[1]王珏,徐佳迎,程粟裕,等.光谱法和分子对接研究3种兽医临床常用抗菌药对脲酶的影响及作用机制[J].南京农业大学学报,2020,43(4):690-697.[doi:10.7685/jnau.201909037]
 WANG Jue,XU Jiaying,CHENG Suyu,et al.The effects and mechanisms of three typical veterinary antimicrobials on the activity of urease studied by spectroscopic and molecular docking methods[J].Journal of Nanjing Agricultural University,2020,43(4):690-697.[doi:10.7685/jnau.201909037]
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光谱法和分子对接研究3种兽医临床常用抗菌药对脲酶的影响及作用机制()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
43卷
期数:
2020年4期
页码:
690-697
栏目:
生物与环境
出版日期:
2020-07-13

文章信息/Info

Title:
The effects and mechanisms of three typical veterinary antimicrobials on the activity of urease studied by spectroscopic and molecular docking methods
作者:
王珏 徐佳迎 程粟裕 赵鸽 蒋静艳
南京农业大学资源与环境科学学院, 江苏 南京 210095
Author(s):
WANG Jue XU Jiaying CHENG Suyu ZHAO Ge JIANG Jingyan
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China
关键词:
磺胺二甲嘧啶恩诺沙星四环素脲酶光谱法分子对接
Keywords:
sulfamethazine(SMZ)enrofloxacin(ENR)tetracycline(TC)ureasespectroscopic methodsmolecular docking
分类号:
X13
DOI:
10.7685/jnau.201909037
摘要:
[目的] 兽用抗菌药目前在畜牧业中广泛应用,大部分以其母体化合物形式随畜禽粪便排出体外,对水体和土壤环境造成严重污染,其对脲酶活性的影响与尿素氮利用率有关。本文旨在明确3种兽医临床常用抗菌药磺胺二甲嘧啶(SMZ)、恩诺沙星(ENR)和四环素(TC)对脲酶活性的影响及作用机制。[方法] 通过苯酚钠-次氯酸钠比色法,确定一定浓度范围内SMZ、ENR、TC(SMZ、TC:0~400 μmol·L-1;ENR:0~800 μmol·L-1)对脲酶活性的影响;通过荧光光谱结合紫外-可见吸收光谱法,确定SMZ、ENR、TC与脲酶相互作用的猝灭机制,并得到结合常数、结合位点数等参数信息;通过分子对接明确SMZ、ENR、TC在脲酶中的结合位点,并得到结合位点附近氨基酸残基信息和作用力信息。[结果] SMZ和ENR对脲酶活性有促进作用,而TC表现为低浓度促进、高浓度显著抑制的作用(P<0.05),影响程度随苯环数量增多而增强;SMZ、ENR、TC均能与脲酶形成1:1复合物,引起脲酶的内源荧光发生静态猝灭,并且改变了脲酶活性中心的氨基酸残基微环境,导致活性中心亲水性发生改变;SMZ、ENR、TC在脲酶活性中心与氨基酸残基结合产生氢键,除氢键作用力外,还有范德华力、去溶剂能和静电作用力发挥作用。[结论] SMZ、ENR和TC均与脲酶发生相互作用,可改变脲酶活性中心结构和氨基酸残基环境,但是TC对脲酶活性的影响与SMZ、ENR不同,表现为低浓度促进,高浓度抑制,而不同浓度的SMZ和ENR均可促进脲酶活性。
Abstract:
[Objectives] Veterinary antimicrobials are widely used in animal husbandry,most of which are discharged with livestock and poultry faeces in the form of their parent compounds,which cause serious pollution to water and soil environment. The effect of animal antimicrobials on urease activity was related to nitrogen use efficiency of urea. The purpose of this study was to investigate the effects of three typical veterinary antimicrobials,sulfamethazine(SMZ),enrofloxacin(ENR) and tetracycline(TC),on the activity of urease and its mechanism.[Methods] The effect of SMZ,ENR and TC on urease activity within a certain concentration range(SMZ,TC:0-400 μmol·L-1;ENR:0-800 μmol·L-1) was determined by sodium phenol sodium hypochlorite colorimetry. The quenching mechanism of the interaction of SMZ,ENR and TC with urease was determined by fluorescence spectroscopy combined with UV-Vis absorption spectroscopy,and the binding constant,binding site number and other parameter information were obtained. The binding sites of SMZ,ENR,TC in urease were identified by molecular docking,and the amino acid disability information and force information near the binding sites were obtained.[Results] SMZ and ENR could promote the activity of urease. Low concentration of TC could promote the activity of urease,and high concentration of TC significantly inhibited the activity of urease(P<0.05) and the influence degree increased with the increase of benzene ring number. SMZ,ENR,TC could form 1:1 complex with urease,resulting in static quenching of intrinsic fluorescence of urease and changing the micro-environment of amino acid residues in urease active center resulted in the hydrophilicity of the active site to change,thus affecting the activity of urease. SMZ,ENR,TC combined with amino acid residues in urease active center produced hydrogen bond. In addition to hydrogen bond,van der Waals force,desolvation energy,and electrostatic force also played a role.[Conclusions] SMZ,ENR and TC interacted with urease,which changed the structure of urease active center and amino acid residue environment,but the mechanism of interaction between TC and urease was different from that of SMZ and ENR,which led to SMZ and ENR promoting urease activity,while TC showed low concentration promotion and high concentration inhibition.

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备注/Memo

备注/Memo:
收稿日期:2019-09-23。
基金项目:国家自然科学基金项目(41675148,41375150)
作者简介:王珏,硕士研究生。
通信作者:蒋静艳,副教授,主要研究方向为环境污染与全球变化,E-mail:lilacjjy@njau.edu.cn。
更新日期/Last Update: 1900-01-01