当前位置: X-MOL 学术ACS Sens. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
2D Strategy for the Construction of an Enzyme-Activated NIR Fluorophore Suitable for the Visual Sensing and Profiling of Homologous Nitroreductases from Various Bacterial Species
ACS Sensors ( IF 8.2 ) Pub Date : 2021-09-01 , DOI: 10.1021/acssensors.1c01216
Tao Liu 1, 2 , Yifei Wang 1 , Lei Feng 1 , Xiangge Tian 1 , Jingnan Cui 2 , Zhenlong Yu 1 , Chao Wang 1 , Baojing Zhang 1 , Tony D James 3, 4 , Xiaochi Ma 1, 5
Affiliation  

Nitroreductases (NTRs) mediate the reduction of nitroaromatic compounds to the corresponding nitrite, hydroxylamine, or amino derivatives. The activity of NTRs in bacteria facilitates the metabolic activation and antibacterial activity of 5-nitroimidazoles. Therefore, NTR activity correlates with the drug susceptibility and resistance of pathogenic bacteria. As such, it is important to develop a rapid and visual assay for the real-time sensing of bacterial NTRs for the evaluation and development of antibiotics. Herein, an activatable near-infrared fluorescent probe (HC–NO2) derived from a hemicyanine fluorophore was designed and developed based on two evaluation factors, including the calculated partition coefficient (Clog P) and fluorescence wavelength. Using HC–NO2 as the special substrate of NTRs, NTR activity can be assayed efficiently, and then, bacteria can be imaged based on the detection of NTRs. More importantly, a sensitive in-gel assay using HC–NO2 has been developed to selectively identify NTRs and sensitively determine NTR activity. Using the in-gel assay, NTRs from various bacterial species have been profiled visually from the “fluorescence fingerprints”, which facilitates the rapid identification of NTRs from bacterial lysates. Thus, various homologous NTRs were identified from three metronidazole-susceptible bacterial species as well as seven unsusceptible species, which were confirmed by the whole-genome sequence. As such, the evaluation of NTRs from different bacterial species should help improve the rational usage of 5-nitroimidazole drugs as antibiotics.

中文翻译:


构建适用于各种细菌同源硝基还原酶视觉传感和分析的酶激活近红外荧光团的二维策略



硝基还原酶 (NTR) 介导硝基芳香族化合物还原为相应的亚硝酸盐、羟胺或氨基衍生物。细菌中NTR的活性促进5-硝基咪唑的代谢活化和抗菌活性。因此,NTR活性与病原菌的药物敏感性和耐药性相关。因此,开发一种快速、可视化的实时检测细菌 NTR 的方法对于抗生素的评估和开发非常重要。在此,基于两个评估因素,包括计算的分配系数(Clog P )和荧光波长,设计和开发了一种源自半花青荧光团的可激活近红外荧光探针( HC–NO 2 )。利用HC–NO 2作为NTRs的特殊底物,可以有效地测定NTRs活性,然后根据NTRs的检测对细菌进行成像。更重要的是,我们开发了一种使用HC-NO 2的灵敏凝胶内测定法,可以选择性地识别 NTR 并灵敏地测定 NTR 活性。使用凝胶内测定,可以从“荧光指纹”中直观地描绘出来自不同细菌物种的 NTR,这有助于从细菌裂解物中快速识别 NTR。因此,从三种甲硝唑敏感细菌物种以及七种不敏感细菌物种中鉴定出各种同源NTR,并通过全基因组序列证实。因此,对不同菌种的NTR进行评价有助于提高5-硝基咪唑类药物作为抗生素的合理使用。
更新日期:2021-09-24
down
wechat
bug