| 信号分子N-酰基高丝氨酸内酯分析方法研究进展 |
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| 引用本文： | 生弘杰,宋 洋,卞永荣,柳广霞,刘总堂,蒋 新,王 芳.信号分子N-酰基高丝氨酸内酯分析方法研究进展[J].土壤学报,2016,53(4):832-844. |
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| 作者姓名： | 生弘杰 宋 洋 卞永荣 柳广霞 刘总堂 蒋 新 王 芳 |
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| 作者单位： | 1. 土壤与农业可持续发展国家重点实验室 中国科学院南京土壤研究所，南京 210008; 中国科学院大学，北京 100049;2. 土壤与农业可持续发展国家重点实验室 中国科学院南京土壤研究所,南京,210008 |
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| 基金项目： | 江苏省杰出青年基金（BK20150050）、国家重点基础研究发展计划（973计划）项目第五课题（2014CB441105）和国家自然科学基金项目 (21277148, 41301240, 41271327)资助 |
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| 摘 要： | 细菌能自发产生、释放特定的信号分子,并感知其浓度变化,调节微生物的群体行为,这一调控系统称为群体感应。革兰氏阴性菌的群体感应一般由N-酰基高丝氨酸内酯(Acylhomoserine lactones,AHLs)这类信号分子介导。在简要总结AHLs分子及其衍生物结构的基础上,结合近年来国内外研究进展,对AHLs的提取、鉴定和检测等分析测试方法进行了综述,旨在为建立土壤中AHLs的检测方法以及深入研究AHLs信号分子在土壤中的环境行为奠定基础。
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| 关 键 词： | N-酰基高丝氨酸内酯 色谱-质谱 生物传感器 核磁共振 酶联免疫 |
| 收稿时间： | 2015/10/17 0:00:00 |
| 修稿时间： | 2016/1/15 0:00:00 |
Advance in Study on Methods for Analysis of N-Acyl-Homoserine Lactones |
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| SHENG Hongjie,SONG Yang,BIAN Yongrong,LIU Guangxi,LIU Zongtang,JIANG Xin and WANG Fang.Advance in Study on Methods for Analysis of N-Acyl-Homoserine Lactones[J].Acta Pedologica Sinica,2016,53(4):832-844. |
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| Authors: | SHENG Hongjie SONG Yang BIAN Yongrong LIU Guangxi LIU Zongtang JIANG Xin and WANG Fang |
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| Institution: | State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences,State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences,State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences,State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences,State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences,State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences and State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences |
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| Abstract: | With the development of microbiology, Quorum sensing (QS), closely related to many physiological characteristics and activities, is getting more and more attention. QS in bacteria is a kind of mechanism for intracellular or intercellular communication in response to variation of the community in density, aiming at coordinating their population behavior and controlling gene expression. QS of Gram-negative bacteria is generally controlled by the Luxl/R-type information system, using N-acyl-homoserine lactones (AHLs) as signal molecules to regulate diverse physiological processes such as generation of exoenzyme and toxin by pathogenic bacteria, formation of biomembrane, bioluminescence, biosynthesis of antibiotic, bacteria motility and even degradation of organic pollutants.
How to rapidly and accurately detect and monitor variation of AHLs in environment and cells is an important basis for the study on quorum sensing. Therefore, an overall review is presented here on AHLs as representative signal molecules, summarizing their characteristics, structures, derivatives and different analyzing methods as well.
As the concentration of AHLs in the cell and the environment is usually very low, pre-concentration is necessary for its determination. For pre-treatment of liquid samples, Liquid-liquid extraction and solid-phase extraction are two common methods used to isolate AHLs from liquid matrix. The former is the most widely used one, while the latter has dual functions, concentration and purification, simultaneously. The identification of unknown signal molecules plays a vital role in understanding the information communication between microfloras. To identify structures of AHLs, the techniques of chromatograph-mass spectrometer, capillary zone electrophoresis/mass spectrometry (CZE/MS), Fourier transform ion cyclotron resonance mass spectrometry (FTICR/MS) and Nuclear magnetic resonance spectroscopy (NMR) are commonly used. To quantitatively analyze AHLs compounds, the techniques of biosensors, thin layer chromatography, high performance liquid chromatography (HPLC), radioactive isotope tracer and enzyme-linked immunosorbent assay (ELISA) are used. Each technique has its own advantages and disadvantages. Biosensors are very sensitive, however the analysis of each type of AHLs needs specific strain and plasmid. Thin layer chromatography, with a lower sensitivity, is only used to detect high concentration of AHLs. Radioactive isotope tracer is immune to interference of the external environment and hence makes the pretreatment of samples simple, however, this method can only be used to determine the total, rather than specific components of AHLs. HPLC is the most widely used to measure the content and structure of AHLs, but the samples for determination need to be pretreated for purification. Being low in cost, easy to operate and high in accuracy, ELISA has turned out to be a promising method, nevertheless, it could not be used to determine molecular structure of AHLs.
The determination and characterization of AHLs are very important to understand inter-microbial signal communication, however it is still a big challenge to determine AHLs qualitatively and quantitatively because they do not vary much in molecular structure and are often very low in concentration. Although the above mentioned analytic methods are quite high in sensitivity and selectivity, each has its own disadvantage. Therefore, it is advisable to combine some of these methods to bring their advantages into full play and complement each other in determining AHLs. At the same time, how to combine these methods into one that is capable of determining a variety of AHLs efficiently and rapidly shall be the focus of the researches in future.
On the other hand, the current methods are mainly focused on aqueous solution. In complex natural environments, such as soil and sediment, the existence of organic and inorganic compounds simultaneously presents a great challenge to extraction, purification and determination of N-acy-homoserine lactones. It is necessary to develop some efficient methods for qualitatively and quantitatively measuring AHLs in complex matrices. The determination will play an important role in unveiling the mechanism and action of microbial quorum sensing in the natural environment, especially the soil ecosystem. |
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| Keywords: | N-acyl-homoserine lactones Chromatography-mass spectrometry Biosensor Nuclear magnetic resonance Enzyme-linked immunosorbent |
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