基于活细胞的微生物砷离子传感器的性能拓展与优化  

Performance Expansion and Optimization of Live Cell-based Arsenic Ion Microbial Sensors

作　　者：崔明航 李佳佳 汪劲[1,2,4] 汪尔康 房晓娜[3] CUI Ming-Hang;LI Jia-Jia;WANG Jin;WANG Er-Kang;FANG Xiao-Na(State Key Laboratory of Electroanalytical Chemistry,Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun 130022,China;School of Applied Chemistry and Engineering,University of Science and Technology of China,Hefei 230026,China;School of Chemistry,Northeast Normal University,Changchun 130024,China;Department of Chemistry,The State University of New York at Stony Brook,New York 11794,United States of America)

机构地区：[1]中国科学院长春应用化学研究所,电分析化学国家重点实验室,长春130022 [2]中国科学技术大学应用化学与工程学院,合肥230026 [3]东北师范大学化学学院,长春130024 [4]纽约州立大学石溪分校化学系,纽约11794

出　　处：《分析化学》2024年第6期885-892,共8页Chinese Journal of Analytical Chemistry

基　　金：中央高校基本科研业务费项目(No.2412022QD011);国家自然科学基金项目(Nos.32171245,21721003)资助。

摘　　要：在全细胞微生物传感器的开发中,细胞的代谢活性状态和信号输出形式等因素会直接影响传感器的稳定性和可重复性等性能,给传感器的应用开发中标准化方法的建立带来了挑战。本研究选择基于Replication protein L(RepL)信号放大器的砷离子传感器为研究对象,探究了不同条件对砷离子传感器的检测性能的影响。研究结果表明,细胞的培养环境、生长状态(如不同的生长期)、报告子的类型以及诱导时长等均会对砷离子传感器的检测性能产生显著影响。传感器在不同培养基中的稳定性差别较大,其中在LB培养基中展现出更高的稳定性。同时,不同生长期的细胞也展现出不同的检测性能优势:在平台生长期的细胞中,传感器的检测灵敏度和线性度更佳;在对数期的细胞中,检出限更低。此外,传感器的响应存在最优的诱导时长,过长或过短的诱导时间都会影响传感器的响应,本研究中砷离子传感器的最佳诱导时间约为2~3 h。通过对3种荧光蛋白报告子的对比研究发现,荧光蛋白的选取对传感器检出限的影响不大(均在5~10μg/L范围内),但会显著改变传感器的响应时间(从40 min到2 h);同时,荧光蛋白的发光亮度越高,传感器响应越快。本研究结果表明,可以根据不同的需求选取不同状态的细胞,以最大限度优化细胞传感器的检测性能,拓展传感器的应用空间。本研究结果为微生物传感器在实际检测中的应用提供了可靠的依据。During the development of whole-cell microbial sensors,factors such as cellular metabolic activity and signal output modes play pivotal roles in the stability and repeatability of the sensors,presenting numerous challenges for the standardization of sensor applications.This research focused on the arsenic ion sensor based on the RepL amplifier,adjusting the reporter genes,culture media,growth stages,and induction times of arsenic ions,aiming to investigate how these factors affect the sensor′s detection performance.The resultsindicated that the cell′s culturing environment,growth status(e.g.,different growth phases),type of reporter,and induction time all had significant impacts on the performance of the arsenic ion sensor.First,the stability of the sensors varied greatly in different media,all the three sensors displayed greater stability in LB culture medium.Meanwhile,the cells in different growth stages also exhibited different performance advantages.Cells at the stationary growth phase exhibited better detection sensitivity and linearity,while cells in the logarithmic growth phase had lower limit of detection(LOD).Moreover,there was an optimal induction time for the response of the sensor,overly long or short induction time could interfere with its response.The optimal induction time for the arsenic sensor in this work was about 2–3 h.By comparing three types of fluorescent protein reporters,it wasfound that although their detection limits were fairly similar,all within the range of 5–10μg/L,but their response times varied,ranging from 40 min to 2 h.The fluorescent proteins with higher brightness exhibited faster sensor response.These research outcomes provided a solid foundation for the practical application of microbes in detection.In practice,we could choose cells in specific states based on particular purpose,maximizing the performance of the cell sensors and further broadening the application scope of such sensors.

关 键 词：砷离子微生物传感器 培养基 细菌生长阶段 荧光蛋白报告基因 响应时间 

分 类 号：Q813.11[生物学—生物工程] O657.3[理学—分析化学] TP212[理学—化学]