A factorial study was conducted to determine the effects of salinity （constant （SO） and fluctuating （S10）） and isoenergetic diet of different protein/carbohydrate ratios （P/C： 4.1, 1.9, 1.0 and 0.6） on the su...A factorial study was conducted to determine the effects of salinity （constant （SO） and fluctuating （S10）） and isoenergetic diet of different protein/carbohydrate ratios （P/C： 4.1, 1.9, 1.0 and 0.6） on the survival and growth of juvenile Litopenaeus vannamei. The experiment lasted for 35 days. The results showed that I） At S0 and S10, specific growth rate and apparent digestibility coefficient exhibited decreasing trend, while food conversion ratio exhibited increasing trend, with decreasing P/C ratio. At SO, food consumption and specific growth rate of the shrimp fed 4.1 P/C were significantly higher than those fed 1.0 and 0.6 P/C, whereas at S10, no significant differences were detected among different treatments; II） At 4.1 and 1.9 P/C, food consumption and specific growth rate were higher in the shrimp maintained at SO as compared with those at S10, while at 1.0 and 0.6 P/C, an inverse trend was observed. Among four P/C ratios, food conversion ratio of the shrimp at SO was the lowest.展开更多
Proteomics was used to reveal the differential protein expression profiles of acute responses to copper sulfate exposure in larvae of Artemia sinica.Fourteen differentially displayed protein spots were detected and se...Proteomics was used to reveal the differential protein expression profiles of acute responses to copper sulfate exposure in larvae of Artemia sinica.Fourteen differentially displayed protein spots were detected and seven of them were identified.Three spots were up-expressed and identified:actin, heat shock protein 70,and chaperone subunit 1;three down-regulated proteins were identified:arginine kinase,elongation factor-2,and glycine-rich protein;and a newly expressed protein was identified as peroxiredoxin.The study indicates the involvement of all the differentially expressed proteins in the early responses of protein expression,and in the survival of A.sinica in the presence of copper and other heavy metals;the findings improve understanding of the organism’s adaptive responses and resistance.展开更多
The native protein structures in buffer solution are maintained by the electrostatic force as well as the hydrophobic force, salt ions play an important role in maintaining the protein native structures, and their eff...The native protein structures in buffer solution are maintained by the electrostatic force as well as the hydrophobic force, salt ions play an important role in maintaining the protein native structures, and their effect on the protein stability has attracted tremendous interests. Infrared spectroscopy has been generally used in molecular structure analysis due to its fingerprint resolution for different species including macromolecules as proteins. However spectral intensities have received much less attention than the vibrational frequencies. Here we report that the spectral intensities of protein amide I band, the finger prints for the protein secondary structures, are very sensitive to the local electric field known as Onsager reaction field caused by salt ions. IR absorbance thermal titrations have been conducted for a series of samples including simple water soluble amino acids, water soluble monomeric protein cytochrome c and dimeric protein DsbC and its single-site mutant G49R. We found that at lower temperature range （10-20℃）, there exists a thermal activated salting-in process, where the IR intensity increases with a rise in the temperature, corresponding to the ions binding of the hydrophobic surface of protein. This process is absent for the amino acids. When further raising the temperature, the IR intensity decreases, this is interpreted as the thermal activated breaking of the ion-protein surface binding. Applying Van＇t Hoff plot to the thermal titration curves, the thermodynamic parameters such as AH and AS for salting-in and ion unbinding processes can be derived for various protein secondary structural components, revealing quantitatively the extent of hydrophobic interaction as well as the strength of the ion-protein binding.展开更多
The expression of recombinant proteins in microorganism frequently leads to the formation of insoluble aggregates, inclusion bodies （IBs）. Thus, the additional in vitro protein refolding process is required to conve...The expression of recombinant proteins in microorganism frequently leads to the formation of insoluble aggregates, inclusion bodies （IBs）. Thus, the additional in vitro protein refolding process is required to convert inactive IBs into water-soluble active proteins. This study investigated the effect of sulfur residue and hydrophobicity of imidazolium-based room temperature ionic liquids （RTILs） on the refolding of lysozyme as a model protein in the batch dilution method which is the most commonly used refolding method. When lysozyme was refolded in the refolding buffer containing [BF4]-based RTILs with a systematic variety of alkyl chain on cations varying from two to eight, less hydrophobic imidazolium cations having shorter alkyl chains were effective to facilitate lysozyme refolding. Compared to the conventional refolding buffer, 2 times higher lysozyme re- folding yield was obtained in l-ethyl-3-methylimidazolium tetrafluoroborate （[EMIM][BF4]） containing refolding buffer. The refolding yield of lysozyme was even more increased by 2.5 times when 1-butyl-3-methylimidazolium methylsulfate （[BMIM][MS]） containing sulfur residue on anion was used. The sulfur residue in [BMIM][MS] is supposed to improve the refolding yield of lysozyme which has 4 intrarnolecular disulfide bonds. For dilution-based refolding of lysozyme, the opti- mum concentrations of RTILs in refolding buffer were found to be 1.0 M [EMIM][BF4] and 0.5 M [PMIM][MS], respectively. The optimum temperate for dilution-based refolding of lysozyme with RTILs was 4 ℃.展开更多
基金funded by National Key Project of Scientific and Technical Supporting Programs funded by Ministry of Science & Technology of China (Grant No. 2006BAD09A01, 2006BAD09A06)Jiangsu Provincial Science Foundation for Talent Youths (Grant No. 2006548) Open Fund of Jiangsu Key Laboratory of Marine Biotechnology (Grant No. 2006HS017)
文摘A factorial study was conducted to determine the effects of salinity （constant （SO） and fluctuating （S10）） and isoenergetic diet of different protein/carbohydrate ratios （P/C： 4.1, 1.9, 1.0 and 0.6） on the survival and growth of juvenile Litopenaeus vannamei. The experiment lasted for 35 days. The results showed that I） At S0 and S10, specific growth rate and apparent digestibility coefficient exhibited decreasing trend, while food conversion ratio exhibited increasing trend, with decreasing P/C ratio. At SO, food consumption and specific growth rate of the shrimp fed 4.1 P/C were significantly higher than those fed 1.0 and 0.6 P/C, whereas at S10, no significant differences were detected among different treatments; II） At 4.1 and 1.9 P/C, food consumption and specific growth rate were higher in the shrimp maintained at SO as compared with those at S10, while at 1.0 and 0.6 P/C, an inverse trend was observed. Among four P/C ratios, food conversion ratio of the shrimp at SO was the lowest.
基金Supported by the Natural Science Foundation of China(No.20060109Z4016)the National Basic Research Development Program of China(No.2006CB101804)the Natural Science Foundation of Shandong Provincefor the excellent young researchers(No.2006BSA02004)
文摘Proteomics was used to reveal the differential protein expression profiles of acute responses to copper sulfate exposure in larvae of Artemia sinica.Fourteen differentially displayed protein spots were detected and seven of them were identified.Three spots were up-expressed and identified:actin, heat shock protein 70,and chaperone subunit 1;three down-regulated proteins were identified:arginine kinase,elongation factor-2,and glycine-rich protein;and a newly expressed protein was identified as peroxiredoxin.The study indicates the involvement of all the differentially expressed proteins in the early responses of protein expression,and in the survival of A.sinica in the presence of copper and other heavy metals;the findings improve understanding of the organism’s adaptive responses and resistance.
基金This work was supported by the National Natural Science Foundation of China （No.20373088）, the Program for Innovation Group （No.60321002）, the Innovative Project of Chinese Academy of Sciences （No.KJCX2-SW-w29）, and the National Key Project for Basic Research No.2006CB910302）. We thank Prof. Chih-chen Wang and Dr. Hui-min Ke in the Institute of Biophysics, Chinese Academy of Science, for the preparation of samples DsbC and G49R. We also thank Prof. Xiang-gang Qiu in the Institute of physics, Chinese Academy of Sciences, for help in FTIR measurement.
文摘The native protein structures in buffer solution are maintained by the electrostatic force as well as the hydrophobic force, salt ions play an important role in maintaining the protein native structures, and their effect on the protein stability has attracted tremendous interests. Infrared spectroscopy has been generally used in molecular structure analysis due to its fingerprint resolution for different species including macromolecules as proteins. However spectral intensities have received much less attention than the vibrational frequencies. Here we report that the spectral intensities of protein amide I band, the finger prints for the protein secondary structures, are very sensitive to the local electric field known as Onsager reaction field caused by salt ions. IR absorbance thermal titrations have been conducted for a series of samples including simple water soluble amino acids, water soluble monomeric protein cytochrome c and dimeric protein DsbC and its single-site mutant G49R. We found that at lower temperature range （10-20℃）, there exists a thermal activated salting-in process, where the IR intensity increases with a rise in the temperature, corresponding to the ions binding of the hydrophobic surface of protein. This process is absent for the amino acids. When further raising the temperature, the IR intensity decreases, this is interpreted as the thermal activated breaking of the ion-protein surface binding. Applying Van＇t Hoff plot to the thermal titration curves, the thermodynamic parameters such as AH and AS for salting-in and ion unbinding processes can be derived for various protein secondary structural components, revealing quantitatively the extent of hydrophobic interaction as well as the strength of the ion-protein binding.
基金supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0013308)
文摘The expression of recombinant proteins in microorganism frequently leads to the formation of insoluble aggregates, inclusion bodies （IBs）. Thus, the additional in vitro protein refolding process is required to convert inactive IBs into water-soluble active proteins. This study investigated the effect of sulfur residue and hydrophobicity of imidazolium-based room temperature ionic liquids （RTILs） on the refolding of lysozyme as a model protein in the batch dilution method which is the most commonly used refolding method. When lysozyme was refolded in the refolding buffer containing [BF4]-based RTILs with a systematic variety of alkyl chain on cations varying from two to eight, less hydrophobic imidazolium cations having shorter alkyl chains were effective to facilitate lysozyme refolding. Compared to the conventional refolding buffer, 2 times higher lysozyme re- folding yield was obtained in l-ethyl-3-methylimidazolium tetrafluoroborate （[EMIM][BF4]） containing refolding buffer. The refolding yield of lysozyme was even more increased by 2.5 times when 1-butyl-3-methylimidazolium methylsulfate （[BMIM][MS]） containing sulfur residue on anion was used. The sulfur residue in [BMIM][MS] is supposed to improve the refolding yield of lysozyme which has 4 intrarnolecular disulfide bonds. For dilution-based refolding of lysozyme, the opti- mum concentrations of RTILs in refolding buffer were found to be 1.0 M [EMIM][BF4] and 0.5 M [PMIM][MS], respectively. The optimum temperate for dilution-based refolding of lysozyme with RTILs was 4 ℃.
