微生物学报

Detection method	QS signals	LOD	Limitation	Application
Photoelectrochemical detection^[23]	AHLs	pmol	Poor universality; stability affected by environment	Real-time detection of specific signal molecules in fermentation workshop
β-D-glucuronide colorimetric assay^[37]	AIP-I	nmol	Easily interfered by other glycosides; narrow application range	Semi-quantification of AIP-I in soil and water environments
ELISA^[39]	AIP-IV	nmol	Antibody preparation required; susceptible to nonspecific adsorption from proteins	High-throughput semi-quantitative screening in strain fermentation broth
UPLC-MS^[38]	AHLs, AIPs, AI-2	fg	Weak matrix anti-interference	Semi-quantification of pure strain fermentation broth
HPLC-MS/MS^[43]	Slow analysis speed	Trace quantification in complex samples
LC-MS^[44]	Strict sample purity requirement	Rapid detection of multi-component
GC-MS^[44]	Short-chain-AHLs	ng	Only applicable to volatile and thermostable signal molecules	Qualitative analysis of short‑chain AHLs; headspace detection of fermentation broth
Reporter strain colorimetric assay^[40-41]	AHLs	μmol/L	Pretreatment needed for colored samples	QS agonist screening；qualitative detection of AHLs in strain fermentation broth
Fluorescent reporter strain^[45-46]	AHLs, AI-2	nmol/L	Long detection cycle; sample requires decolorization pretreatment	Mechanism visualization

Detection method	QS signals	LOD	Limitation	Application
Photoelectrochemical detection^[23]	AHLs	pmol	Poor universality; stability affected by environment	Real-time detection of specific signal molecules in fermentation workshop
β-D-glucuronide colorimetric assay^[37]	AIP-I	nmol	Easily interfered by other glycosides; narrow application range	Semi-quantification of AIP-I in soil and water environments
ELISA^[39]	AIP-IV	nmol	Antibody preparation required; susceptible to nonspecific adsorption from proteins	High-throughput semi-quantitative screening in strain fermentation broth
UPLC-MS^[38]	AHLs, AIPs, AI-2	fg	Weak matrix anti-interference	Semi-quantification of pure strain fermentation broth
HPLC-MS/MS^[43]	Slow analysis speed	Trace quantification in complex samples
LC-MS^[44]	Strict sample purity requirement	Rapid detection of multi-component
GC-MS^[44]	Short-chain-AHLs	ng	Only applicable to volatile and thermostable signal molecules	Qualitative analysis of short‑chain AHLs; headspace detection of fermentation broth
Reporter strain colorimetric assay^[40-41]	AHLs	μmol/L	Pretreatment needed for colored samples	QS agonist screening；qualitative detection of AHLs in strain fermentation broth
Fluorescent reporter strain^[45-46]	AHLs, AI-2	nmol/L	Long detection cycle; sample requires decolorization pretreatment	Mechanism visualization

QS system	Regulation methods	Applicable strains	Regulatory results	Validation methods
AIPs	Exogenous addition of AIP standards	Bifidobacterium, Bacillus subtilis, Lactobacillus sakei, L. plantarum, L. rhamnosus	Inhibition of biofilm formation in pathogenic bacteria^[50]	Crystal violet staining, scanning electron microscopy
Exogenous addition of cysteine, mg²⁺, and other cofactors	Promotion of intestinal colonization^[53-54]	16S rRNA gene sequencing, adhesion detection
Promotion of dietary fiber degradation^[53-54,72]	LC-MS, enzyme activity determination
AI-2	Exogenous addition of AI-2 precursors or standards	L. plantarum, Lactobacillus paralimentarius, B. amyloliquefaciens, L. rhamnosus, A. muciniphila	Promotion of self-biofilm formation^[49,70]	Crystal violet staining, scanning electron microscopy
Screening of mutant strains with enhanced AI-2 secretion ability through environmental stress	Promotion of mucus degradation^[7]	Glycosaminoglycan enzyme activity detection, adhesion detection
AHLs	Overexpression of synthetic genes LuxI or receptor LuxR	B. bifidum, Escherichia coli, L. crispatus, L. jensenii	Promotion of self-biofilm formation^[70]	Crystal violet staining, scanning electron microscopy
Exogenous addition of AHL or precursors like SAM	Regulation of carbohydrate metabolism and glycolysis^[73-74]	Enzyme activity determination, qPCR, isotope labeling and tracing
DSF	Exogenous addition of sucrose, branched-chain amino acids, and other DSF precursors	B. thetaiotaomicron	Promotion of dietary fiber degradation^[75]	Polysaccharide degrading enzyme activity determination, gel permeation chromatography, animal experiment

QS system	Regulation methods	Applicable strains	Regulatory results	Validation methods
AIPs	Exogenous addition of AIP standards	Bifidobacterium, Bacillus subtilis, Lactobacillus sakei, L. plantarum, L. rhamnosus	Inhibition of biofilm formation in pathogenic bacteria^[50]	Crystal violet staining, scanning electron microscopy
Exogenous addition of cysteine, mg²⁺, and other cofactors	Promotion of intestinal colonization^[53-54]	16S rRNA gene sequencing, adhesion detection
Promotion of dietary fiber degradation^[53-54,72]	LC-MS, enzyme activity determination
AI-2	Exogenous addition of AI-2 precursors or standards	L. plantarum, Lactobacillus paralimentarius, B. amyloliquefaciens, L. rhamnosus, A. muciniphila	Promotion of self-biofilm formation^[49,70]	Crystal violet staining, scanning electron microscopy
Screening of mutant strains with enhanced AI-2 secretion ability through environmental stress	Promotion of mucus degradation^[7]	Glycosaminoglycan enzyme activity detection, adhesion detection
AHLs	Overexpression of synthetic genes LuxI or receptor LuxR	B. bifidum, Escherichia coli, L. crispatus, L. jensenii	Promotion of self-biofilm formation^[70]	Crystal violet staining, scanning electron microscopy
Exogenous addition of AHL or precursors like SAM	Regulation of carbohydrate metabolism and glycolysis^[73-74]	Enzyme activity determination, qPCR, isotope labeling and tracing
DSF	Exogenous addition of sucrose, branched-chain amino acids, and other DSF precursors	B. thetaiotaomicron	Promotion of dietary fiber degradation^[75]	Polysaccharide degrading enzyme activity determination, gel permeation chromatography, animal experiment

益生菌群体感应系统的检测及功能调控

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王诣文 ¹ , 吴昊 ¹ , 陈丹蕾 ¹^,² , 财音青格乐 ¹^,² , 乔建军 ¹^,² , 李艳妮 ¹ , 吴胜波 ¹^,²

微生物学报 | 综述 2026,66(4): 1600-1615

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微生物学报 | 综述 2026, 66(4): 1600-1615

益生菌群体感应系统的检测及功能调控

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王诣文¹, 吴昊¹, 陈丹蕾¹^,², 财音青格乐¹^,², 乔建军¹^,², 李艳妮¹, 吴胜波¹^,²

作者信息

^1.天津大学合成生物与生物制造学院，合成生物技术全国重点实验室，系统生物工程教育部重点实验室，天津

^2.天津大学浙江研究院(绍兴)，浙江绍兴

Detection and functional regulation of the probiotic quorum sensing system

Yiwen WANG¹, Hao WU¹, Danlei CHEN¹^,², Qinggele CAIYIN¹^,², Jianjun QIAO¹^,², Yanni LI¹, Shengbo WU¹^,²

Affiliations

^1.State Key Laboratory of Synthetic Biology, Key Laboratory of Systems Bioengineering (Ministry of Education), School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin, China

^2.Zhejiang Institute of Tianjin University, Shaoxing, Zhejiang, China

出版时间: 2026-04-04 doi: 10.13343/j.cnki.wsxb.20250984

文章导航

摘要

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群体感应(quorum sensing, QS)是一种微生物通过分泌并感知信号分子进行通信，进而调控群体行为的机制，在益生菌生物被膜的形成与肠道定植过程中发挥重要作用。近年来，干预益生菌的QS已成为合成生物学领域的研究热点。本文综述了QS在益生菌中的分布情况，以及基于QS的益生功效干预策略；概述了目前已报道的益生菌QS系统，并对光电化学、色谱-质谱联用等检测益生菌QS的方法进行了总结与分析；重点阐述了干预益生菌QS的主要途径，包括QS激动剂的开发以及相关代谢通路的优化等。本文提出的基于QS的益生菌干预策略，为调控工程益生菌的功效提供了一种新型方案，对益生菌功能食品的开发与改进具有重要意义。

关键词

益生菌 / 群体感应 / 检测技术 / 干预策略 / 合成生物学

Abstract

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Quorum sensing (QS) is a communication mechanism through which microorganisms secrete and sense signal molecules to regulate mircobial population behaviors. QS plays important roles in biofilm formation and gut colonization of probiotics. In recent years, interfering with the QS of probiotics has become a trending research field of synthetic biology. In this review, we summarize the distribution of QS systems in probiotics and highlights interfering strategies designed to regulate probiotic functions. We summary currently identified QS systems in probiotics and their detection methods, such as photoelectrochemical assays and chromatography-mass spectrometry techniques. Meanwhile, this review outlines the QS interfering approaches for probiotics, including the use of QS agonists and the optimization of related metabolic pathways. Finally, the probiotic intervention strategy targeting QS is proposed in this paper, providing a novel approach for regulating the efficacy of engineered probiotics, which is of great significance for the development and improvement of probiotic functional foods.

Key words

probiotics / quorum sensing / detection technology / intervention strategy / synthetic biology

引用本文

王诣文, 吴昊, 陈丹蕾, 财音青格乐, 乔建军, 李艳妮, 吴胜波. 益生菌群体感应系统的检测及功能调控. 微生物学报, 2026 , 66 (4) : 1600 -1615 . DOI: 10.13343/j.cnki.wsxb.20250984

Yiwen WANG, Hao WU, Danlei CHEN, Qinggele CAIYIN, Jianjun QIAO, Yanni LI, Shengbo WU. Detection and functional regulation of the probiotic quorum sensing system[J]. Acta Microbiologica Sinica, 2026 , 66 (4) : 1600 -1615 . DOI: 10.13343/j.cnki.wsxb.20250984

正文

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细菌群体中生理变化的调节依赖于特定的细胞密度和生长阶段，这种细胞密度依赖性基因表达的现象称作群体感应(quorum sensing, QS)，最初因费氏另类弧菌(Aliivibrio fischeri)的生物发光现象而被发现^[1-4]。QS系统的提出使科学家们能够关注细菌的“社会”行为。然而，现有对QS的研究主要集中在病原菌上，对益生菌中QS的研究则知之甚少^[5-6]。益生菌可对宿主健康产生有益影响，其益生功效的发挥不仅依赖于摄入足够数量的活菌^[7]，更严重依赖于能否在宿主肠道等复杂环境中存活、定植并实现功能表达，这些过程均与QS系统密切相关^[8-12]。因此，解析益生菌的QS系统有助于理解其益生机制，为增强益生菌功效提供了全新的理论视角与技术路径。

益生菌的QS系统与其益生功效的实现密切相关，是当前微生态研究的前沿领域。现有研究表明，常见的益生菌株如乳酸杆菌、双歧杆菌等，均存在酰基高丝氨酸内酯(acyl-homoserine lactones, AHLs)、自诱导肽(autoinducing peptides, AI-2)、呋喃酰硼酸二酯(autoinducer-2, AIPs)、扩散性信号因子(diffusible signal factor, DSF)等不同类型的QS系统；其中，AHLs是革兰氏阴性菌种内特异性自诱导物，AIPs是革兰氏阳性菌肽类种内信号，AI-2是跨菌属的种间通用信号，DSF是革兰氏阴性菌的群体感应与致病性调控信号^[13-14]。益生菌的QS系统与益生功效直接相关，其中最典型的体现在生物被膜方面^[7,15]。益生菌通过形成生物被膜增强菌体对肠道上皮的黏附能力，这是在竞争激烈的肠道环境中占据优势生态位的重要方式。生物被膜构成了一个能够持续分泌抗菌物质、有机酸等代谢产物的微环境，有利于益生菌有效抑制病原菌、强化肠道屏障功能以及调节宿主免疫^[16-17]。此外，QS还参与调控胞外多糖的合成、应激耐受性以及生物活性物质的合成^[18-19]。除了对单菌的调控外，QS作为典型的微生物通讯机制，是理解多菌协同增效的有力证据。

对益生菌QS机制进行精准研究与有效干预，依赖于对信号分子等关键元件的高效检测^[20]。近年来，分析技术的发展使QS的原位实时定量检测成为可能。一方面，以色谱-质谱联用技术为代表的物理化学方法，如高效液相色谱-质谱、气相色谱-质谱等，凭借其高灵敏度、高分辨率和准确定量的优势，成为鉴定和定量发酵上清液、肠道内容物等复杂生物样本的标准技术^[21-22]；另一方面，基于QS调控原理构建的光电化学生物传感技术展现出巨大的应用潜力，这类传感器将生物识别元件与光电转换材料相结合，能够将QS的生物学信号转化为可测量的光或电信号，具有实时、快速甚至实现原位监测的优点^[23-24]。对这些先进检测方法的综合运用与对比分析，是精确绘制益生菌QS网络图谱、评估干预效果的关键技术基础。

本文系统梳理QS在益生菌领域的最新研究进展，详细总结QS在益生菌中的分布和检测情况，以及以激动剂和通路优化为核心的益生菌QS干预策略，深入探讨QS在开发高活性、高稳定性的功能性益生菌中的应用前景与技术挑战，为开发新一代微生态调控方案提供理论参考。

1 益生菌的QS系统

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QS系统在乳杆菌、双歧杆菌等革兰氏阳性菌，以及阿克曼氏菌、拟杆菌等革兰氏阴性菌中均有分布，但其信号分子类型与调控网络因菌种差异呈现显著多样性。现有研究表明，益生菌QS系统的功能多与代谢协同、宿主互作相关^[25]，与致病菌的毒力调控模式形成鲜明对比^[26-27]。

1.1 革兰氏阳性益生菌

革兰氏阳性菌如乳杆菌，信号分子以AIPs为主(图1A)，通过双组分系统(two-component system, TCS)传导信号。部分经典QS通路可通过调控鸟苷酸环化酶(diguanylate cyclase, DGC)、磷酸二酯酶(phosphodiesterase, PDE)活性，将胞间通讯信号转化为胞内环二鸟苷酸(cyclic diguanylate, c-di-GMP)的浓度变化，实现通讯信号到胞内表型的调控衔接。植物乳植杆菌(Lactiplantibacillus plantarum) WCFS1可通过CidA/CidB系统分泌第二信使c-di-GMP，进而调控自溶素基因表达，影响生物被膜形成与肠道定植能力^[28]。然而，植物乳植杆菌中此类胞内信号转导是否与QS系统相关联仍有待进一步验证。此外，AHLs类信号还能够通过激活宿主Toll样受体4 (Toll like receptors, TLR4)通路，抑制肥胖相关的慢性炎症^[29-30]。Turroni等^[31]发现两歧双歧杆菌(Bifidobacterium bifidum) PRL2010株也存在类似LuxI/LuxR的QS系统BifR/BifI，可以通过信号分子C8-HSL调控果糖-6-磷酸磷酸酮酶(fructose-6-phosphate phosphoketolase, F6PPK)基因表达，优化碳水化合物代谢路径(图1B)。此外，nis基因簇(nisABTCIPRKFEG)与NisRK共同构成了乳酸乳球菌的QS双组分系统，其调控的小五环肽Nisin兼具抑菌活性与AIP类QS信号分子功能。针对Nisin信号分子，本课题组Yuan等^[32]对其基于酶改造、底物修饰和序列挖掘的相关抗菌肽工程筛选和生产也做了梳理。

1.2 革兰氏阴性益生菌

革兰氏阴性菌如多形拟杆菌(Bacteroides thetaiotaomicron)可以利用DSF调控多糖利用基因puls的表达(图1C)。当菌群密度升高时，DSFs诱导BT_0415基因表达，促进木聚糖酶分泌，增强膳食纤维降解能力^[33]。此外，拟杆菌分泌的DSF信号分子可与肠道其他菌群形成交叉对话，通过AI-2/DSF混合信号实现菌群协作^[15,34]。阿克曼氏菌依赖AI-2信号分子和LuxS/LuxP通路调控Mucinase等黏液层降解酶的合成，转录组研究显示，高浓度的AI-2会使嗜黏蛋白阿克曼氏菌(Akkermansia muciniphila)黏蛋白降解基因簇AMUC_0005表达量上调2.3倍^[7]，这种由QS调控的黏液代谢可维持肠道屏障完整性(图1D)。

部分益生菌存在群体行为，但在基因组中并未检测出典型的QS基因簇。此类益生菌群体行为可能依赖乳酸等代谢产物的浓度效应而非特异性信号分子^[35]，抑或存在尚未被发现的QS系统，因此有待通过更实时精准的方法做进一步检测。此外，长双歧杆菌(Bifidobacterium longum)的类细菌素抑制物质合成系统(bacteriocin-like inhibitory substance, BLIS)是否与QS有关，以及是否直接参与宿主互作仍存在争议，需进一步验证^[36]。尽管目前益生菌中的QS系统已逐渐受到人们重视，但QS信号在复杂肠道菌群中的跨物种传导机制、益生菌QS系统与宿主TLR/NOD等免疫信号通路的交叉对话以及基于QS的益生菌工程化改造等领域仍有待后续进一步的研究。

2 益生菌QS的检测与鉴定

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益生菌QS干预的核心是通过调控信号分子合成或外源添加QS激动剂，增强益生菌自身QS介导的益生功能。对益生菌群体感应系统的挖掘，以及干预效果的评估依赖于精确的QS检测与验证方法。现有验证方法主要可分为信号分子检测、基因表达分析、表型验证3个方面。

2.1 信号分子检测

2.1.1 短肽类信号分子

益生菌以革兰氏阳性菌为主，这与其耐受胃肠道环境、易形成生物被膜等特性密切相关。目前对革兰氏阳性菌信号分子AIP进行检测的方法主要包括构建报告菌株以及酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)。Lubkowicz等^[37]对罗伊特氏黏液乳杆菌(Limosilactobacillus reuteri) DSM20016葡萄糖醛酸酶的表达通路进行编辑，通过AIP抑制gusA的转录，进而抑制葡萄糖醛酸酶产生。由于葡萄糖醛酸本身呈现为黄色，含有β-D-葡糖苷酸的溶液黄色变浅，通过比色的方法实现了μmol/L至nmol/L范围内AIP-1的定量分析。此外，对于在化学介质中不稳定的AIP类信号分子，也可以通过超高液相色谱质谱(ultra performance liquid chromatography-mass spectrometry, UPLC-MS)进行检测^[22,38]。Montagut等^[39]设计合成了对AIP-IV有高选择性的As380/AIPIVSBSA抗体，基于该抗体的ELISA检测灵敏度甚至高于质谱。

2.1.2 AHL类信号分子

随着阿克曼氏菌、拟杆菌等革兰氏阴性益生菌因其独特的代谢功能成为研究热点，益生菌的AHLs类信号分子的检测逐渐受到重视。利用生物传感器的显色反应对AHLs进行定性分析是一种快速高效的方法，如紫色色小杆菌(Chromobacterium violaceum) CV026与放射形农杆菌(Agrobacterium radiobacter)常用于初步检测AHLs^[40-41]。分子侧链的差异导致各种AHLs的极性不同，且内酯环m/z在质谱检测102处存在特征离子碎片，因此可使用高效液相色谱质谱(high-performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)对AHLs等信号分子进行精确定量。Morin等^[42]对利用HPLC-MS/MS检测AHLs类信号分子进行了改进，但依旧存在分析时间较长、操作繁琐等问题。

近年来，有机光电化学晶体管(organic photoelectrochemical transistor, OPECT)在生物传感领域展现出巨大的应用潜力。当信号分子与光电活性材料进行抗原-抗体结合等特异性相互作用，引起光电流、光电压变化就可以实现信号分子的快速定量检测。基于此，Ju等^[23]设计了一种基于金属有机骨架的CAU-17/Bi2S3 II型异质结的新型分离式OPECT适配传感器，实现了AHL类信号分子3-氧代-癸酰基高丝氨酸内酯(3-O-C10-HL)的快速检测，检测限达到了0.441 pmol/L。然而，PEC传感器的检测特异性依赖识别元件与目标信号分子的精准结合，目前识别元件的开发主要针对常见AHLs与AI-2类信号分子，对长链AHLs、革兰氏阳性益生菌分泌的AIPs覆盖度低，也难以实现多种信号分子的同时检测(表1)。

2.2 表型验证

研究表明，有些微生物中AHLs等信号分子的活性变化会影响特征代谢产物产量、菌体密度，以及光合作用、浮力、聚集状态等一系列关键理化与生理特性。AI-2在益生菌形成生物被膜中的作用被广泛报道，如解淀粉芽孢杆菌(Bacillus amyloliquefaciens) SQR9中可以通过AI-2促进生物被膜的形成，进而提高其在植物根部的定植能力^[47]。Liu等^[48]详细研究了LuxS/AI-2系统对类植物乳植杆菌(Lactobacillus paraplantarum) L-ZS9的生物被膜形成的促进作用机制，证明该菌的AI-2受到合成酶基因luxS的调控；过表达LuxS会增加AI-2的合成与分泌，促进L-ZS9生物被膜的形成，该效果受到SugE蛋白以及核酸内切酶的抑制，LuxS过表达还会影响AraC、PadR等与生物被膜形成有关的转录调节因子表达，进而影响与转运和膜功能相关的蛋白合成。

自然状态下的生物被膜通常由多种细菌共同组成，然而菌群中益生菌生物被膜的研究鲜有报道。近期研究人员发现益生菌可以抑制致病菌QS相关调控基因的表达，干扰致病菌生物被膜的形成，进而实现抗菌作用。例如，体外共培养的推测性模型显示，双歧杆菌益生菌分泌的AIPs可以与金黄色葡萄球菌(Staphylococcus aureus)的受体蛋白ArgC结合，从而下调生物被膜形成所必需的黏附素基因的表达^[49]。除了黏附相关基因，QS系统还可以调节curli、fim、fli等与益生菌运动相关的基因，通过改变益生菌运动性进而影响生物被膜的形成。Prouty等^[50]通过观察半固体琼脂培养基中细菌的扩散能力，发现QS调控细菌的群集运动。此外，Chatterjee等^[51]通过定量研究模式古菌的蛋白质组学，发现随着种群密度的增加，群体感应通路中与形状和运动相关的蛋白质丰度变化，从而使古菌从运动的杆状细胞过渡到非运动的盘状细胞。

James等^[52]发现植物乳植杆菌、瑞士乳杆菌以及唾液链球菌的组合对白色念珠菌生物膜形成具有抑制作用，但这种抑制与QS是否有关尚未得到解析。直到2024年，Zhang等^[53]发现枯草芽孢杆菌、鼠李糖乳酪杆菌(Lacticaseibacillus rhamnosus)和清酒乳杆菌等益生菌能分泌第二信号分子(c-di-GMP)，通过抑制与QS相关基因的表达实现对肺炎克雷伯氏菌定植与生物被膜形成的阻断。Zhang等^[54]发现某些食源性分子能与肠沙门氏菌(Salmonella enterica) LT2的受体LsrB结合，并改变生物被膜相关的生物表型，可能是潜在的QS干扰剂，例如粪臭素和2-氨基喹啉会抑制S. enterica LT2生物被膜的形成，而3-苯乳酸会促进LT2生物被膜的形成。食源或益生菌来源的代谢产物可以通过致病菌的QS系统促进致病菌的定植，也可通过精准干扰其QS通路抑制毒力因子表达，为靶向调控益生菌代谢通路、构建基于QS干预的益生策略提供了理论基础。

2.3 基因表达分析

将荧光蛋白等报告基因与群体感应调控启动子融合，可以通过检测报告基因的表达情况间接反映群体感应相关基因的表达水平，从而实现对群体感应的检测。Cao等^[55]构建了基于群体感应的自诱导动态调控系统，并利用荧光蛋白对启动子强度进行了表征。Taga等^[56]将AI-2的受体蛋白LuxP、青色荧光蛋白(cyan fluorescent protein, CFP)与黄色荧光蛋白(yellow fluorescent protein, YFP)组成CLPY融合蛋白；当AI-2与融合蛋白结合时，通过计算CFP与YFP之间荧光共振能量转移比例就可以实现对AI-2的定性分析。然而，这种方法难以进行定量分析，且需要提纯并标记受体蛋白与额外的荧光染料，成本较高。

群体感应的机制也可以通过信号通路互作进行验证。Maura等^[57]发现感应调控因子MvfR除了能够间接调控pqs和phnAB操作子的表达，还可以与基因组中包括群体感应调控因子lasR和rhlR在内的额外35个位点直接结合，并参与蛋白分泌、翻译和氧化应激反应。通过实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qPCR)检测hla、aprA等QS调控基因转录的mRNA表达水平，以及对转录组测序(RNA sequencing, RNA-seq)对益生菌全基因组表达谱进行分析，即可筛选出QS下游靶基因^[58]。Josephs-Spaulding等^[59]针对罗伊特氏黏液乳杆菌，系统整合了不同糖类、盐类、微生物共培养体系、母乳及果汁等50种独特环境条件的117组RNA-seq数据，成功识别出35个调控特定基因集的独立信号模块(imodulons)。与传统聚类分析相比，ICA技术能更精准捕捉基因簇内的细微调控关系，为通过转录组分析筛选益生菌中受群体感应调控的下游靶基因提供了新的技术思路。Zhang等^[60]利用染色质免疫共沉淀测序(chromatin immunoprecipitation sequencing, ChIP-seq)技术发现群体感应通过转录因子MupR可以调节微生物的碳代谢和氮代谢路径。qPCR具有靶向定量特性，可实现单个或多个QS基因表达模式的高效验证，适合实验室常规的基因表达差异分析。RNA-seq等高通量测序技术可以揭示QS信号分子诱导下的全基因组转录图谱，有利于挖掘未知的调控基因与代谢通路。值得注意的是，基因表达分析常与信号分子的定性定量检测及表型验证实验相结合，才能更全面地揭示QS的分子机制(图2)。

3 益生菌QS系统的干预

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基于对益生菌QS机制的深入理解与精准检测，靶向QS的干预策略应运而生，其核心目标是从正面强化益生菌的功能。与旨在抑制病原菌的“群体感应淬灭”策略相反，针对益生菌的干预更多采用激活QS通路的思路。例如，通过外源添加特定的信号分子类似物，或利用遗传工程手段优化其自身的QS代谢通路，提前启动或持续增强群体感应相关的有益群体行为。这些基于QS的精准干预策略超越了传统依靠补充活菌的低效模式，推动微生态调控领域向高效功能调控转变。

3.1 QS激动剂的开发

除了常见的群体感应信号分子外，一些天然提取物和人工合成的小分子化合物也具有群体感应激动剂的作用，进而影响细菌行为。本课题组Wu等^[61]针对影响QS活性的分子进行了系统分析，汇总了包含QS激动剂和拮抗剂在内的633种QS干扰剂，涵盖天然的和合成的QS类似物。

3.1.1 天然产物小分子

近期研究发现，许多天然产物在调节益生菌群体感应方面发挥着重要作用。植物根系分泌物的黄酮类化合物在植物与土壤益生菌的相互作用中意义重大，如Yu等^[62]针对植物根际促生益生菌荧光假单胞菌(Pseudomonas fluorescens) 2P24的研究发现，转录调控因子AefR能够感知芹菜素(apigenin)等黄酮类化合物，并通过抑制MexEF-OprN外排泵的表达来调控益生菌的QS系统；MexEF-OprN外排泵的作用是将AHLs群体感应信号分子排出细胞外，抑制该外排泵表达会使细胞内AHLs浓度增加，从而增强QS。然而，芹菜素等黄酮类化合物需借助肠道菌群的糖苷酶水解脱去糖基后才能被吸收利用，其生物转化效率受菌群结构影响较大。2024年，Zhu等^[63]在拟诺卡氏菌(Nocardiopsis sp.) ldbs0036中发现了一种新型信号分子α-吡喃酮(α-pyrone)，生物信息学分析发现，ldbs0036中吡喃酮合成基因上游存在一个多组分调控系统——conservon，敲除该系统的基因后吩嗪和吡喃酮的产量均受到影响，推测其用于接收和传递α-吡喃酮信号。拟诺卡氏菌能够合成抗生素等多种活性物质，是近几年广受关注的潜在益生菌。包含conservon系统的吡喃酮生物合成基因簇在拟诺卡氏菌属中广泛存在且高度保守，这意味着α-吡喃酮介导的QS调节机制在这类放线菌中普遍存在。

值得注意的是，α-吡喃酮作为拟诺卡氏菌的内源信号分子，仅在特定菌群微环境中的利用度较高，在复杂的肠道环境中易因稀释或降解而无法达到有效浓度。然而，从特异性来看，天然产物具有明确的靶向性，不会交叉激活铜绿假单胞菌等致病菌的群体感应通路，引发毒力因子表达等非预期调控效应。因此，挖掘新的天然产物激动剂具有重要的研究意义与产业化潜力，是未来QS调控的重要研究方向。此外，现有研究对激动剂在肠道中的生物利用度关注不足。大多数激动剂分子量大、亲水性强，难以穿透肠道黏液层，导致其在靶菌株附近的有效浓度较低。随着纳米载体包封以及靶向修饰技术的发展，未来或可以通过提高激动剂的肠道滞留时间和局部浓度来解决这一问题。

3.1.2 人工合成小分子

QS信号分子类似物起初主要是通过对天然信号分子结构进行修饰获得。例如，Geske等^[64]人工合成的N-(3-硝基-苯基乙酰酰基)-L-高丝氨酸内酯，与天然高丝氨酸内酯信号分子相比，该人工合成的QS类似物的侧链与受体的氢键相互作用更强。Bolitho等^[65]以CAI-1为基础进行结构修饰，得到了一系列具有激动活性的化合物，其中羟基酮官能团C3被NH₂取代的Am-CAI-1 (B11)是目前报道中活性最强的激动剂，EC₅₀达到了0.21 μmol/L。人工合成的信号分子类似物经结构修饰后与受体的结合活性显著提升，但长期暴露的生物安全性尚不明确。这类人工合成的分子在宿主肠道内的代谢途径、降解产物毒性，以及是否会引发免疫应答等问题仍缺乏体内验证(图3)。

3.2 QS相关生物表型的优化

在对益生菌的益生特性进行强化时通常包括肠道定植能力、代谢功能、免疫调节以及抑菌活性等。QS可以通过调控群体行为，实现益生菌在生物被膜形成、代谢协同2个方面的强化。

3.2.1 生物被膜形成能力的优化

现有研究表明，增强益生菌的肠道定植与黏附能力可以减少益生菌被宿主肠道蠕动排出的概率，从而增强益生菌对肠道菌群的调节作用以及对炎症性肠病的缓解作用^[66]。Holst等^[67]对比研究了植物乳植杆菌WCFS1和299v，发现植物乳植杆菌299v的黏附能力与编码甘露糖特异性黏附素msa基因的启动子区域有关。然而，目前针对益生菌的基因编辑技术缺乏普适性，尤其是像阿克曼氏菌这类新一代益生菌，其基因编辑方法尚未被充分开发。相比之下，益生菌的包埋技术具有通用性，Li等^[68]通过构建仙草多糖微凝胶递送体系，提升了益生菌的胃肠道耐受性和定植能力。然而，其弊端是包埋材料的生物相容性低，包埋效果不稳定，包埋菌活与代谢受限，肠道靶向释放效率不足。

益生菌通过寡肽、AHLs、AI-2信号分子进行种内与种间通讯，触发胞外多糖合成基因、黏附蛋白等生物被膜相关基因的表达，进而促进生物被膜的形成，并增强肠道定植稳定性。Deng等^[69]发现AI-2能够激活细胞外多糖(exopolysaccharide, EPS)合成基因和黏附相关蛋白MsA，进而显著增强鼠李糖乳酪杆菌生物被膜的结构稳定性。Sanchez等^[70]发现AHLs信号促进了益生菌细胞间聚集和胞外多糖分泌，形成致密的生物被膜结构，因此AHLs信号分子强度与卷曲乳杆菌(Lactobacillus crispatus)、詹氏乳杆菌(Lactobacillus jensenii)等阴道益生菌的生物被膜成熟度呈正相关。Ishida等^[71]通过核糖体工程改造鼠李糖乳酪杆菌，使其表面蛋白甘油醛-3-磷酸脱氢酶表达上调，并激活AI-2信号通路，进而促进生物被膜基质的释放。

3.2.2 代谢通路的优化

益生菌通过合成短链脂肪酸(short-chain fatty acids, SCFAs)、吲哚代谢物等有益产物，实现强化肠道屏障、参与代谢调控、介导肠-脑轴通讯等益生功能。许多研究从优化益生菌代谢层面实现了对其益生功能的调控。Qian等^[72]发现双歧杆菌可通过分泌吲哚-3-乳酸(indole-3-lactic acid, ILA)显著改善情绪。Kim等^[73]通过代谢物组学技术建立了代谢产物与tph1基因表达之间的互作量效关系，明确了L-乳酸、乙酸是短双歧杆菌调节肠道5-羟色胺(5-hydroxytryptamine, 5-HT)合成的关键效应物质；然而，由于代谢物与益生功能的因果关系难以确立、体外模拟环境的局限性、宿主体内微生物及相关代谢物繁多且互作复杂等问题，仅针对某一菌株的代谢物进行优化可能效果甚微。基于QS对代谢通路进行优化，在更好地模拟体内真实环境的同时，实现了益生菌代谢随种群密度变化的实时动态调控。

Gupta等^[74]对大肠杆菌AHL系统进行编辑，使菌体密度低时细胞优先表达糖酵解酶等生长相关的基因，AHL随菌体密度积累至临界浓度时与QS受体结合，激活下游启动子，定向合成葡萄糖酸。Cordeiro等^[75]对两歧双歧杆菌PRL2010中C8-HSL介导的BifR/BifI QS系统调控果糖-6-磷酸磷酸酮酶基因表达途径进行了综述，说明QS系统可以对碳水化合物代谢路径进行优化，提升对肠道复杂多糖的利用效率，间接增加SCFAs产量。此外，多形拟杆菌也可以通过DSF类信号分子调控多糖利用基因簇(puls)的表达，在菌群密度升高时协同分泌木聚糖酶，增强膳食纤维降解能力，为自身及宿主提供能量^[76] (表2)。

4 总结与展望

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益生菌群体感应系统在调控肠道菌群互作及其功能中发挥着关键作用^[77-78]，为长效益生菌制剂的研发提供了新思路。为更好地理解菌群QS的语言，本课题组Wu等^[79]在5株大肠杆菌中构建了QS语言“解释器”，模拟自然界微生物群落中的线性和环状QS通信网络，并开发了一个用于优化可调定向QSCN的动力学计算模型，填补了目前QSCN中传输多个QS信号的研究和合成空白。此外，Wu等^[80]汇总了“自下而上”组装功能微生物生态系统的各种串扰和正交调节模块，提出了一个包括功能规范、底盘选择、交互设计、系统构建、性能测试、建模分析和全局优化的全面设计-构建-测试-学习过程。实现了基于基因组规模模型的QS调控微生物生态系统的合理设计和优化^[81-82]。然而，目前针对益生菌群体感应系统如何精准调控其益生功效的分子机制与靶向干预策略，仍缺乏系统性的理论支撑与技术方案，有待进一步深入探索。

本文系统归纳了乳杆菌、双歧杆菌等革兰氏阳性益生菌，以及阿克曼氏菌、拟杆菌等革兰氏阴性菌中存在的群体感应现象与潜在的QS机制。在检测方法上，总结了现有基于生物传感、光电化学与色谱-质谱联用等技术在QS检测中的应用。研究表明，QS系统可通过促进生物被膜形成、优化代谢通路、增强环境应激耐受性等显著提升益生菌在肠道内的定植能力与益生功效。然而，当前益生菌的QS研究还存在一定挑战。

现有研究对各种益生菌中QS的分布挖掘不够完全，大部分的益生菌的QS机制解析不清晰，这是当前益生菌基于QS进行干预的主要困境^[83]。目前的研究多集中于乳酸杆菌、双歧杆菌等模式菌株，阿克曼氏菌、部分梭菌等许多具有潜在应用价值的益生菌是否拥有完整的QS系统，以及所采用的信号分子类型、遗传背景，均缺乏全面的基因组学与蛋白组学层面的研究证据。益生菌QS分布图谱的缺失使靶向QS的干预策略缺乏普适性的理论指导。对于存在QS现象的益生菌，具体的QS信号传导机制、调控网络及其与益生功能的因果关联的解析依旧十分困难。

当前各类QS检测技术虽已初步形成体系，但多以体外环境为主要应用场景，存在难以真实反映体内QS动态等局限。现有生物传感器多依赖于受体蛋白、适配体等生物识别元件与信号分子的特异性结合，易受温度等环境因素干扰^[84]。此外，细菌群体通讯中通常存在多种QS信号，多数传感器仅针对单一信号分子设计，导致菌群中复杂的QS网络难以解析^[85]。光电化学方法虽具有高灵敏度优势，但在检测电化学活性较弱的短链AHLs等分子时仍需依赖修饰电极以增强信号^[86]。金属纳米颗粒修饰电极易与样本中的还原性物质发生反应也会引入假阳性结果^[87]。色谱-质谱联用技术虽然具有良好的分离与鉴定能力，但检测样品需要进行复杂的前处理、检测周期较长，难以实现高通量分析与实时动态监测。更重要的是，上述方法均严重依赖对培养液等离体样本的处理与检测，无法复现宿主代谢物干扰、细胞间空间异质性等体内微环境对QS通路的调控作用，会导致体外结果与体内真实状态存在偏差^[88]。同时，QS信号分子在体外极易降解，如AHLs在酸碱环境中易水解或质子化^[89]，AIPs易被蛋白酶降解，AI-2对金属离子敏感^[90]等，这些都可能导致信号分子的检测值低于实际水平。因此，突破现有瓶颈的关键在于开发能够实现体内、实时、定量的检测新方法，为精准阐明菌群互作网络提供全新视角。

益生菌QS机制上的模糊性和检测验证的不准确性，直接导致干预的精准性与有效性不足。如果不能阐明益生菌QS代谢调控网络，通过代谢工程优化QS通路就会陷入盲目，甚至引发不可预见的代谢负担。因此，综合利用比较基因组学、转录组学、蛋白组学等多学科技术，系统地绘制QS在益生菌中遗传-调控-功能全景图谱是未来研究的首要任务。在此基础上，进一步结合益生元激动剂盲筛与QS网络靶向设计，(1) 通过高通量技术筛选如多酚类物质、特定结构寡糖等可能靶向益生菌QS系统的天然激动剂，系统解析其对QS信号分子合成与传递的调控规律；(2) 基于对QS通讯网络中关键节点的深入理解，开展激动剂的理性设计与优化，实现对益生菌定植能力、代谢活性及菌群互作效率的定向增强，是推动微生态调控由经验补充向机制驱动的关键。

基金

收起

浙江省“尖兵” “领雁”攻关计划(2025C01102)
国家自然科学基金(32300022)

参考文献

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排序方式：

[1]

Zhou

, Xu

, Zhang

, Liu

, Zhou

, Bi

, Han

, Tan

, Yuan

, Yang

. When the microbiome meets One Health principle: leading to the Holy Grail of biology and contributing to overall well-being and social sustainability[J]. iMetaOmics, 2024, 1(2): e30.

[2]

Lin

, Lee

, Yu

, Tsai

, She

, Li

, Liaw

. The QseEF two-component system-GlmY small RNA regulatory pathway controls swarming in uropathogenic Proteus mirabilis [J]. International Journal of Molecular Sciences, 2022, 23(1): 487.

[3]

Fang

, Wen

, Zhang

, Cao

. Dynamic regulation of efflux pump protein AcrA by quorum sensing system to improve free fatty acids production in Escherichia coli [J]. Biotechnology and Bioengineering, 2025, 122(9): 2499-2510.

[4]

Guo

, Wang

, Zheng

, Xue

, Wang

, Fang

, Zhang

. A WYL domain transcription factor regulates Lactiplantibacillus plantarum intestinal colonization via perceiving c-di-GMP[J]. Nature Communications, 2025, 16: 2193.

[5]

Sionov

, Steinberg

. Targeting the holy triangle of quorum sensing, biofilm formation, and antibiotic resistance in pathogenic bacteria[J]. Microorganisms, 2022, 10(6): 1239.

[6]

Lydick

, Mass

, Pepin

, Podicheti

, Klempic

, Rusch

, Ushijima

, Brown

, Salomon

, van Kessel

. Quorum sensing regulates virulence factors in the coral pathogen Vibrio coralliilyticus [J]. Applied and Environmental Microbiology, 2025, 91(2): e01143-24.

[7]

Yang

, Cong

. Gut microbiota-derived metabolites in the regulation of host immune responses and immune-related inflammatory diseases[J]. Cellular & Molecular Immunology, 2021, 18(4): 866-877.

[8]

Smith

WPJ

, Wucher

, Nadell

, Foster

. Bacterial defences: mechanisms, evolution and antimicrobial resistance[J]. Nature Reviews Microbiology, 2023, 21(8): 519-534.

[9]

Yang

, Yang

, Ye

, Zhang

, Nie

, Cheng

, Peng

, Zhang

, Liu

, Zhang

, Shi

. Physiologically inspired mucin coated Escherichia coli nissle 1917 enhances biotherapy by regulating the pathological microenvironment to improve intestinal colonization[J]. ACS Nano, 2022, 16(3): 4041-4058.

[10]

Pan

, Li

, Hou

, Lin

, Wang

, Pang

, Wang

, Liu

. Polymerization-mediated multifunctionalization of living cells for enhanced cell-based therapy[J]. Advanced Materials, 2021, 33(13): 2007379.

[11]

Jin

, Cao

, Gao

, Zhang

, Qian

, Zhang

, Mao

. Microenvironment-activated nanozyme-armed bacteriophages efficiently combat bacterial infection[J]. Advanced Materials, 2023, 35(30): 2301349.

[12]

Pan

, Jiang

, Wei

, Liu

, Zhang

, Gao

, Chen

, Yang

, Wang

, Yu

, Gan

. Ameba-inspired strategy enhances probiotic efficacy via prebound nutrient supply[J]. Nature Communications, 2025, 16: 1827.

[13]

Chai

, Ma

, Nong

, Mu

, Huang

. Dissecting Lu _x S/AI-2 quorum sensing system-mediated phenyllactic acid production mechanisms of Lactiplantibacillus plantarum L3[J]. Food Research International, 2023, 166: 112582.

[14]

Jacobi

, Grundler

, Hsieh

, Frick

, Adam

, Lamprecht

, Autenrieth

, Gregor

, Malfertheiner

. Quorum sensing in the probiotic bacterium Escherichia coli Nissle 1917 (Mutaflor)–evidence that furanosyl borate diester (AI-2) is influencing the cytokine expression in the DSS colitis mouse model[J]. Gut Pathogens, 2012, 4(1): 8.

[15]

, Yang

, Zhu

, Sun

, Wu

, Wang

, Lu

, He

, Tian

. Quorum-sensing signal DSF inhibits the proliferation of intestinal pathogenic bacteria and alleviates inflammatory response to suppress DSS-induced colitis in zebrafish[J]. Nutrients, 2024, 16(11): 1562.

[16]

Chen-Liaw

, Aggarwala

, Mogno

, Haifer

, Li

, Eggers

, Helmus

, Hart

, Wehkamp

, Lamousé- Smith

ESN

, Kerby

, Rey

, Colombel

, Kamm

, Olle

, Norman

, Menon

, Watson

, Crossette

, Terveer

, et al. Author correction: gut microbiota strain richness is species specific and affects engraftment[J]. Nature, 2025, 638(8050): E4.

[17]

Xiao

, Zou

, Li

, Song

, Lv

, Wang

, Tao

. Impact of quorum sensing signaling molecules in Gram-negative bacteria on host cells: current understanding and future perspectives[J]. Gut Microbes, 2022, 14: 2039048.

[18]

Seong

, Park

, Kim

, Yoo

, Kim

, Yoon

, Kim

, Lee

, Nam

, Hong

. Integrated multi-omics reveals different host crosstalk of atopic dermatitis-enriched Bifidobacterium longum strains[J]. npj Biofilms and Microbiomes, 2025, 11: 91.

[19]

Kareb

, Aïder

. Quorum sensing circuits in the communicating mechanisms of bacteria and its implication in the biosynthesis of bacteriocins by lactic acid bacteria: a review[J]. Probiotics and Antimicrobial Proteins, 2020, 12(1): 5-17.

[20]

Kumari

, Pasini

, Deo

, Flomenhoft

, Shashidhar

, Daunert

. Biosensing systems for the detection of bacterial quorum signaling molecules[J]. Analytical Chemistry, 2006, 78(22): 7603-7609.

[21]

, Li

, Tripathi

, Mitra

, Guerra

, Dash

, Weerasekera

, Makthal

, Shams

, Aggarwal

, Singh

, Gu

, Du

, Olsen

, LaRock

, Zhang

, Kumaraswami

. Engineered probiotic overcomes pathogen defences using signal interference and antibiotic production to treat infection in mice[J]. Nature Microbiology, 2024, 9(2): 502-513.

[22]

Verbeke

, Debunne

, Janssens

, Tack

, Wynendaele

, Rigole

, Coenye

, De Spiegeleer

. Detection and quantification of Enterococcus faecalis RNPP-type quorum sensing peptides in bacterial culture media by UHPLC-MS[J]. Journal of Pharmaceutical and Biomedical Analysis, 2018, 160: 55-63.

[23]

, Wang

, Wen

, Liu

, Wang

, Xiao

, Wang

, Ma

, Chi

. A dual-inhibition aptamer gated OPECT biosensor based on a MOF-derived CAU-17/Bi₂S₃ Z-scheme heterojunction for rapid detection of bacterial quorum sensing signal molecules[J]. Journal of Materials Chemistry C, 2025, 13(7): 3382-3391.

[24]

Chen

, Wang

, Zheng

, Hu

, Xia

. Emerging advances in biosensor technologies for quorum sensing signal molecules[J]. Analytical and Bioanalytical Chemistry, 2025, 417(1): 33-50.

[25]

, Luo

. Bacterial quorum-sensing systems and their role in intestinal bacteria-host crosstalk[J]. Frontiers in Microbiology, 2021, 12: 611413.

[26]

Medellin-Peña

, Wang

, Johnson

, Anand

, Griffiths

. Probiotics affect virulence-related gene expression in Escherichia coli O157:H7[J]. Applied and Environmental Microbiology, 2007, 73(13): 4259-4267.

[27]

Cella

, Coulson

, MacEachern

, Badr

, Ahmadi

, Tabatabaei

, Labbe

, Griffiths

. Probiotic disruption of quorum sensing reduces virulence and increases cefoxitin sensitivity in methicillin-resistant Staphylococcus aureus [J]. Scientific Reports, 2023, 13: 4373.

[28]

Fernández Ramírez

, Nierop Groot

, Smid

, Hols

, Kleerebezem

, Abee

. Role of cell surface composition and lysis in static biofilm formation by Lactobacillus plantarum WCFS1[J]. International Journal of Food Microbiology, 2018, 271: 15-23.

[29]

Coquant

, Aguanno

, Brot

, Belloir

, Delugeard

, Roger

, Pham

, Briand

, Moreau

, de Sordi

, Carrière

, Grill

, Thenet

, Seksik

. 3-oxo-C12: 2-HSL, quorum sensing molecule from human intestinal microbiota, inhibits pro-inflammatory pathways in immune cells via bitter taste receptors[J]. Scientific Reports, 2022, 12: 9440.

[30]

Jia

, Vianna

, Fukuda

, Berglund

, Liu

, Tao

, Sun

, Liu

, Harper

, Lee

, Scherer

, Elmquist

. Hepatocyte Toll-like receptor 4 regulates obesity-induced inflammation and insulin resistance[J]. Nature Communications, 2014, 5: 3878.

[31]

Turroni

, Duranti

, Milani

, Lugli

, van Sinderen

, Ventura

. Bifidobacterium bifidum: a key member of the early human gut microbiota[J]. Microorganisms, 2019, 7(11): 544.

[32]

Yuan

, Wu

, Tian

, Wang

, Wu

, Qiao

. Nisin-relevant antimicrobial peptides: synthesis strategies and applications[J]. Food & Function, 2024, 15(19): 9662-9677.

[33]

Sonnenburg

, Bäckhed

. Diet-microbiota interactions as moderators of human metabolism[J]. Nature, 2016, 535(7610): 56-64.

[34]

Sun

, He

, Brancaccio

, Yuan

, Riedel

. Bifidobacteria exhibit Lu _x S-dependent autoinducer 2 activity and biofilm formation[J]. PLoS One, 2014, 9(2): e88260.

[35]

Rul

, Ben-Yahia

, Chegdani

, Wrzosek

, Thomas

, Noordine

, Gitton

, Cherbuy

, Langella

, Thomas

. Impact of the metabolic activity of Streptococcus thermophilus on the colon epithelium of gnotobiotic rats[J]. Journal of Biological Chemistry, 2011, 286(12): 10288-10296.

[36]

Alessandri

, Ossiprandi

, MacSharry

, van Sinderen

, Ventura

. Bifidobacterial dialogue with its human host and consequent modulation of the immune system[J]. Frontiers in Immunology, 2019, 10: 2348.

[37]

Lubkowicz

, Ho

, Hwang

, Yew

, Lee

, Chang

. Reprogramming probiotic Lactobacillus reuteri as a biosensor for Staphylococcus aureus derived AIP-I detection[J]. ACS Synthetic Biology, 2018, 7(5): 1229-1237.

[38]

Junio

, Todd

, Ettefagh

, Ehrmann

, Kavanaugh

, Horswill

, Cech

. Quantitative analysis of autoinducing peptide I (AIP-I) from Staphylococcus aureus cultures using ultrahigh performance liquid chromatography–high resolving power mass spectrometry[J]. Journal of Chromatography B, 2013, 930: 7-12.

[39]

Montagut

, Acosta

, Albericio

, Royo

, Godoy-Tena

, Lacoma

, Prat

, Salvador

, Marco

. Direct quantitative immunochemical analysis of autoinducer peptide IV for diagnosing and stratifying Staphylococcus aureus infections[J]. ACS Infectious Diseases, 2022, 8(3): 645-656.

[40]

McClean

, Winson

, Fish

, Taylor

. Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones[J]. Microbiology, 1997, 143(12): 3703-3711.

[41]

Zhu

, Chai

, Zhong

, Li

. Agrobacterium bioassay strain for ultrasensitive detection of N-acylhomoserine lactone-type quorum-sensing molecules: detection of autoinducers in Mesorhizobium huakuii [J]. Applied and Environmental Microbiology, 2003, 69(11): 6949-6953.

[42]

Morin

, Grasland

, Vallée-Réhel

, Dufau

, Haras

. On-line high-performance liquid chromatography-mass spectrometric detection and quantification of N-acylhomoserine lactones, quorum sensing signal molecules, in the presence of biological matrices[J]. Journal of Chromatography A, 2003, 1002(1/2): 79-92.

[43]

Horáček

, Portillo

, Dhaubhadel

, Sung

, Readel

, Kučera

, Armstrong

. Comprehensive chiral GC-MS/MS and LC-MS/MS methods for identification and determination of N-acyl homoserine lactones[J]. Talanta, 2023, 253: 123957.

[44]

Andersen

, Heydorn

, Hentzer

, Eberl

, Geisenberger

, Christensen

, Molin

, Givskov

. gfp-based N-acyl homoserine-lactone sensor systems for detection of bacterial communication[J]. Applied and Environmental Microbiology, 2001, 67(2): 575-585.

[45]

Paul

, Kadosh

, Kushmaro

, Marks

. Microbead-encapsulated luminescent bioreporter screening of P. aeruginosa via its secreted quorum-sensing molecules[J]. Biosensors, 2024, 14(8): 383.

[46]

, Ni

, Li

, Du

, Sang

, Jiang

. Quorum sensing regulation in Microcystis aeruginosa: insights into AHL-mediated physiological processes and MC-LR production[J]. Science of the Total Environment, 2024, 919: 170867.

[47]

Xiong

, Liu

, Zhang

, Dong

, Zhang

, Liu

, Zhang

. Quorum sensing signal autoinducer-2 promotes root colonization of Bacillus velezensis SQR9 by affecting biofilm formation and motility[J]. Applied Microbiology and Biotechnology, 2020, 104(16): 7177-7185.

[48]

Liu

, Wu

, Zhang

, Li

. Overexpression of luxS promotes stress resistance and biofilm formation of Lactobacillus paraplantarum L-ZS9 by regulating the expression of multiple genes[J]. Frontiers in Microbiology, 2018, 9: 2628.

[49]

Kok

, Nyotohadi

. Biosurfactant potential and antiviral activity of multistrain probiotics[J]. Heliyon, 2024, 10(1): e22837.

[50]

Prouty

, Schwesinger

, Gunn

. Biofilm formation and interaction with the surfaces of gallstones by Salmonella spp.[J]. Infection and Immunity, 2002, 70(5): 2640-2649.

[51]

Chatterjee

, Consoli

, Schiller

, Winter

, McCallum

, Schulze

, Pohlschroder

. Quorum sensing mediates morphology and motility transitions in the model archaeon Haloferax volcanii [J]. mBio, 2025, 16(7): e00906-25.

[52]

James

, MacDonald

, Chanyi

, Cadieux

, Burton

. Inhibition of Candida albicans biofilm formation and modulation of gene expression by probiotic cells and supernatant[J]. Journal of Medical Microbiology, 2016, 65(4): 328-336.

[53]

Zhang

, Wang

, Dai

, Xiu

. The inhibition mechanism of co-cultured probiotics on biofilm formation of Klebsiella pneumoniae [J]. Journal of Applied Microbiology, 2024, 135(6): lxae138.

[54]

Zhang

, Chen

, Zhang

, Xiao

, Wu

, Caiyin

, Qiao

, Wu

. Mining and validating quorum sensing interference molecules from food-derived compounds for Salmonella Typhimurium [J]. Food & Function, 2025, 16(10): 3918-3933.

[55]

Cao

, Liu

, Zhang

, Mao

. P mutants with different promoting period and their application for quorum sensing regulated protein expression[J]. Food Science and Human Wellness, 2023, 12(5): 1841-1849.

[56]

Taga

, Xavier

. Methods for analysis of bacterial autoinducer-2 production[J]. Current Protocols in Microbiology, 2011, 23(1): 1.

[57]

Maura

, Hazan

, Kitao

, Ballok

, Rahme

. Evidence for direct control of virulence and defense gene circuits by the Pseudomonas aeruginosa quorum sensing regulator, MvfR[J]. Scientific Reports, 2016, 6: 34083.

[58]

Aguiar

VRC

, Castelli

, Single

, Bashirova

, Ramsuran

, Kulkarni

, Augusto

, Martin

, Gutierrez-Arcelus

, Carrington

, Meyer

. Comparison between qPCR and RNA-seq reveals challenges of quantifying HLA expression[J]. Immunogenetics, 2023, 75(3): 249-262.

[59]

Josephs-Spaulding

, Rajput

, Hefner

, Szubin

, Balasubramanian

, Li

, Zielinski

, Jahn

, Sommer

, Phaneuf

, Palsson

. Reconstructing the transcriptional regulatory network of probiotic L. reuteri is enabled by transcriptomics and machine learning[J]. mSystems, 2024, 9(3): e01257-23.

[60]

Zhang

, Li

, Nie

, Ma

, Tao

, Sun

, Tu

, Zhang

, Jia

, He

, Ge

. Quorum sensing coordinates carbon and nitrogen metabolism to optimize public goods production in Pseudomonas fluorescens 2P24[J]. Advanced Science, 2025, 12(12): 2412224.

[61]

, Liu

, Feng

, Yang

, Guo

, Qiao

. QSIdb: quorum sensing interference molecules[J]. Briefings in Bioinformatics, 2021, 22(4): bbaa218.

[62]

, Han

, Feng

, Hou

, Wang

, He

. The flavonoid-sensing regulator AefR is involved in modulating quorum sensing through repressing the MexEF-OprN efflux pump in Pseudomonas fluorescens [J]. mSystems, 2025, 10(3): e00915-24.

[63]

Zhu

, Cen

, Chen

, Shang

, Zhai

, Han

, Wang

, Chen

, Wei

, Han

. α-Pyrone mediates quorum sensing through the conservon system in Nocardiopsis sp.[J]. Microbiological Research, 2024, 285: 127767.

[64]

Geske

, O’Neill

, Blackwell

. N-phenylacetanoyl-L-homoserine lactones can strongly antagonize or superagonize quorum sensing in Vibrio fischeri [J]. ACS Chemical Biology, 2007, 2(5): 315-319.

[65]

Bolitho

, Perez

, Koch

, Ng

, Bassler

, Semmelhack

. Small molecule probes of the receptor binding site in the Vibrio cholerae CAI-1 quorum sensing circuit[J]. Bioorganic & Medicinal Chemistry, 2011, 19(22): 6906-6918.

[66]

Stephani

, Radulovic

, Niess

. Gut microbiota, probiotics and inflammatory bowel disease[J]. Archivum Immunologiae et Therapiae Experimentalis, 2011, 59(3): 161-177.

[67]

Holst

, Glenting

, Holmstrøm

, Israelsen

, Vrang

, Antonsson

, Ahrné

, Madsen

. Molecular switch controlling expression of the mannose-specific adhesin, msa, in Lactobacillus plantarum [J]. Applied and Environmental Microbiology, 2019, 85(10): e02954-18.

[68]

, Zhang

, Liu

, Xiao

, Stuart

MAC

, Hou

, Zhang

, Liang

, Li

, Wang

, Chen

, Li

. Natural phenolic-metal framework strengthened Mesona chinensis polysaccharides microgels for improved viability of probiotics to alleviate the liver injury and gut microbiota dysbiosis[J]. Advanced Functional Materials, 2024, 34(41): 2401064.

[69]

Deng

, Hou

, Valencak

, Luo

, Liu

, Wang

. AI-2/Lu _x S quorum sensing system promotes biofilm formation of Lactobacillus rhamnosus GG and enhances the resistance to enterotoxigenic Escherichia coli in germ-free zebrafish[J]. Microbiology Spectrum, 2022, 10(4): e00610-22.

[70]

Sanchez

, Alimiran

, Gandhi

, David

, Babayco

, Fiveash

, Garza

, Le

, Oldham

, Henderson

, Galloway

, Ventolini

. Comparison of biofilm growth and quorum sensing molecules in vaginal Lactobacillus species: a preliminary report[J]. Microbiology Research, 2024, 15(3): 1486-1493.

[71]

Ishida

, Fu

, Shigemori

, Kajikawa

, Tsukagoshi

, Sato

, Ogita

, Shimosato

. Ribosome-engineered Lacticaseibacillus rhamnosus strain GG exhibits cell surface glyceraldehyde-3-phosphate dehydrogenase accumulation and enhanced adhesion to human colonic mucin[J]. Applied and Environmental Microbiology, 2020, 86(20): e01448-20.

[72]

Qian

, Li

, Zhu

, Chen

, Lin

, Zhang

, Chen

, Wang

, Tian

. Bifidobacteria with indole-3-lactic acid-producing capacity exhibit psychobiotic potential via reducing neuroinflammation[J]. Cell Reports Medicine, 2024, 5(11): 101798.

[73]

Kim

, Jung

, Hwang

, Shin

. Gut microbiota indole-3-propionic acid mediates neuroprotective effect of probiotic consumption in healthy elderly: a randomized, double-blind, placebo-controlled, multicenter trial and in vitro study[J]. Clinical Nutrition, 2023, 42(6): 1025-1033.

[74]

Gupta

, Reizman

IMB

, Reisch

, Prather

KLJ

. Dynamic regulation of metabolic flux in engineered bacteria using a pathway-independent quorum-sensing circuit[J]. Nature Biotechnology, 2017, 35(3): 273-279.

[75]

Cordeiro

, RAS

Pirolla

, Persinoti

, Gozzo

, de Giuseppe

, Murakami

. N-glycan utilization by Bifidobacterium gut symbionts involves a specialist β-mannosidase[J]. Journal of Molecular Biology, 2019, 431(4): 732-747.

[76]

Devlin

, Marcobal

, Dodd

, Nayfach

, Plummer

, Meyer

, Pollard

, Sonnenburg

, Fischbach

. Modulation of a circulating uremic solute via rational genetic manipulation of the gut microbiota[J]. Cell Host & Microbe, 2016, 20(6): 709-715.

[77]

Liu

, Chen

, Lin

, Chen

, Hong

, Zhong

, Cai

, Wang

, Cui

. A multicellular self-organized probiotic platform for oral delivery enhances intestinal colonization[J]. Nature Communications, 2025, 16: 7060.

[78]

Guo

, Gao

, Wang

, Yuan

, Yin

, Wu

, Gao

, Zhu

, Zhang

, Wang

, Huang

, Kang

. An engineered probiotic consortium based on quorum-sensing for colorectal cancer immunotherapy[J]. Advanced Science, 2025, 12(46): e12744.

[79]

, Zhang

, Zhou

, Xu

, Yuan

, Wang

, Chen

, Liu

, Caiyin

, Wu

, Yang

, Zhao

, Qiao

. Design and analysis of quorum sensing language “Interpreter” ecosystem for microbial community[J]. Chemical Engineering Journal, 2024, 496: 153148.

[80]

, Zhou

, Dai

, Yang

, Qiao

. Assembly of functional microbial ecosystems: from molecular circuits to communities[J]. FEMS Microbiology Reviews, 2024, 48(6): fuae026.

[81]

Chen

, Qu

, Yang

, Li

, Yu

, Li

, Lu

, Yang

, Liu

, Wu

, Qiao

. Quorum sensing regulating the productivity and stability of cross-feeding cocultivation[J]. Chemical Engineering Journal, 2024, 502: 158167.

[82]

, Feng

, Liu

, Wu

, Qiu

, Ge

, Sun

, Hong

, Li

, Wang

, Yang

, Guo

, Qiao

. Machine learning aided construction of the quorum sensing communication network for human gut microbiota[J]. Nature Communications, 2022, 13: 3079.

[83]

Ayaz

, Gothalwal

. Effect of environmental factors on bacterial quorum sensing[J]. Cellular and Molecular Biology, 2014, 60(5): 46-50.

[84]

Zhang

, Lan

, Lu

. Overcoming major barriers to developing successful sensors for practical applications using functional nucleic acids[J]. Annual Review of Analytical Chemistry, 2022, 15: 151-171.

[85]

Vasudevan

, Srinivasan

, Neelakantan

, Rayappan

JBB

, Solomon

. Photoluminescence-based bioassay with cysteamine-capped TiO₂ nanoparticles for the selective recognition of N-acyl homoserine lactones[J]. Frontiers in Bioengineering and Biotechnology, 2021, 9: 750933.

[86]

Hermann

, Chen

, Wartchow

, Menke

, Gao

, Gleason

, Haynes

, Scott

, Petersen

, Gabriel

, Vu

, George

, Narayanan

, Li

, Qian

, Beatini

, Niu

, Gan

. Metal impurities cause false positives in high-throughput screening campaigns[J]. ACS Medicinal Chemistry Letters, 2013, 4(2): 197-200.

[87]

Dela Ahator

, Sagar

, Zhu

, Wang

, Zhang

. Nutrient availability and phage exposure alter the quorum-sensing and CRISPR-Cas-controlled population dynamics of Pseudomonas aeruginosa [J]. mSystems, 2022, 7(4): e00092-22.

[88]

, Chen

, Qiu

, Lin

, Tonokura

, Enami

. Decomposition mechanism of α-alkoxyalkyl-hydroperoxides in the liquid phase: temperature dependent kinetics and theoretical calculations[J]. Environmental Science: Atmospheres, 2022, 2(2): 241-251.

[89]

Churchill

MEA

, Chen

. Structural basis of acyl-homoserine lactone-dependent signaling[J]. Chemical Reviews, 2011, 111(1): 68-85.

[90]

Pereira

, Thompson

, Xavier

. AI-2-mediated signalling in bacteria[J]. FEMS Microbiology Reviews, 2013, 37(2): 156-181.

2026年第66卷第4期

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文章信息

doi: 10.13343/j.cnki.wsxb.20250984

接收时间：2025-12-28
首发时间：2026-04-14
出版时间：2026-04-04

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出版历史

收稿日期：2025-12-28
录用日期：2026-02-04

基金

“Pioneer” and “Leading Goose” Research and Development Program of Zhejiang Province(2025C01102)

浙江省“尖兵” “领雁”攻关计划(2025C01102)

National Natural Science Foundation of China(32300022)

国家自然科学基金(32300022)

作者信息

^1.天津大学合成生物与生物制造学院，合成生物技术全国重点实验室，系统生物工程教育部重点实验室，天津

^2.天津大学浙江研究院(绍兴)，浙江绍兴

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Detection method	QS signals	LOD	Limitation	Application
Photoelectrochemical detection^[23]	AHLs	pmol	Poor universality; stability affected by environment	Real-time detection of specific signal molecules in fermentation workshop
β-D-glucuronide colorimetric assay^[37]	AIP-I	nmol	Easily interfered by other glycosides; narrow application range	Semi-quantification of AIP-I in soil and water environments
ELISA^[39]	AIP-IV	nmol	Antibody preparation required; susceptible to nonspecific adsorption from proteins	High-throughput semi-quantitative screening in strain fermentation broth
UPLC-MS^[38]	AHLs, AIPs, AI-2	fg	Weak matrix anti-interference	Semi-quantification of pure strain fermentation broth
HPLC-MS/MS^[43]	Slow analysis speed	Trace quantification in complex samples
LC-MS^[44]	Strict sample purity requirement	Rapid detection of multi-component
GC-MS^[44]	Short-chain-AHLs	ng	Only applicable to volatile and thermostable signal molecules	Qualitative analysis of short‑chain AHLs; headspace detection of fermentation broth
Reporter strain colorimetric assay^[40-41]	AHLs	μmol/L	Pretreatment needed for colored samples	QS agonist screening；qualitative detection of AHLs in strain fermentation broth
Fluorescent reporter strain^[45-46]	AHLs, AI-2	nmol/L	Long detection cycle; sample requires decolorization pretreatment	Mechanism visualization

Detection method

QS signals

LOD

Limitation

Application

Photoelectrochemical detection^[23]

AHLs

pmol

Poor universality; stability affected by environment

Real-time detection of specific signal molecules in fermentation workshop

β-D-glucuronide colorimetric assay^[37]

AIP-I

nmol

Easily interfered by other glycosides; narrow application range

Semi-quantification of AIP-I in soil and water environments

ELISA^[39]

AIP-IV

nmol

Antibody preparation required; susceptible to nonspecific adsorption from proteins

High-throughput semi-quantitative screening in strain fermentation broth

UPLC-MS^[38]

AHLs, AIPs, AI-2

Weak matrix anti-interference

Semi-quantification of pure strain fermentation broth

HPLC-MS/MS^[43]

Slow analysis speed

Trace quantification in complex samples

LC-MS^[44]

Strict sample purity requirement

Rapid detection of multi-component

GC-MS^[44]

Short-chain-AHLs

Only applicable to volatile and thermostable signal molecules

Qualitative analysis of short‑chain AHLs; headspace detection of fermentation broth

Reporter strain colorimetric assay^[40-41]

AHLs

μmol/L

Pretreatment needed for colored samples

QS agonist screening；qualitative detection of AHLs in strain fermentation broth

Fluorescent reporter strain^[45-46]

AHLs, AI-2

nmol/L

Long detection cycle; sample requires decolorization pretreatment

Mechanism visualization

QS system	Regulation methods	Applicable strains	Regulatory results	Validation methods
AIPs	Exogenous addition of AIP standards	Bifidobacterium, Bacillus subtilis, Lactobacillus sakei, L. plantarum, L. rhamnosus	Inhibition of biofilm formation in pathogenic bacteria^[50]	Crystal violet staining, scanning electron microscopy
Exogenous addition of cysteine, mg²⁺, and other cofactors	Promotion of intestinal colonization^[53-54]	16S rRNA gene sequencing, adhesion detection
Promotion of dietary fiber degradation^[53-54,72]	LC-MS, enzyme activity determination
AI-2	Exogenous addition of AI-2 precursors or standards	L. plantarum, Lactobacillus paralimentarius, B. amyloliquefaciens, L. rhamnosus, A. muciniphila	Promotion of self-biofilm formation^[49,70]	Crystal violet staining, scanning electron microscopy
Screening of mutant strains with enhanced AI-2 secretion ability through environmental stress	Promotion of mucus degradation^[7]	Glycosaminoglycan enzyme activity detection, adhesion detection
AHLs	Overexpression of synthetic genes LuxI or receptor LuxR	B. bifidum, Escherichia coli, L. crispatus, L. jensenii	Promotion of self-biofilm formation^[70]	Crystal violet staining, scanning electron microscopy
Exogenous addition of AHL or precursors like SAM	Regulation of carbohydrate metabolism and glycolysis^[73-74]	Enzyme activity determination, qPCR, isotope labeling and tracing
DSF	Exogenous addition of sucrose, branched-chain amino acids, and other DSF precursors	B. thetaiotaomicron	Promotion of dietary fiber degradation^[75]	Polysaccharide degrading enzyme activity determination, gel permeation chromatography, animal experiment

QS system

Regulation methods

Applicable strains

Regulatory results

Validation methods

AIPs

Exogenous addition of AIP standards

Bifidobacterium, Bacillus subtilis, Lactobacillus sakei, L. plantarum, L. rhamnosus

Inhibition of biofilm formation in pathogenic bacteria^[50]

Crystal violet staining, scanning electron microscopy

Exogenous addition of cysteine, mg²⁺, and other cofactors

Promotion of intestinal colonization^[53-54]

16S rRNA gene sequencing, adhesion detection

Promotion of dietary fiber degradation^[53-54,72]

LC-MS, enzyme activity determination

AI-2

Exogenous addition of AI-2 precursors or standards

L. plantarum, Lactobacillus paralimentarius, B. amyloliquefaciens, L. rhamnosus, A. muciniphila

Promotion of self-biofilm formation^[49,70]

Crystal violet staining, scanning electron microscopy

Screening of mutant strains with enhanced AI-2 secretion ability through environmental stress

Promotion of mucus degradation^[7]

Glycosaminoglycan enzyme activity detection, adhesion detection

AHLs

Overexpression of synthetic genes LuxI or receptor LuxR

B. bifidum, Escherichia coli, L. crispatus, L. jensenii

Promotion of self-biofilm formation^[70]

Crystal violet staining, scanning electron microscopy

Exogenous addition of AHL or precursors like SAM

Regulation of carbohydrate metabolism and glycolysis^[73-74]

Enzyme activity determination, qPCR, isotope labeling and tracing

DSF

Exogenous addition of sucrose, branched-chain amino acids, and other DSF precursors

B. thetaiotaomicron

Promotion of dietary fiber degradation^[75]

Polysaccharide degrading enzyme activity determination, gel permeation chromatography, animal experiment