title: Construction of 2,4,6-Trinitrotoluene Biosensors with Novel Sensing Elements from Escherichia coli K-12 MG1655.
abstract: Detection of 2,4,6-trinitrotoluene (TNT) has been extensively studied since it is a common explosive filling for landmines, posing significant threats to the environment and human safety. The rapid advances in synthetic biology give new hope to detect such toxic and hazardous compounds in a more sensitive and safe way. Biosensor construction anticipates finding sensing elements able to detect TNT. As TNT can induce some physiological responses in E. coli, it may be useful to define the sensing elements from E. coli to detect TNT. An E. coli MG1655 genomic promoter library containing nearly 5,400 elements was constructed. Five elements, yadG, yqgC, aspC, recE, and topA, displayed high sensing specificity to TNT and its indicator compounds 1,3-DNB and 2,4-DNT. Based on this, a whole cell biosensor was constructed using E. coli, in which green fluorescent protein was positioned downstream of the five sensing elements via genetic fusion. The threshold value, detection time, EC200 value, and other aspects of five sensing elements were determined and the minimum responding concentration to TNT was 4.75 mg/L. According to the synthetic biology, the five sensing elements enriched the reservoir of TNT-sensing elements, and provided a more applicable toolkit to be applied in genetic routes and live systems of biosensors in future.
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Tan
authors.affiliation: College of Life Sciences, Anhui University, Hefei, China.
authors.forename: Naipeng
authors.initials: N
authors.lastname: Kan
authors.affiliation: College of Life Sciences, Jilin University, Changchun, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Jingyi
authors.initials: J
authors.lastname: Ling
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Guolong
authors.initials: G
authors.lastname: Qu
authors.affiliation: ShenYang Pharmaceutical University, Shenyang, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Jin
authors.affiliation: College of Life Sciences, Anhui University, Hefei, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China. liugang948784@163.com.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China. chenhp@nic.bmi.ac.cn.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D014303:Trinitrotoluene / Q000032:analysis* / Q000494:pharmacology
chemical_list: D029968:Escherichia coli Proteins; D014303:Trinitrotoluene; D049452:Green Fluorescent Proteins
keywords: Biosensor; GFP; Promoter; Synthetic biology; TNT