Bacterial cells rely on signaling molecules to communicate with others from the same species and induce certain genes in a process known as quorum sensing (QS). A common molecule is N-acyl homoserine lactone (AHL) whi...Bacterial cells rely on signaling molecules to communicate with others from the same species and induce certain genes in a process known as quorum sensing (QS). A common molecule is N-acyl homoserine lactone (AHL) which is responsible for the expression of virulence and other factors that allow the organisms to compete in a given environment. On the other hand, other bacteria produce certain enzymes such as AHL-lactonase that break down AHL molecules and prevent gene expression of these factors. The aim of this work was to examine the level of degradation of AHL molecules by AHL-lactonase in 62 Bacillus thuringiensis (Bt) strains isolated from Middle Tennessee, Mississippi, and Alabama. N-hexanoyl-homoserine lactone (C<sub>6</sub>-HSL) and N-3-oxo-hexanoyl homoserine lactone (3-oxo-C<sub>6</sub>-HSL), which cause Chromobacterium violaceum (CV026) to produce a purple pigment were tested at different concentrations to view the Bt lactonase activity. In addition, PCR was used to test for the presence of the lactonase gene. The results showed that among the 62 Bt strains, there were 58 that possessed the AHL-lactonase (aiiA) gene and 48 strains were able to degrade C<sub>6</sub>-HSL. At high concentrations of AHL, only 13 strains were able to completely degrade C6-HSL. In addition, degradation of 3-oxo-C<sub>6</sub>-HSL was weak compared to C<sub>6</sub>-HSL. The results also revealed that AHL lactonase was thermostable, and it was concluded that the level of degradation varies in Bt strains. Only 13 of the strains studied have potent inhibitory activity against C<sub>6</sub>-HSL, which may be good to be used in field applications to control agricultural pest.展开更多
The aim of this research was to detect the N-acyl homoserine lactones (AHLs) production and QseB/C gene of Aeromonas hydrophila. We analyzed the potentials of these isolates of Aeromonas hydrophila in causing biofilm ...The aim of this research was to detect the N-acyl homoserine lactones (AHLs) production and QseB/C gene of Aeromonas hydrophila. We analyzed the potentials of these isolates of Aeromonas hydrophila in causing biofilm formation, hemolysis, protease, and lipase. The antibiotic susceptibility of the 15 Aeromonas hydrophila isolates was also investigated. The detection of AHLs was carried out using the Chromobacterium violaceum strain CV026 as biosensors. The isolated strains were tested for the reaction of C. violaceum CV026 by cross-streaking on an agar plate. Production of AHLs was determined by the diffusing via the agar plates and the tinge of the biosensor strains. All isolated strains produced AHLs. A polymerase chain reaction (PCR) showed the isolated strains had qseB and qseC genes. Susceptibility tests of A. hydrophila isolates were administered against 25 different antibiotic disks representing 12 classes of antibiotics. The strains were highly resistant to β-Lactam with 96.7% showing resistibility, whereas 97.7% susceptibility was found towards Aminoglycoside class of the antibiotic used. 60% showed intermediate resistant to Polypeptide. 100% of the strains showed no resistant to Aminoglycoside, Polypeptide, Monobactam, and Carbapenems class of antibiotics. Each of the isolates was found to be associated with at least one virulent factor. Our results clearly demonstrated that there is a presence of QseB/C genes in A. hydrophila and also produces AHLs molecule and virulence factors. The investigated isolates showed the pathogenic potential of Aeromonas hydrophila which makes it a serious threat to public health.展开更多
The aim of this work was to inhibit biofilm formation by taking advantages of bacterial surface display technology in combination with cell membrane chromatography.A recombinant protein INPAidH was constructed by fusi...The aim of this work was to inhibit biofilm formation by taking advantages of bacterial surface display technology in combination with cell membrane chromatography.A recombinant protein INPAidH was constructed by fusing a quorum signal hydrolase AidH to the C-terminus of the ice nucleation protein(INP).Expression of INP-AidH was achieved on E.coli cell surface at an expression level of 30%of total membrane proteins.Activity of INP-AidH on cell membranes was confirmed in degrading the quorum signal C6-HSL as well as inhibiting bacterial biofilm.Immobilization of INP-AidH anchored cell membranes on silica gel particles was facilitated by taking advantages of cell membrane chromatography.The functionalized silica gel particles also exhibit activities in degrading C6-HSL and inhibiting bacterial biofilm.This article presents a new approach to prevent biofilm formation of silica-based materials.展开更多
This study presents a fast, accurate and sensitive technique using gas chromatography-mass spectrometry (GC-MS) for the identification and quantification of N-acyl homoserine lactones (AHLs) in the extracts of bacteri...This study presents a fast, accurate and sensitive technique using gas chromatography-mass spectrometry (GC-MS) for the identification and quantification of N-acyl homoserine lactones (AHLs) in the extracts of bacterial strain of Pseudomonas aeruginosa and sputum sample of a cystic fibrosis patient. This method involves direct separation and determination of AHLs by using GC-MS as simultaneous separation and characterization of AHLs were possible without any prior derivatiza-tion. Electron ionization resulted in a common fragmentation pattern with the most common fragment ion at m/z 143 and other minor peaks at 73, 57 and 43. The limit of detection for N-butanoyl, N-hexanoyl, N-octanoyl, N-decanoyl, N-dodecanoyl and N-tetradecanoyl homoserine lactones was 2.14, 3.59, 2.71, 2.10, 2.45 and 2.34 μg/L, respectively. The presence of AHLs in the culture of P. aeruginosa strain and spu-tum of a cystic fibrosis patient was achieved in selected ion monitoring (SIM) mode by using the prominent fragment at m/z 143.展开更多
The present study investigated the relationship between N-acyl-homoserine lactone(AHL)-based quorum sensing(QS) and the physico-chemical properties of aerobic granules.Stable mature granules were observed in SBR2 ...The present study investigated the relationship between N-acyl-homoserine lactone(AHL)-based quorum sensing(QS) and the physico-chemical properties of aerobic granules.Stable mature granules were observed in SBR2 and SBR3 with average diameters of 0.96,and1.49 mm,respectively. The sludge densities of aerobic granules in SBR2 and SBR3 were1.0246,and 1.0201 g/mL,respectively,which were higher than that of flocculent sludge in SBR1(1.0065 g/mL). The results showed that the activity of AHL-based QS in SBR2 and SBR3 amounted to 2.4- and 2.1-fold induction,however,that in SBR1 with flocculent sludge was1.6-fold induction. In addition,the results also showed that the activity of AHL-based QS in the three reactors rose in the feast condition,and then dropped with the consumption of substrate. However,the activity of AHL-based QS in these three reactors recovered again in prolonged starvation. Furthermore,the results showed that the enhancement of AHL-based QS favored the extracellular polymeric substance production of microorganisms in activated sludge. Thus,it could be concluded that aerobic granules showed higher AHL-based QS than flocculent sludge,which resulted from the higher sludge density of aerobic granules than flocculent sludge. AHL-based QS was related to the metabolism energy in the feast condition; however,in prolonged starvation,microorganisms would emit more AHL-like molecules to protect themselves to resist starvation. Moreover,the enhancement of AHL-based QS favored the EPS component productivity of the microorganisms in activated sludge,which contributed to maintain the aerobic granular structure.展开更多
N-acyl-homoserine lactones(AHLs) serve as quorum-sensing signals, which control a number of bacterial processes in many proteobacteria. Here we report the effects of exogenous short-chain AHL on the denitrifying pro...N-acyl-homoserine lactones(AHLs) serve as quorum-sensing signals, which control a number of bacterial processes in many proteobacteria. Here we report the effects of exogenous short-chain AHL on the denitrifying process of Paracoccus denitrificans, which are capable of aerobic and anaerobic growth by utilizing nitrate. The denitrification activity of these cells was monitored by measuring denitrification products(including nitrate, nitrite,and nitrous oxide), and the individual messenger ribonucleic acid(m RNA) levels of nitrate,nitrite, nitric oxide and nitrous oxide reductases. The results indicated that 2 μmol/L C6-homoserine lactone(HSL) has little effect on cell density under either anaerobic or aerobic culture conditions, and the nitrate reduction activity appeared slightly affected by N-hexanoyl-DL-homoserine lactone(C6-HSL). However, exogenous C6-HSL significantly affected the transcription of nitrite reductase and nitric oxide reductase genes in P.denitrificans regardless of the presence of oxygen, and N2O accumulation activity in P. denitrificans was suppressed by C6-HSL under aerobic condition. In contrast, exogenous C6-HSL stimulated the production of N2O under anaerobic condition, suggesting that the regulation of denitrification by quorum sensing may be important in N2O release.展开更多
Often as a result ofbiofilm formation, drinking water distribution systems (DWDS) are regularly faced with the problem of microbial contamination. Quorum sensing (QS) systems play a marked role in the regulation o...Often as a result ofbiofilm formation, drinking water distribution systems (DWDS) are regularly faced with the problem of microbial contamination. Quorum sensing (QS) systems play a marked role in the regulation of microbial biofilm formation; thus, inhibition of QS systems may provide a promising approach to biofilm formation control in DWDS. In the present study, 22 bacterial strains were isolated from drinking water-related environments. The following properties of the strains were investigated: bacterial biofilm formation capacity, QS signal molecule N-acyl-L-homoserine lactones (AHLs) production ability, and responses to AHLs and AHL analogs, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)- furanone (MX) and 2(5H)-furanone. Four AHLs were added to developed biofilms at dosages ranging from 0.1 nmol.L J to 100nmol.L1. As a result, the biofilm growth of more than 1/4 of the isolates, which included AHL producers and non-producers, were significantly promoted. Further, the biofilm biomasses were closely associated with respective AHLs concentrations. These results provided evidence to support the idea that AHLs play a definitive role in biofilm formation in many of the studied bacteria. Meanwhile, two AHLs analogs demon- strated unexpectedly minimal negative effects on biofilm formation. This suggested that, in order to find an applicable QS inhibition approach for biofilm control in DWDS, the testing and analysis of more analogs is needed.展开更多
Bacterial populations coexisting in the phyllosphere niche have important effects on plant health. Quorum sensing (QS) allows bacteria to communicate via diffusible signal molecules, but QS-dependent behaviors in ph...Bacterial populations coexisting in the phyllosphere niche have important effects on plant health. Quorum sensing (QS) allows bacteria to communicate via diffusible signal molecules, but QS-dependent behaviors in phyllosphere bacterial populations are poorly understood. We investigate the dense and diverse N-acyl-homoserine lactone (AHL)-producing phyllosphere bacteria living on tobacco leaf surfaces via a culture-dependent method and 16S rRNA gene sequencing. Our results indicated that approximately 7.9%-11.7% of the culturable leaf-associated bacteria have the ability to produce AHL based on the assays using whole-cell biosensors. Sequencing of the 16S rRNA gene assigned the AHL-producing strains to two phylogenetic groups, with Gammaproteobacteria (93%) as the predominant group, followed by Alphaproteobacteria. All of the AHL-producing Alphaproteobacteria were affiliated with the genus Rhizobium, whereas the AHL-producing bacteria belonging to the Gammaproteobacteria mainly fell within the genera Pseudomonas, Acinetobacter, Citrobacter, Enterobacter, Pantoea and Serratia. The bioassays of supernatant extracts revealed that a portion of the strains have a remarkable AHL profile for AHL induction activity using the two different biosensors, and one compound in the active extract of a representative isolate, NTL223, corresponded to 3-oxo-hexanoyl-homoserine lactone. A large population size and diversity of bacteria capable of AHL-driven QS were found to cohabit on leaves, implying that cross-communication based AHL-type QS may be common in the phyllosphere. Furthermore, this study provides a general snapshot of a potential valuable application of AHL-producing bacteria inhabiting leaves for their presumable ecological roles in the phyllosphere.展开更多
文摘Bacterial cells rely on signaling molecules to communicate with others from the same species and induce certain genes in a process known as quorum sensing (QS). A common molecule is N-acyl homoserine lactone (AHL) which is responsible for the expression of virulence and other factors that allow the organisms to compete in a given environment. On the other hand, other bacteria produce certain enzymes such as AHL-lactonase that break down AHL molecules and prevent gene expression of these factors. The aim of this work was to examine the level of degradation of AHL molecules by AHL-lactonase in 62 Bacillus thuringiensis (Bt) strains isolated from Middle Tennessee, Mississippi, and Alabama. N-hexanoyl-homoserine lactone (C<sub>6</sub>-HSL) and N-3-oxo-hexanoyl homoserine lactone (3-oxo-C<sub>6</sub>-HSL), which cause Chromobacterium violaceum (CV026) to produce a purple pigment were tested at different concentrations to view the Bt lactonase activity. In addition, PCR was used to test for the presence of the lactonase gene. The results showed that among the 62 Bt strains, there were 58 that possessed the AHL-lactonase (aiiA) gene and 48 strains were able to degrade C<sub>6</sub>-HSL. At high concentrations of AHL, only 13 strains were able to completely degrade C6-HSL. In addition, degradation of 3-oxo-C<sub>6</sub>-HSL was weak compared to C<sub>6</sub>-HSL. The results also revealed that AHL lactonase was thermostable, and it was concluded that the level of degradation varies in Bt strains. Only 13 of the strains studied have potent inhibitory activity against C<sub>6</sub>-HSL, which may be good to be used in field applications to control agricultural pest.
文摘The aim of this research was to detect the N-acyl homoserine lactones (AHLs) production and QseB/C gene of Aeromonas hydrophila. We analyzed the potentials of these isolates of Aeromonas hydrophila in causing biofilm formation, hemolysis, protease, and lipase. The antibiotic susceptibility of the 15 Aeromonas hydrophila isolates was also investigated. The detection of AHLs was carried out using the Chromobacterium violaceum strain CV026 as biosensors. The isolated strains were tested for the reaction of C. violaceum CV026 by cross-streaking on an agar plate. Production of AHLs was determined by the diffusing via the agar plates and the tinge of the biosensor strains. All isolated strains produced AHLs. A polymerase chain reaction (PCR) showed the isolated strains had qseB and qseC genes. Susceptibility tests of A. hydrophila isolates were administered against 25 different antibiotic disks representing 12 classes of antibiotics. The strains were highly resistant to β-Lactam with 96.7% showing resistibility, whereas 97.7% susceptibility was found towards Aminoglycoside class of the antibiotic used. 60% showed intermediate resistant to Polypeptide. 100% of the strains showed no resistant to Aminoglycoside, Polypeptide, Monobactam, and Carbapenems class of antibiotics. Each of the isolates was found to be associated with at least one virulent factor. Our results clearly demonstrated that there is a presence of QseB/C genes in A. hydrophila and also produces AHLs molecule and virulence factors. The investigated isolates showed the pathogenic potential of Aeromonas hydrophila which makes it a serious threat to public health.
基金Funded by the National Natural Science Foundation of China(No.31771032)。
文摘The aim of this work was to inhibit biofilm formation by taking advantages of bacterial surface display technology in combination with cell membrane chromatography.A recombinant protein INPAidH was constructed by fusing a quorum signal hydrolase AidH to the C-terminus of the ice nucleation protein(INP).Expression of INP-AidH was achieved on E.coli cell surface at an expression level of 30%of total membrane proteins.Activity of INP-AidH on cell membranes was confirmed in degrading the quorum signal C6-HSL as well as inhibiting bacterial biofilm.Immobilization of INP-AidH anchored cell membranes on silica gel particles was facilitated by taking advantages of cell membrane chromatography.The functionalized silica gel particles also exhibit activities in degrading C6-HSL and inhibiting bacterial biofilm.This article presents a new approach to prevent biofilm formation of silica-based materials.
文摘This study presents a fast, accurate and sensitive technique using gas chromatography-mass spectrometry (GC-MS) for the identification and quantification of N-acyl homoserine lactones (AHLs) in the extracts of bacterial strain of Pseudomonas aeruginosa and sputum sample of a cystic fibrosis patient. This method involves direct separation and determination of AHLs by using GC-MS as simultaneous separation and characterization of AHLs were possible without any prior derivatiza-tion. Electron ionization resulted in a common fragmentation pattern with the most common fragment ion at m/z 143 and other minor peaks at 73, 57 and 43. The limit of detection for N-butanoyl, N-hexanoyl, N-octanoyl, N-decanoyl, N-dodecanoyl and N-tetradecanoyl homoserine lactones was 2.14, 3.59, 2.71, 2.10, 2.45 and 2.34 μg/L, respectively. The presence of AHLs in the culture of P. aeruginosa strain and spu-tum of a cystic fibrosis patient was achieved in selected ion monitoring (SIM) mode by using the prominent fragment at m/z 143.
基金supported by the National Natural Science Found of China (No.51078036)
文摘The present study investigated the relationship between N-acyl-homoserine lactone(AHL)-based quorum sensing(QS) and the physico-chemical properties of aerobic granules.Stable mature granules were observed in SBR2 and SBR3 with average diameters of 0.96,and1.49 mm,respectively. The sludge densities of aerobic granules in SBR2 and SBR3 were1.0246,and 1.0201 g/mL,respectively,which were higher than that of flocculent sludge in SBR1(1.0065 g/mL). The results showed that the activity of AHL-based QS in SBR2 and SBR3 amounted to 2.4- and 2.1-fold induction,however,that in SBR1 with flocculent sludge was1.6-fold induction. In addition,the results also showed that the activity of AHL-based QS in the three reactors rose in the feast condition,and then dropped with the consumption of substrate. However,the activity of AHL-based QS in these three reactors recovered again in prolonged starvation. Furthermore,the results showed that the enhancement of AHL-based QS favored the extracellular polymeric substance production of microorganisms in activated sludge. Thus,it could be concluded that aerobic granules showed higher AHL-based QS than flocculent sludge,which resulted from the higher sludge density of aerobic granules than flocculent sludge. AHL-based QS was related to the metabolism energy in the feast condition; however,in prolonged starvation,microorganisms would emit more AHL-like molecules to protect themselves to resist starvation. Moreover,the enhancement of AHL-based QS favored the EPS component productivity of the microorganisms in activated sludge,which contributed to maintain the aerobic granular structure.
基金supported by the National Natural Science Foundation of China(Nos.41371266,41501250,21377157)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB15030101)
文摘N-acyl-homoserine lactones(AHLs) serve as quorum-sensing signals, which control a number of bacterial processes in many proteobacteria. Here we report the effects of exogenous short-chain AHL on the denitrifying process of Paracoccus denitrificans, which are capable of aerobic and anaerobic growth by utilizing nitrate. The denitrification activity of these cells was monitored by measuring denitrification products(including nitrate, nitrite,and nitrous oxide), and the individual messenger ribonucleic acid(m RNA) levels of nitrate,nitrite, nitric oxide and nitrous oxide reductases. The results indicated that 2 μmol/L C6-homoserine lactone(HSL) has little effect on cell density under either anaerobic or aerobic culture conditions, and the nitrate reduction activity appeared slightly affected by N-hexanoyl-DL-homoserine lactone(C6-HSL). However, exogenous C6-HSL significantly affected the transcription of nitrite reductase and nitric oxide reductase genes in P.denitrificans regardless of the presence of oxygen, and N2O accumulation activity in P. denitrificans was suppressed by C6-HSL under aerobic condition. In contrast, exogenous C6-HSL stimulated the production of N2O under anaerobic condition, suggesting that the regulation of denitrification by quorum sensing may be important in N2O release.
文摘Often as a result ofbiofilm formation, drinking water distribution systems (DWDS) are regularly faced with the problem of microbial contamination. Quorum sensing (QS) systems play a marked role in the regulation of microbial biofilm formation; thus, inhibition of QS systems may provide a promising approach to biofilm formation control in DWDS. In the present study, 22 bacterial strains were isolated from drinking water-related environments. The following properties of the strains were investigated: bacterial biofilm formation capacity, QS signal molecule N-acyl-L-homoserine lactones (AHLs) production ability, and responses to AHLs and AHL analogs, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)- furanone (MX) and 2(5H)-furanone. Four AHLs were added to developed biofilms at dosages ranging from 0.1 nmol.L J to 100nmol.L1. As a result, the biofilm growth of more than 1/4 of the isolates, which included AHL producers and non-producers, were significantly promoted. Further, the biofilm biomasses were closely associated with respective AHLs concentrations. These results provided evidence to support the idea that AHLs play a definitive role in biofilm formation in many of the studied bacteria. Meanwhile, two AHLs analogs demon- strated unexpectedly minimal negative effects on biofilm formation. This suggested that, in order to find an applicable QS inhibition approach for biofilm control in DWDS, the testing and analysis of more analogs is needed.
基金supported by the National Natural Science Foundation of China (No.21177145, 20977106)the Key Laboratory of Pollution Processes and Environmental Criteria (Nankai University), Ministry of Education (No.KL-PPEC-2010-3)
文摘Bacterial populations coexisting in the phyllosphere niche have important effects on plant health. Quorum sensing (QS) allows bacteria to communicate via diffusible signal molecules, but QS-dependent behaviors in phyllosphere bacterial populations are poorly understood. We investigate the dense and diverse N-acyl-homoserine lactone (AHL)-producing phyllosphere bacteria living on tobacco leaf surfaces via a culture-dependent method and 16S rRNA gene sequencing. Our results indicated that approximately 7.9%-11.7% of the culturable leaf-associated bacteria have the ability to produce AHL based on the assays using whole-cell biosensors. Sequencing of the 16S rRNA gene assigned the AHL-producing strains to two phylogenetic groups, with Gammaproteobacteria (93%) as the predominant group, followed by Alphaproteobacteria. All of the AHL-producing Alphaproteobacteria were affiliated with the genus Rhizobium, whereas the AHL-producing bacteria belonging to the Gammaproteobacteria mainly fell within the genera Pseudomonas, Acinetobacter, Citrobacter, Enterobacter, Pantoea and Serratia. The bioassays of supernatant extracts revealed that a portion of the strains have a remarkable AHL profile for AHL induction activity using the two different biosensors, and one compound in the active extract of a representative isolate, NTL223, corresponded to 3-oxo-hexanoyl-homoserine lactone. A large population size and diversity of bacteria capable of AHL-driven QS were found to cohabit on leaves, implying that cross-communication based AHL-type QS may be common in the phyllosphere. Furthermore, this study provides a general snapshot of a potential valuable application of AHL-producing bacteria inhabiting leaves for their presumable ecological roles in the phyllosphere.