检测快速（10分钟）、具有良好的可重复使用性（20个循环）。该设备不仅能够可靠地区分三种酚类，还能以高灵敏度对它们进行定量检测。对于苯酚、间氨基酚和对硝基苯酚，检测限分别为0.09、0.28和0.17μM，检测范围分别为10 - 70、20 - 80和15 - 110μM。此外，M - CSD - BLac层的多孔结构以及M - CSD - BLac插入滤膜孔中能够有效过滤实际样品中较小的色素，无需额外的预处理。这项工作不仅提出了提高漆酶活性和稳定性的策略，还推动了用于现场环境监测的便携式检测设备的发展。
