ESR技术应用于逆胶束酶体系性能的研究
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摘要
采用鼠李糖脂构建逆胶束体系,并利用电子自旋共振(ESR)技术研究了鼠李糖脂逆胶束及鼠李糖脂逆胶束酶体系的性能.采用ESR光谱技术计算得出逆胶束体系中超精细分裂常数变化,研究表明鼠李糖脂在正己烷中的临界胶度为0.07mmol/L.通过分析ESR光谱中旋转相关时间的变化,探究电子自旋探针在逆胶束中运动受阻和翻转所需时间的情况同时,ESR光谱的峰值体现了样品中自由基含量的多少.通过对比逆胶束体系、逆胶束酶体系以及逆胶束酶-苯酚体系的ESR光谱,结果表明鼠李糖脂逆胶束-苯酚体系的自由基最多.通过研究对16-氮氧自由基硬脂酸对两个体系作用,推测出电子自旋探针自由基团定位于逆胶束水核中,并且推断电子自旋探针位于水核的边缘区域,即结合水水层.该研究为逆胶束酶体系的应用提供了坚实的理论基础.
In this study, a reverse micelle system was constructed using rhamnolipid, and the properties of rhamnolipid reverse micellesand rhamnolipid reverse micelle enzyme system were studied by electron spin resonance(ESR) technique. ESR spectroscopy was usedto calculate the performance. The ultrafine splitting constant in the reverse micelle system was obtained. The study showed that thecriticality of rhamnolipid in n-hexane was 0.07 mmol/L. By analyzing the change of rotation correlation time in ESR spectrum, theelectron spin probe was explored. At the same time, the peak of the ESR spectrum reflects the amount of free radicals in the sample. Bycomparing the reverse micelle system, the reverse micelle enzyme system and the reverse micelle enzyme-phenol system ESR spectrashowed that the rhamnolipid reversed micelle-phenol system had the most free radicals. By studying the effect of 16-nitroxyl radicalstearic acid on two systems, it was speculated that the electron spin probe free radicals were localized. In the reverse micelle water core,and inferred that the electron spin probe was located in the edge region of the water core, that was, the water-water layer was combined.This study provided a solid theoretical basis for the application of the reverse micelle enzyme system.
In this study, a reverse micelle system was constructed using rhamnolipid, and the properties of rhamnolipid reverse micellesand rhamnolipid reverse micelle enzyme system were studied by electron spin resonance(ESR) technique. ESR spectroscopy was usedto calculate the performance. The ultrafine splitting constant in the reverse micelle system was obtained. The study showed that thecriticality of rhamnolipid in n-hexane was 0.07 mmol/L. By analyzing the change of rotation correlation time in ESR spectrum, theelectron spin probe was explored. At the same time, the peak of the ESR spectrum reflects the amount of free radicals in the sample. Bycomparing the reverse micelle system, the reverse micelle enzyme system and the reverse micelle enzyme-phenol system ESR spectrashowed that the rhamnolipid reversed micelle-phenol system had the most free radicals. By studying the effect of 16-nitroxyl radicalstearic acid on two systems, it was speculated that the electron spin probe free radicals were localized. In the reverse micelle water core,and inferred that the electron spin probe was located in the edge region of the water core, that was, the water-water layer was combined.This study provided a solid theoretical basis for the application of the reverse micelle enzyme system.
引文
[1]Li F,Li G Z,Wang H Q,et al.Studies on cetyltrimethylammonium bromide(CTAB)micellar solution and CTAB reversed microemulsion by ESR and 2H NMR[J].Colloids and Surfaces A,1997,127:89-96.
[2]Samanta A,Paul B K,Guchhait N.Spectroscopic probe analysis for exploring probe-protein interaction:a mapping of native,unfolding and refolding of protein bovine serum albumin by extrinsic fluorescence probe[J].Biophysical Chemistry,2011,156(2):128-139.
[3]Hisashi Y,Shunsuke K.Spectral Simulation Study of the Positional Exchange of a Spin Probe in an Aerosol OT Reversed Micelle[J].Journal of Colloid and Interface Science,1983,95:240-246.
[4]Giuseppe O,Ado S.IR Investigations of Water Structure in Aerosol OT Reverse Micellar Aggregates[J].Journal of Physical Chemistry,1993,97:5430-5434.
[5]谈菲,刘智峰,钟华,等.低浓度鼠李糖脂单糖脂在近中性溶液中的聚集行为[J].中国环境科学,2014,34(11):2912-2916.Tan F,Liu Z F,Zhong H,et al.Aggregation of monorhamnolipid with concentrations near CMC in neutral solution[J].China Environmental Science,2014,34(11):2912-2916.
[6]Haering G,Luisi P L,Hauser H.Characterization by electron spin resonance of reversed micelles consisting of the ternary system AOT-Isooctane-Water[J].Journal of Physical Chemistry,1988,92:3574-3581.
[7]Hisashi Y.Exchange of the position of a spin probe in an aerosol OTreversed micelle[J].Journal of Colloid and Interface Science,1983,95:81-86.
[8]王英彦,李小洁,贾文英,等.环境诱变剂和人血红细胞膜类脂相互作用的ESR波谱研究[J].中国环境科学,1989,(4):256-260.Wang Y Y,Li X J,Jia W Y,et al.ESR study on the interaction of environmental mutagens and human erythrocyte membrane[J].China Environmental Science,1989,(4):256-260.
[9]Tsai,Chia Jung.Spin-Label ESR study of the protein domain motion and stability in the presence of crowding effects[J].Biophysical Journal,2014,106(2):665-665.
[10]Nicholas J,Lei X G.Spectroscopic probe analysis of proteinsurfactant interactions:The BSA/SDS system[J].Langmuir,1995,11:2525-2533.
[11]Szajdzinska-Pietek E,Maldonado R,Kevan L.Effect of 1-butanol on the surface structure of ionic micelles and location of its solubilization site:Electron spin echo modulation study with x-doxylstearic acid spin probes[J].Journal of Colloid and Interface Science,1986,110:514-519.
[12]Szajdzinska-Pietek E,Maldonado R,Kevan L.Effect of 1-butanol on the surface structure of ionic micelles and location of its solubilization site:Electron spin echo modulation study with x-doxylstearic acid spin probes[J].Journal of Colloid and Interface Science,1986,110:514-519.
[13]Szajdzinska-Pietek E,Maldonado R,Kevan L,et al.Electron Spin-Echo modulation studies of doxylstearic acid spin probes in sodium and tetramethylammonium dodecyl sulfate micelles:Interaction of the spin probe with D20 and with deuterated terminal methyl groups in the surfactant molecules[J].Journal of the American Chemical Society,1985,107:784-788.
[14]Baglioni P,Rivara-Minten E,Kevan L.Electron spin resonance and electron spin echo modulation of n-doxylstearic acid and N,N,N/,N/-Tetramethylbenzldine photoionization in sodium versus lithium dodecyl sulfate m L cellar solutions:Effect of 15-Crown-5 and18-Crown-6Ether addition[J].Journal of Physical Chemistry,1988,92:4726-4130.
[15]Baglioni Piero,Bongiovanni R,Rivara-Minten E,et al.Electron spin-echo modulation of x-doxylstearic acid in nonionic surfactants[J].Journal of Physical Chemistry,1989,93:5574-5578.
[16]Zhao J X,Deng S J,Liu J Y,et al.Fourier transform infrared investigation on the state of water in reverse micelles of quaternary ammonium gemini surfactants C12-s-C12-2Br in n-heptane[J].Journal of Colloid and Interface Science,2007,311:237-242.
[17]Guo F Q,Li H F,Zhang Z F,et al.Reversed micelle formation in a model liquid-liquid extraction system[J].Journal of Colloid and Interface Science,2008,322:605-610
[2]Samanta A,Paul B K,Guchhait N.Spectroscopic probe analysis for exploring probe-protein interaction:a mapping of native,unfolding and refolding of protein bovine serum albumin by extrinsic fluorescence probe[J].Biophysical Chemistry,2011,156(2):128-139.
[3]Hisashi Y,Shunsuke K.Spectral Simulation Study of the Positional Exchange of a Spin Probe in an Aerosol OT Reversed Micelle[J].Journal of Colloid and Interface Science,1983,95:240-246.
[4]Giuseppe O,Ado S.IR Investigations of Water Structure in Aerosol OT Reverse Micellar Aggregates[J].Journal of Physical Chemistry,1993,97:5430-5434.
[5]谈菲,刘智峰,钟华,等.低浓度鼠李糖脂单糖脂在近中性溶液中的聚集行为[J].中国环境科学,2014,34(11):2912-2916.Tan F,Liu Z F,Zhong H,et al.Aggregation of monorhamnolipid with concentrations near CMC in neutral solution[J].China Environmental Science,2014,34(11):2912-2916.
[6]Haering G,Luisi P L,Hauser H.Characterization by electron spin resonance of reversed micelles consisting of the ternary system AOT-Isooctane-Water[J].Journal of Physical Chemistry,1988,92:3574-3581.
[7]Hisashi Y.Exchange of the position of a spin probe in an aerosol OTreversed micelle[J].Journal of Colloid and Interface Science,1983,95:81-86.
[8]王英彦,李小洁,贾文英,等.环境诱变剂和人血红细胞膜类脂相互作用的ESR波谱研究[J].中国环境科学,1989,(4):256-260.Wang Y Y,Li X J,Jia W Y,et al.ESR study on the interaction of environmental mutagens and human erythrocyte membrane[J].China Environmental Science,1989,(4):256-260.
[9]Tsai,Chia Jung.Spin-Label ESR study of the protein domain motion and stability in the presence of crowding effects[J].Biophysical Journal,2014,106(2):665-665.
[10]Nicholas J,Lei X G.Spectroscopic probe analysis of proteinsurfactant interactions:The BSA/SDS system[J].Langmuir,1995,11:2525-2533.
[11]Szajdzinska-Pietek E,Maldonado R,Kevan L.Effect of 1-butanol on the surface structure of ionic micelles and location of its solubilization site:Electron spin echo modulation study with x-doxylstearic acid spin probes[J].Journal of Colloid and Interface Science,1986,110:514-519.
[12]Szajdzinska-Pietek E,Maldonado R,Kevan L.Effect of 1-butanol on the surface structure of ionic micelles and location of its solubilization site:Electron spin echo modulation study with x-doxylstearic acid spin probes[J].Journal of Colloid and Interface Science,1986,110:514-519.
[13]Szajdzinska-Pietek E,Maldonado R,Kevan L,et al.Electron Spin-Echo modulation studies of doxylstearic acid spin probes in sodium and tetramethylammonium dodecyl sulfate micelles:Interaction of the spin probe with D20 and with deuterated terminal methyl groups in the surfactant molecules[J].Journal of the American Chemical Society,1985,107:784-788.
[14]Baglioni P,Rivara-Minten E,Kevan L.Electron spin resonance and electron spin echo modulation of n-doxylstearic acid and N,N,N/,N/-Tetramethylbenzldine photoionization in sodium versus lithium dodecyl sulfate m L cellar solutions:Effect of 15-Crown-5 and18-Crown-6Ether addition[J].Journal of Physical Chemistry,1988,92:4726-4130.
[15]Baglioni Piero,Bongiovanni R,Rivara-Minten E,et al.Electron spin-echo modulation of x-doxylstearic acid in nonionic surfactants[J].Journal of Physical Chemistry,1989,93:5574-5578.
[16]Zhao J X,Deng S J,Liu J Y,et al.Fourier transform infrared investigation on the state of water in reverse micelles of quaternary ammonium gemini surfactants C12-s-C12-2Br in n-heptane[J].Journal of Colloid and Interface Science,2007,311:237-242.
[17]Guo F Q,Li H F,Zhang Z F,et al.Reversed micelle formation in a model liquid-liquid extraction system[J].Journal of Colloid and Interface Science,2008,322:605-610
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