Understanding aquatic pathogen in sediments or aquacultural water is crucial to protect public health from soilborne and waterborne diseases.Quorum sensing(QS)was increasingly reported in biological wastewater treatme...Understanding aquatic pathogen in sediments or aquacultural water is crucial to protect public health from soilborne and waterborne diseases.Quorum sensing(QS)was increasingly reported in biological wastewater treatment processes because of their inherent roles in biofilm development,bacterial aggregation and so on.The widely QS signals was Antoinducer-2(AI-2),primarily involved to allow the possibility of interspecies communication.However,the cellular components that mediate the response of Streptococcus agalactiae to AI-2 have not been fully characterized.Analysis of the complete genome sequence of S.agalactiae indi-cated that its RbsB protein has similarity to Escherichia coli LsrB and Aggregatibacter actinomycetemcomitans RbsB proteins that bind AI-2.We hypothesized that RbsB protein mediates quorum sensing signal uptake via interaction with AI-2.To evaluate the regulatory effect of RbsB on QS system,the recombinant plasmid pGEX-6p-1-RbsB was constructed and RbsB protein was purified with GST-tag.To further elucidate the role of RbsB protein binding to DPD(AI-2 precursor dihydroxypentanedione),the systemati-cally throughput circular dichroism(CD)spectroscopy,isothermal titration calorimetry200(ITC200)and molecular docking methods were employed.The high expression of soluble RbsB protein with molecular weight of 33 kDa was obtained.The thermodynamics results(ΔH<0,ΔS<0,ΔG<0)with ITC determination indicated that the binding process between DPD and RbsB was exothermic and spontaneous,with hydrogen bonds and van der Waals forces as the main binding forces.Obviously,DPD can be more easily combined with RbsB in a dose-dependent manner,suggesting that RbsB was changed in the microenvironment of DPD when the DPD concentration was between 0.8-1.0mmolL−1 and reaching the maximum binding amount.According to molecular docking,3 hydrophobic residues involved in DPD and RbsB protein stable binding were be found,and also hydrogen bonding plays a key role in the formation of the new complex.RbsB efficiently inhibited V.harveyi bioluminescence induced by both S.agalactiae AI-2 and V.harveyi AI-2 in a dose-dependent manner.However,our results suggest that RbsB may play a role in the response of S.agalactiace to AI-2.展开更多
Isothermal titration calorimetry （ITC） was used to study interactions between water vapour and the surface of thermally converted sodium bicarbonate （NaHCO3）. The decarboxyla- tion degree of the samples was varied...Isothermal titration calorimetry （ITC） was used to study interactions between water vapour and the surface of thermally converted sodium bicarbonate （NaHCO3）. The decarboxyla- tion degree of the samples was varied from 3% to 35% and the humidity range was 54-100%. The obtained enthalpy values were all exothermic and showed a positive linear correlation with decarboxylation degrees for each humidity studied. The critical humidity, 75% （RHo）, was determined as the inflection point on a plot of the mean-AH kJ/mole Na2CO3 against RH. Humidities above the critical humidity lead to complete surface dissolution. The water uptake （m） was determined after each calorimetric experiment, complementing the enthalpy data. A mechanism of water vapour interaction with decarboxylated samples, including the formation of trona and Wegscheider＇s salt on the bicarbonate surface is proposed for humidities below RHo.展开更多
This study was aimed at gaining further insight on the role of hydration in adsorption processes of biopolymer/adsorbate systems using complementary methods (electromagnetic interference (EMI) shielding, calorimetry, ...This study was aimed at gaining further insight on the role of hydration in adsorption processes of biopolymer/adsorbate systems using complementary methods (electromagnetic interference (EMI) shielding, calorimetry, and solvent/vapour adsorption isotherms). Cellulose and starch-based materials were used as the adsorbents, whereas water (liquid and vapour), ethanol and p-nitrophenol (PNP) in aqueous solution were the adsorbate systems. The biopolymer/water systems had higher uptake capacity overall, where starch materials showed higher uptake capacity than cellulose among the various solvents. The secondary and tertiary structure of the biopolymers was a key factor affecting their uptake capacity, as evidenced by the enhanced adsorption properties of starch over cellulose, along with higher uptake of amylose (AM) versus amylopectin (AP) in starch biopolymers. EMI results also confirmed that AM starch had higher adsorption toward water than ethanol. The textural properties and surface chemistry of the biopolymers were probed using dye adsorption (PNP at pH 8.5) in aqueous solution that showed parallel trends with water vapour adsorption isotherms. Isothermal Titration Calorimetry (ITC) revealed that the heat of adsorption in AP differed from that of AM since the biopolymer tertiary structure governs the accessibility of biopolymer adsorption sites. The role of branching in AP and amorphous domains in AM/AP composites are inferred to play a key role in hydration-driven allosterism known for such biopolymer/water vapour adsorption processes.展开更多
Biophysical characterization has become an important experimental approach to understanding new details of the important structure-function relationship of
基金This study was supported by the National Natural Sci-ence Foundation of China(Nos.31702386,31660251,31860245 and 31960203)the International Cooperation Science&Technology Planning Project of Guangdong Province of China(No.2017A050501037)+2 种基金the Natural Science Foundation of Guangxi Province(Nos.2018 GXNSFAA281019,2017GXNSFAA198010)the Central Government Directs Special Funds for Local Science and Technology Development Projects(No.ZY1949015)E.Wangkahart was supported financially by the Ministry of Science and Technology of Thailand and Mahasarakham University.
文摘Understanding aquatic pathogen in sediments or aquacultural water is crucial to protect public health from soilborne and waterborne diseases.Quorum sensing(QS)was increasingly reported in biological wastewater treatment processes because of their inherent roles in biofilm development,bacterial aggregation and so on.The widely QS signals was Antoinducer-2(AI-2),primarily involved to allow the possibility of interspecies communication.However,the cellular components that mediate the response of Streptococcus agalactiae to AI-2 have not been fully characterized.Analysis of the complete genome sequence of S.agalactiae indi-cated that its RbsB protein has similarity to Escherichia coli LsrB and Aggregatibacter actinomycetemcomitans RbsB proteins that bind AI-2.We hypothesized that RbsB protein mediates quorum sensing signal uptake via interaction with AI-2.To evaluate the regulatory effect of RbsB on QS system,the recombinant plasmid pGEX-6p-1-RbsB was constructed and RbsB protein was purified with GST-tag.To further elucidate the role of RbsB protein binding to DPD(AI-2 precursor dihydroxypentanedione),the systemati-cally throughput circular dichroism(CD)spectroscopy,isothermal titration calorimetry200(ITC200)and molecular docking methods were employed.The high expression of soluble RbsB protein with molecular weight of 33 kDa was obtained.The thermodynamics results(ΔH<0,ΔS<0,ΔG<0)with ITC determination indicated that the binding process between DPD and RbsB was exothermic and spontaneous,with hydrogen bonds and van der Waals forces as the main binding forces.Obviously,DPD can be more easily combined with RbsB in a dose-dependent manner,suggesting that RbsB was changed in the microenvironment of DPD when the DPD concentration was between 0.8-1.0mmolL−1 and reaching the maximum binding amount.According to molecular docking,3 hydrophobic residues involved in DPD and RbsB protein stable binding were be found,and also hydrogen bonding plays a key role in the formation of the new complex.RbsB efficiently inhibited V.harveyi bioluminescence induced by both S.agalactiae AI-2 and V.harveyi AI-2 in a dose-dependent manner.However,our results suggest that RbsB may play a role in the response of S.agalactiace to AI-2.
文摘Isothermal titration calorimetry （ITC） was used to study interactions between water vapour and the surface of thermally converted sodium bicarbonate （NaHCO3）. The decarboxyla- tion degree of the samples was varied from 3% to 35% and the humidity range was 54-100%. The obtained enthalpy values were all exothermic and showed a positive linear correlation with decarboxylation degrees for each humidity studied. The critical humidity, 75% （RHo）, was determined as the inflection point on a plot of the mean-AH kJ/mole Na2CO3 against RH. Humidities above the critical humidity lead to complete surface dissolution. The water uptake （m） was determined after each calorimetric experiment, complementing the enthalpy data. A mechanism of water vapour interaction with decarboxylated samples, including the formation of trona and Wegscheider＇s salt on the bicarbonate surface is proposed for humidities below RHo.
文摘This study was aimed at gaining further insight on the role of hydration in adsorption processes of biopolymer/adsorbate systems using complementary methods (electromagnetic interference (EMI) shielding, calorimetry, and solvent/vapour adsorption isotherms). Cellulose and starch-based materials were used as the adsorbents, whereas water (liquid and vapour), ethanol and p-nitrophenol (PNP) in aqueous solution were the adsorbate systems. The biopolymer/water systems had higher uptake capacity overall, where starch materials showed higher uptake capacity than cellulose among the various solvents. The secondary and tertiary structure of the biopolymers was a key factor affecting their uptake capacity, as evidenced by the enhanced adsorption properties of starch over cellulose, along with higher uptake of amylose (AM) versus amylopectin (AP) in starch biopolymers. EMI results also confirmed that AM starch had higher adsorption toward water than ethanol. The textural properties and surface chemistry of the biopolymers were probed using dye adsorption (PNP at pH 8.5) in aqueous solution that showed parallel trends with water vapour adsorption isotherms. Isothermal Titration Calorimetry (ITC) revealed that the heat of adsorption in AP differed from that of AM since the biopolymer tertiary structure governs the accessibility of biopolymer adsorption sites. The role of branching in AP and amorphous domains in AM/AP composites are inferred to play a key role in hydration-driven allosterism known for such biopolymer/water vapour adsorption processes.
文摘Biophysical characterization has become an important experimental approach to understanding new details of the important structure-function relationship of
