In this study,circular dichroism(CD)and molecular dynamics(MD)simulation were used to investigate the thermal unfolding pathway of staphylococcal enterotoxin B(SEB)at temperatures of 298–371 and 298–500 K,and the re...In this study,circular dichroism(CD)and molecular dynamics(MD)simulation were used to investigate the thermal unfolding pathway of staphylococcal enterotoxin B(SEB)at temperatures of 298–371 and 298–500 K,and the relationship between the experimental and simulation results were explored.Our computational findings on the secondary structure of SEB showed that at room temperature,the CD spectroscopic results were highly consistent with the MD results.Moreover,under heating conditions,the changing trends of helix,sheet and random coil obtained by CD spectral fitting were highly consistent with those obtained by MD.In order to gain a deeper understanding of the thermal stability mechanism of SEB,the MD trajectories were analyzed in terms of root mean square deviation(RMSD),secondary structure assignment(SSA),radius of gyration(R_(g)),free energy surfaces(FES),solvent-accessible surface area(SASA),hydrogen bonds and salt bridges.The results showed that at low heating temperature,domain Ⅰ without loops(omitting the mobile loop region)mainly relied on hydrophobic interaction to maintain its thermal stability,whereas the thermal stability of domain Ⅱ was mainly controlled by salt bridges and hydrogen bonds.Under high heating temperature conditions,the hydrophobic interactions in domain Ⅰ without loops were destroyed and the secondary structure was almost completely lost,while domain Ⅱ could still rely on salt bridges as molecular staples to barely maintain the stability of the secondary structure.These results help us to understand the thermodynamic and kinetic mechanisms that maintain the thermal stability of SEB at the molecular level,and provide a direction for establishing safer and more effective food sterilization processes.展开更多
Animal models are essential for the development of new anti-infectious drugs.Although some bacterial infection models have been established in rodents,small primate models are rare.Here,we report on two bacterial infe...Animal models are essential for the development of new anti-infectious drugs.Although some bacterial infection models have been established in rodents,small primate models are rare.Here,we report on two bacterial infection models established in tree shrew（Tupaia belangeri chinensis）.A burnt skin infection model was induced by dropping 5×106 CFU of Staphylococcus aureus on the surface of a wound after a third degree burn.This dose of S.aureus caused persistent infection for 7 days and obvious inflammatory response was observed 4 days after inoculation.A Dacron graft infection model,2×106 CFU of Pseudomonas aeruginosa also caused persistent infection for 6 days,with large amounts of pus observed 3 days after inoculation.These models were used to evaluate the efficacy of levofloxacin（LEV） and cefoperazone（CPZ）,which reduced the viable bacteria in skin to 4log10 and 5log10 CFU/100 mg tissue,respectively.The number of bacteria in graft was significantly reduced by 4log10 CFU/mL treatment compared to the untreated group（P0.05）.These results suggest that two bacterial infection models were successfully established in tree shrew using P.aeruginosa and S.aureus.In addition,tree shrew was susceptible to P.aeruginosa and S.aureus,thus making it an ideal bacterial infection animal model for the evaluation of new antimicrobials.展开更多
[Objective] To produce drug resistance, seek non-toxic environmental so as to change the current biological drugs that did not excessive use of antibiotics. [Methods] A strain of Bacillus was purified and isolated fro...[Objective] To produce drug resistance, seek non-toxic environmental so as to change the current biological drugs that did not excessive use of antibiotics. [Methods] A strain of Bacillus was purified and isolated from fresh and healthy in- testines of grass carps. Biochemical identification was carried out by conventional bacterial biochemical test method. Two pairs of primers were designed, 16S rRNA detection and sequencing analysis were carried out. Drug sensitive test was carried out by agar diffusion method. In vitro inhibition test on Staphylococcus aureus was carried out by Oxford cup method. [Results] The isolated bacterium had basically the same biochemical characters as Bacillus subtilis; and the homology reached 100%. Thus, the isolated bacterium was identified to be Bacillus subtilis. It was insensitive to amoxicillin, ampicillin, penicillin G and so on, but sensitive to amikacin, cefalexin, ciprofloxacin and cefradine. The inhibitory effects of Bacillus subtilis on Staphylococ- cus aureus were significant. The minimum inhibitory concentration （MIC） was 2.8×10^8×2^-5/ml and minimum bactericidal concentration （MBC） was 2.8×10^8×2^-2/ml. [Conclusions] The isolated Bacillus subtilis could be used to prevent and control diseases caused by Staphylococcus aureus, and reduce the abuse of antibiotics.展开更多
文摘In this study,circular dichroism(CD)and molecular dynamics(MD)simulation were used to investigate the thermal unfolding pathway of staphylococcal enterotoxin B(SEB)at temperatures of 298–371 and 298–500 K,and the relationship between the experimental and simulation results were explored.Our computational findings on the secondary structure of SEB showed that at room temperature,the CD spectroscopic results were highly consistent with the MD results.Moreover,under heating conditions,the changing trends of helix,sheet and random coil obtained by CD spectral fitting were highly consistent with those obtained by MD.In order to gain a deeper understanding of the thermal stability mechanism of SEB,the MD trajectories were analyzed in terms of root mean square deviation(RMSD),secondary structure assignment(SSA),radius of gyration(R_(g)),free energy surfaces(FES),solvent-accessible surface area(SASA),hydrogen bonds and salt bridges.The results showed that at low heating temperature,domain Ⅰ without loops(omitting the mobile loop region)mainly relied on hydrophobic interaction to maintain its thermal stability,whereas the thermal stability of domain Ⅱ was mainly controlled by salt bridges and hydrogen bonds.Under high heating temperature conditions,the hydrophobic interactions in domain Ⅰ without loops were destroyed and the secondary structure was almost completely lost,while domain Ⅱ could still rely on salt bridges as molecular staples to barely maintain the stability of the secondary structure.These results help us to understand the thermodynamic and kinetic mechanisms that maintain the thermal stability of SEB at the molecular level,and provide a direction for establishing safer and more effective food sterilization processes.
基金financially supported by the Project from the Chinese Academy of Sciences (KSCX2-EW-R-11)the Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences&Yunnan Province (KSCX2-EW-J-23)Science and Technology Department of Yunnan Province (2011C1139)
文摘Animal models are essential for the development of new anti-infectious drugs.Although some bacterial infection models have been established in rodents,small primate models are rare.Here,we report on two bacterial infection models established in tree shrew（Tupaia belangeri chinensis）.A burnt skin infection model was induced by dropping 5×106 CFU of Staphylococcus aureus on the surface of a wound after a third degree burn.This dose of S.aureus caused persistent infection for 7 days and obvious inflammatory response was observed 4 days after inoculation.A Dacron graft infection model,2×106 CFU of Pseudomonas aeruginosa also caused persistent infection for 6 days,with large amounts of pus observed 3 days after inoculation.These models were used to evaluate the efficacy of levofloxacin（LEV） and cefoperazone（CPZ）,which reduced the viable bacteria in skin to 4log10 and 5log10 CFU/100 mg tissue,respectively.The number of bacteria in graft was significantly reduced by 4log10 CFU/mL treatment compared to the untreated group（P0.05）.These results suggest that two bacterial infection models were successfully established in tree shrew using P.aeruginosa and S.aureus.In addition,tree shrew was susceptible to P.aeruginosa and S.aureus,thus making it an ideal bacterial infection animal model for the evaluation of new antimicrobials.
基金Supported by the Cooperation Subject(09003699)the Project of Jiangxi Education Department(GJJ12237)the Project of Science and Technology Department of Jiangxi(20122BBF60082)~~
文摘[Objective] To produce drug resistance, seek non-toxic environmental so as to change the current biological drugs that did not excessive use of antibiotics. [Methods] A strain of Bacillus was purified and isolated from fresh and healthy in- testines of grass carps. Biochemical identification was carried out by conventional bacterial biochemical test method. Two pairs of primers were designed, 16S rRNA detection and sequencing analysis were carried out. Drug sensitive test was carried out by agar diffusion method. In vitro inhibition test on Staphylococcus aureus was carried out by Oxford cup method. [Results] The isolated bacterium had basically the same biochemical characters as Bacillus subtilis; and the homology reached 100%. Thus, the isolated bacterium was identified to be Bacillus subtilis. It was insensitive to amoxicillin, ampicillin, penicillin G and so on, but sensitive to amikacin, cefalexin, ciprofloxacin and cefradine. The inhibitory effects of Bacillus subtilis on Staphylococ- cus aureus were significant. The minimum inhibitory concentration （MIC） was 2.8×10^8×2^-5/ml and minimum bactericidal concentration （MBC） was 2.8×10^8×2^-2/ml. [Conclusions] The isolated Bacillus subtilis could be used to prevent and control diseases caused by Staphylococcus aureus, and reduce the abuse of antibiotics.
