离子液体—水体系中蛋白质晶体的生长研究
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摘要
蛋白质晶体学是一门研究蛋白质结构与功能的边缘学科。随着生物学领域对精确的生物大分子结构需求的日益增加,尤其是利用单晶X射线衍射法得到的三维结构,使得研究生物大分子晶体的生长越来越受到重视。然而,生物大分子的结晶相当困难,从某种意义上讲这是生物大分子结构测定的限制因素。
离子液体具有独特的结构和物理化学性质,已经成为目前化学界的热点课题之一,而且在生命科学领域也开始得到应用,尤其是作为生物催化反应的溶剂或共溶剂的研究更是备受关注,近年来,离子液体也开始应用于蛋白质结晶方面。
本论文将一种新体系——离子液体-水用于蛋白质的结晶。在该体系中,实现了蛋白质晶体在离子液体中的可控生长,培养了符合X射线衍射条件的蛋白质单晶,对离子液体在蛋白质结晶过程中的作用机理进行了探讨,并剖析了离子液体含量与蛋白质晶体的晶型、形貌和品质间的关系。此外,从动力学角度系统地分析了离子液体在蛋白质结晶过程中对晶体生长速率的控制作用,进而优化结晶条件,改善晶体质量。
采用一步法和两步法分别合成了离子液体[BMIM]Cl(1-丁基-3-甲基咪唑氯化物)、[BMIM]BF_4(1-丁基-3-甲基咪唑四氟硼酸盐)和[BMIM]PF_6(1-丁基-3-甲基咪唑六氟磷酸盐),借助红外光谱分析表征了其化学结构。其次,为了进一步研究离子液体对蛋白质晶体生长过程的影响机制,采用BCA法通过紫外—可见分光光度计测定蛋白质在离子液体—水混合溶液中的溶解度,结果表明,离子液体在体系中起到了类似无机盐类沉淀剂的作用,促进了蛋白质分子从溶液中的析出。通过测定[BMIM]BF_4-乙酸乙酸钠-NaCl离子液体双水相体系的三元体系相图,研究了培养溶液中各组分间的溶解度关系,并将其用于蛋白质晶体的培养,以控制了溶液的局部过饱和度。
采用悬滴法和批量法两种结晶方法,在离子液体-水体系中实现了对溶菌酶晶体的可控生长,得到了空间群为P4_32_12的四方晶系晶体,并对其结构进行了模型解析工作。考察了沉淀剂种类、溶液过饱和度、离子液体的含量、结晶温度等实验参数的改变对溶菌酶晶型、晶体形貌和品质的影响。研究表明,[BMIM]BF_4更适于溶菌酶晶体的培养,在该体系中,[BMIM]BF_4与NaCl一起作为联合沉淀剂使用,晶体尺寸明显增大,结晶速率得以控制,晶体质量有所改善。另外,随着[BMIM]BF_4含量的增加,晶体形貌逐渐由块状变为棒状,且长径比也随之增大。
采用悬滴结晶法,在[BMIM]BF_4-水体系中实现了对索马甜蛋白单晶体的可控生长,得到了空间群为P4_12_12的四方晶系晶体,并解析了其初始模型结构。在该体系中,[BMIM]BF_4被单独的作为沉淀剂使用,并且研究了结晶温度、离子液体[BMIM]BF_4的含量等培养条件对索马甜蛋白晶型和品质的影响。
离子液体作为一种具有特殊结构和功能的溶剂,在蛋白质结晶的过程中具有与无机盐类沉淀剂相类似的作用。同时形成双水相体系降低溶液的局部过饱和度,发挥了模板剂、联合沉淀剂和添加剂的多重作用,有效的控制晶核的生成数量,提高单晶出现的几率。可以通过调节离子液体的含量来控制蛋白质晶体的生长速率、大小和质量。
Protein crystallography is an interdisciplinary subject in the research of the structure and function of the protein. With the increasing demands of precise biomolecular 3-D structures, especially protein structures by X-ray diffraction, people pay more and more attention to the study of protein crystal growth. However, the crystallization of protein is very difficult and become a bottleneck in structural genomics, to some extent.
Ionic liquids (ILs) with unique structure and outstanding physical and chemical properties have become the focus in the chemistry field. And it is also used in the field of life science, especially in the biocatalyst reaction as solvent or cosolvent. Recently, ionic liquids are also applied in protein crystallization.
In this work, a new system of ionic liquid-water was tested for the protein crystallization. The procedure of crystallization could be controllable conducted in this system. The mechanisms for formations of controlled growth and the relationship among polymorph, morphology, quality and ionic liquid content were investigated. Moreover, the limitation effect of ionic liquid on the rate of crystal growth was disscussed from the view of kinetics.
Ionic liquids of [BMIM]Cl, [BMIM]BF_4 and [BMIM]PF_6 were prepared through Direct Method and Two-step Method, respectively. And the chemical structures of the final products were characterized by Infrared spectra. For the further studies on the effects of the ILs on the crystal growth, the solubility of protein was mensurated with ultraviolet-visible light spectrophotometer by the way of BCA method. It was found that ionic liquid had similar effect as precipitants or additions, which was favourable for the single crystals. On the other hand, in order to understand their dissolved relationship among each components in the solution, the equilibrium solubility of a ternary system of [BMIM]BF_4, NaCl and buffer as well as the data of liquid-liquid equilibrium of that system are determined at 25℃and atmospheric pressure. It was used for the protein crystallization, and played a role in the conrol of local supersaturation.
Two crystallization methods (hanging drop and sitting drop crystallization) were used for the lysozyme crystallization in the ionic liquids with different buffer/IL proportions. Lysozyme crystal belonged to the tetragonal space group P4_32_12 and its 3-D model was analyzed. The influence of growth conditions such as the kind of precipitants, solution supersaturation, ionic liquids content and temperature on polymorph, morphology, quality of crystals was discussed. It showed that [BMIM]BF_4 was more suitable for lysozyme crystallization than other ILs. [BMIM]BF_4 and NaCl were used as union-precipitants in the system of [BMIM]BF_4-water. More lager single crystals with controllable morphologies could be obtained due to the manageable crystal growth velocity. In addition, the morphology of lysozyme crystals were changed from bolck to rod and the L/D ratio increased with the proportion of IL.
Futhermore, thaumatin crystal, which was belonged to the tetragonal space group P4_12_12, could be obtained with hanging drop crystallization. [BMIM]BF_4 was used as a precipitant, and the effect of ionic liquids content, temperature on polymorph and quality of crystals was investigated.
On a whole, ionic liquid was a solvent with special structure and function. More importantly, aqueous two-phase system was appeared with diffusion of water molecules. As a new system, it might play an important role in promoting the growth of single crystals as opposed to polycrystals, in limiting nucleation, and in controlling the supersaturation of solution. The size, quality and growth rate of crystals might be controlled by adjusting content of ionic liquid.
离子液体具有独特的结构和物理化学性质,已经成为目前化学界的热点课题之一,而且在生命科学领域也开始得到应用,尤其是作为生物催化反应的溶剂或共溶剂的研究更是备受关注,近年来,离子液体也开始应用于蛋白质结晶方面。
本论文将一种新体系——离子液体-水用于蛋白质的结晶。在该体系中,实现了蛋白质晶体在离子液体中的可控生长,培养了符合X射线衍射条件的蛋白质单晶,对离子液体在蛋白质结晶过程中的作用机理进行了探讨,并剖析了离子液体含量与蛋白质晶体的晶型、形貌和品质间的关系。此外,从动力学角度系统地分析了离子液体在蛋白质结晶过程中对晶体生长速率的控制作用,进而优化结晶条件,改善晶体质量。
采用一步法和两步法分别合成了离子液体[BMIM]Cl(1-丁基-3-甲基咪唑氯化物)、[BMIM]BF_4(1-丁基-3-甲基咪唑四氟硼酸盐)和[BMIM]PF_6(1-丁基-3-甲基咪唑六氟磷酸盐),借助红外光谱分析表征了其化学结构。其次,为了进一步研究离子液体对蛋白质晶体生长过程的影响机制,采用BCA法通过紫外—可见分光光度计测定蛋白质在离子液体—水混合溶液中的溶解度,结果表明,离子液体在体系中起到了类似无机盐类沉淀剂的作用,促进了蛋白质分子从溶液中的析出。通过测定[BMIM]BF_4-乙酸乙酸钠-NaCl离子液体双水相体系的三元体系相图,研究了培养溶液中各组分间的溶解度关系,并将其用于蛋白质晶体的培养,以控制了溶液的局部过饱和度。
采用悬滴法和批量法两种结晶方法,在离子液体-水体系中实现了对溶菌酶晶体的可控生长,得到了空间群为P4_32_12的四方晶系晶体,并对其结构进行了模型解析工作。考察了沉淀剂种类、溶液过饱和度、离子液体的含量、结晶温度等实验参数的改变对溶菌酶晶型、晶体形貌和品质的影响。研究表明,[BMIM]BF_4更适于溶菌酶晶体的培养,在该体系中,[BMIM]BF_4与NaCl一起作为联合沉淀剂使用,晶体尺寸明显增大,结晶速率得以控制,晶体质量有所改善。另外,随着[BMIM]BF_4含量的增加,晶体形貌逐渐由块状变为棒状,且长径比也随之增大。
采用悬滴结晶法,在[BMIM]BF_4-水体系中实现了对索马甜蛋白单晶体的可控生长,得到了空间群为P4_12_12的四方晶系晶体,并解析了其初始模型结构。在该体系中,[BMIM]BF_4被单独的作为沉淀剂使用,并且研究了结晶温度、离子液体[BMIM]BF_4的含量等培养条件对索马甜蛋白晶型和品质的影响。
离子液体作为一种具有特殊结构和功能的溶剂,在蛋白质结晶的过程中具有与无机盐类沉淀剂相类似的作用。同时形成双水相体系降低溶液的局部过饱和度,发挥了模板剂、联合沉淀剂和添加剂的多重作用,有效的控制晶核的生成数量,提高单晶出现的几率。可以通过调节离子液体的含量来控制蛋白质晶体的生长速率、大小和质量。
Protein crystallography is an interdisciplinary subject in the research of the structure and function of the protein. With the increasing demands of precise biomolecular 3-D structures, especially protein structures by X-ray diffraction, people pay more and more attention to the study of protein crystal growth. However, the crystallization of protein is very difficult and become a bottleneck in structural genomics, to some extent.
Ionic liquids (ILs) with unique structure and outstanding physical and chemical properties have become the focus in the chemistry field. And it is also used in the field of life science, especially in the biocatalyst reaction as solvent or cosolvent. Recently, ionic liquids are also applied in protein crystallization.
In this work, a new system of ionic liquid-water was tested for the protein crystallization. The procedure of crystallization could be controllable conducted in this system. The mechanisms for formations of controlled growth and the relationship among polymorph, morphology, quality and ionic liquid content were investigated. Moreover, the limitation effect of ionic liquid on the rate of crystal growth was disscussed from the view of kinetics.
Ionic liquids of [BMIM]Cl, [BMIM]BF_4 and [BMIM]PF_6 were prepared through Direct Method and Two-step Method, respectively. And the chemical structures of the final products were characterized by Infrared spectra. For the further studies on the effects of the ILs on the crystal growth, the solubility of protein was mensurated with ultraviolet-visible light spectrophotometer by the way of BCA method. It was found that ionic liquid had similar effect as precipitants or additions, which was favourable for the single crystals. On the other hand, in order to understand their dissolved relationship among each components in the solution, the equilibrium solubility of a ternary system of [BMIM]BF_4, NaCl and buffer as well as the data of liquid-liquid equilibrium of that system are determined at 25℃and atmospheric pressure. It was used for the protein crystallization, and played a role in the conrol of local supersaturation.
Two crystallization methods (hanging drop and sitting drop crystallization) were used for the lysozyme crystallization in the ionic liquids with different buffer/IL proportions. Lysozyme crystal belonged to the tetragonal space group P4_32_12 and its 3-D model was analyzed. The influence of growth conditions such as the kind of precipitants, solution supersaturation, ionic liquids content and temperature on polymorph, morphology, quality of crystals was discussed. It showed that [BMIM]BF_4 was more suitable for lysozyme crystallization than other ILs. [BMIM]BF_4 and NaCl were used as union-precipitants in the system of [BMIM]BF_4-water. More lager single crystals with controllable morphologies could be obtained due to the manageable crystal growth velocity. In addition, the morphology of lysozyme crystals were changed from bolck to rod and the L/D ratio increased with the proportion of IL.
Futhermore, thaumatin crystal, which was belonged to the tetragonal space group P4_12_12, could be obtained with hanging drop crystallization. [BMIM]BF_4 was used as a precipitant, and the effect of ionic liquids content, temperature on polymorph and quality of crystals was investigated.
On a whole, ionic liquid was a solvent with special structure and function. More importantly, aqueous two-phase system was appeared with diffusion of water molecules. As a new system, it might play an important role in promoting the growth of single crystals as opposed to polycrystals, in limiting nucleation, and in controlling the supersaturation of solution. The size, quality and growth rate of crystals might be controlled by adjusting content of ionic liquid.
引文
[1]Mark B,Haitao Y and Zihe R.Drug Design Targeting the Main Protease,the Achilles' Heel of Coronaviruses[J].Curr Opin Struct Biol,2005,15:664-672P
[2]Sun F,Huo X,Zhai Y J,et al.Crystal structure of mitochondrial respiratory membrane protein complex Ⅱ[J].Cell,2005,121:1043-1057P
[3]舒占永,毕汝昌.蛋白质晶体的优化生长[J].生物化学与生物物理进展,1997,24(5):396-401页
[4]马润宇,王艳辉,涂感良.膜结晶技术研究进展及应用前景[J].膜科学与技术,2003,23(4):145-150页
[5]桂林,李林,胡松青等.生物大分子结晶的研究进展[J].化工进展,2005,24(8):854-859页
[6]卢光莹,华子千.生物大分子晶体学基础[M].北京:北京大学出版社,2001:5-6页
[7]McPherson A.Crystallization of biological macromolecules[M].New York:Cold Spring Harbor,1999:173-176P
[8]王弘,于淑娟,高大维.蛋白质大分子的结晶[J].生命的化学,2001,21(5):429-432页
[9]何忠效.生物化学实验技术[M].北京:化学工业出版社,2004:327-331页
[10]Mcpherson A.Use of Polyethylene Glycol in the Crystallizaition of Mactomolecules[J].Meth.Enzymol.,1985b,114:120-125P
[11]Markman O,Rob C,Roberts M F,et al.Designed additives for controlled growth of crystals of phospholipids interacting proteins:Short chain phospholipids[J].J Cryst Growth,1996,160:382-388P
[12]Tanaka S,Ataka M,Kubota T,et al.The effect of amphiphilic additives on the growth and morphology of Aspergillus niger acidproteinase A crystals[J].J Cryst Growth,2002,234:247-254P
[13]Lu J,Wang X J and Ching C B.Batch crystallization of soluble proteins:Effect of precipitant,temperature and additive[J].Prog Cryst Growth Charact Mater,2002:201-217P
[14]McPherson A,Malkin A J and Kuznetsov Y G.The Science of Macromoleular Crystallization[J].Structure,1995,3:759-768P
[15]DeLucas L J,Smith C D,Smith H W,et al.Protein crystal growth in microgravity[J].Science,1989,246(4930):651-654P
[16]McPherson A.Recent Advances in the Microgravity Crystallization of Biological Macromolecules[J].J Trends Biotechnol,1997,15(6):197-237P
[17]Penkova A,Gliko O,Dimitrov I L,et al.Enhancement and suppression of protein crystal nucleation due to electrically driven convection[J].J Cryst Growth,2005,275:1527-1532P
[18]Motokaw M,Watanabe K and Awaji S.High magnetic field researches in Tohoku University[J].Curr Appl Phys,2003,3(4):367-376P
[19]Kakinouchi K,Adachib H,Matsumura H,et al.Effect of ultrasonic irradiation on protein crystallization[.T].J Cryst Growth,2006,292:437-440P
[20]Kitano H,Murakamic S,Adachi H,et al.Processing of membrane protein crystal using ultraviolet laser irradiation[J].J Biosci Bioeng,2005,100(1):50-53P
[21]McPherson A.Macromolecular crystals[J].Sci Am,1989,260:62-69P
[22]Matthews B W.Solvent content of protein crystals[J].J Mol Biol,1968,33:491-497P
[23]McPherson A.Crystallization of biological macromolecules[M].New York:Cold Spring Harbor,1999:148-150P
[24]Koch V R,Miller L L and Osteryoung R A.Electroinitiated Friede 1-Crafts Transalky-lations in a Room Temperature Molten-Salt Medium[J].J Am Chem Soc,1976,98(18):5277-5284P
[25]Kubisa P.Application of ionic liquids as solvents for polymerization processes[J].Polym Prog.Sci,2004,29:3-12P
[26]Dzyuba S V and Bartsch R A.Recent advances in applications of room-temperature ionic liquid/supercritical CO_2 systems[J].Angew Chem Int Ed,2003,42:148-150P
[27]Huddleston J G,Willauer H D,Swatloski R P,et al.Room Temperature Ionic Liquids as Novel Media for 'Clean' Liquid-Liquid Extraction[J].Chem Commun,1998,24:1765-1766P
[28]Dupont J,Suoza R F,Suarez P A Z,et al.Ionic Liquid(Molten Salt) Phase Organometallic Catalysis[J].Chem Rev,2002,102:3667-3692P
[29]Sheldon R.Catalytic reactions in ionic liquids[J].Chem Commun,2001,23:2399-2407P
[30]Endres F and Abedin S Z E.Electrodeposition of stable and narrowly dispersed germanium nanoclusters from an ionic liquid[J].Chem Commun,2002,28:892-893P
[31]Fuller J,Carkin R T,Osteryoung R A,et al.The room temperature ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate:Electrochemical couples and physical properties[J].J Electrochem Soc,1997,144:3881-3886P
[32]邓友全.离子液体-性质、制备与应用[M].北京:中国石化出版社,2006:1-98页
[33]王均凤,张锁江,陈慧萍等.离子液体的性质及其在催化反应中的应用[J].过程工程学报,2003,3(2):177-185P
[34]张锁江,吕兴梅.离子液体——从基础研究到工业应用[M].北京:科 学出版社,2006:60-62页
[35]Wilkes J S,Levisky J A,Wilson R A,et al.Dialkylimidazolium chloroaluminate melts:A new class of room-temperature ionic liquids for electrochemistry,spectroscopy,and synthesis[J].Inorg Chem,1982,21(3):1263-1264P
[36]王军,杨许召,吴诗德等.离子液体的性能及应用[M].北京:中国纺织出版社,2007:131-132页
[37]Fannin Jr A A,Floreani D A,King L A,et al.Properties of 1,3-dialkylimidazolium chloride-aluminum chloride ionic liquids.2.Phase transitions,densities,electrical conductivities,and viscosities[J].J Phys Chem,1984,88:2614-2621P
[38]Sun J,Forsyth M and Mac-Farlane D R.Room-Temperature Molten Salts Based on the Quaternary Ammonium Ion[J].J Phys Chem,1998,102:8858-8864P
[39]Perry L,Jones K M,Scott W D,et al.Densities,viscosities,and conductivities of mixtures of selected organic cosolvents with the Lewis basic aluminum-chloride plus 1-methyl-3-ethylimidazolium chloride molten salt[J].J Chem Eng Data,1995,40:615-619P
[40]Dzyuba S V and Bartsch R A.Influence of structural variations in 1-alkyl(aralkyl)-3-methylimidazolium hexafluorophosphates and bis(tri-fluoromethylsulfonyl)imides on physical properties of the ionic liquids[J].Chem Phys Chem,2002,3(2):161-166P
[41]Bonh(?)te P,Dias A P,Papageorigou N et al.Hydrophobic,highly conductive ambient-temperature molten salts[J].Inorg Chem,1996,35(5):1168-1178P
[42]Ngo H L,LeCompte K,Hargens L et al.Thermal properties of imidazolium ionic liquids[J].Thermochim Acta,2000,357:97-102P
[43]Huddleston J G,Visser A E,Reichert W M,et al.Characterization and comparison of hydrophilic and hydrophobic room temperature ionic liquids incorporating the imidazolium cation[J].Green Chem,2001,3(4):156-164P
[44]Nishida T,Tashiro Y and Yamamoto M.Physical and electrochemical properities of 1-alkyl-3-methy-limidazolium tetrafluoroborate for electrolyte[J].J Fluorine Chem,2003,120(2):135-141P
[45]Cole A C,Jensen J L,Ntai I,et al.Novel Bronsted Acidic Ionic Liquids and Their Use as Dual Solvent-Catalysts[M].J Am Chem Soc,2002:5692-5693P
[46]Ann E V,Reichert W M,Swatloski R P,et al.Characterization of hydrophilic and hydrophobic ionic liquids:Alternatives to volatile organic compounds for liquid-liquid separations[A].ACS Symp Ser[C].Washington,D.C.:American Chemical Society,2002:289-308P
[47]Karodia N,Guise S,Newlands C,et al.Clean catalysis with ionic solvents-phosphonium tosylates for hydroformylation[J].Chem Comm,1998,2341-2342P
[48]Wasserscheid P and Keim W.Ionic liquids-new solutions for transition metal catalysis[J].Angew Chem Int Ed,2000,39:3772-3789P
[49]钟平.室温离子液体的合成及其在有机合成中的应用[J].温州师范学院学报,2004,25(2):1-8页
[50]李汝雄,王建基.离子液体的热物理性质研究进展[J].北京石油化工学院学报,2005,13(3):20-23页
[51]Zaks A and Klibanov A M.Enzymatic catalysis in organic media at 100℃[J].Science,1984,224:1249-1251P
[52]Jiang T F,Gu Y L,Liang B et al.Dynamically coating the capillary with 1-alkyl-3-methylimidazolium-based ionic liquids for separation of basic proteins by capillary electrophoresis[J].Anal Chim Acta,2003,479: 249-254P
[53]Turner M B,Holbrey J D,Spear S K,et al.In Ionic Liquids ⅢB:Fundamentals,Progress,Challenges,and Opportunities[A].ACS Symp Ser[C].Washington,D.C.:American Chemical Society,2004:33-48P
[54]Garlitz J A,Summers C A,Flowers R A et al.Borgstah Ethylammonium nitrate:a protein crystallization reagent[J].Acta Cryst,1999,D55:2037-2038P
[55]Pusey M L,Paley M S,Turner M B,et al.Protein Crystallization Using Room Temperature Ionic Liquids[J].Cryst Growth Des,2007,7(4):787-793P
[56]王大成.蛋白质工程[M].北京:化学工业出版社,2005:171-175页
[57]Huddleston J G.,Visser A E,Reichert W M,et al.Characterization and comparison of hydrophilic and hydrophobic room temperature ionic liquids incorporating the imidazolium cation[J].Green Chemistry,2001,3:156-164P
[58]Lin J Q,Tan P H,Jin C Y,et al.Measurement and correalation of equilibrium data for aqueous two-phase system ethanol+ water+K_2HPO_4[J].Chem Res Chinese U,2004,20(2):195-199P
[59]李占双,景晓燕.现代分析测试技术[M].哈尔滨:哈尔滨工程大学出版社,2005:259-268页
[60]Wetlaufer D B.Ulatratiolet Spectra of Proteins and Amino Acids[J].Adv.Prot.Chem.,1962,17:303-309P
[61]Akins R F.Measurement of Protein in 20 seconds using a Microware BCA Assay[J].BioTechniques,1992,12:496-499P
[62]Jiang E Y,Palmer R A,Nancy E B,et al.Phase-Resolved Depth Profiling of Thin-Layered Plasma Polymer Films by Step-Scan Fourier Transform Infrared Photoacoustic Spectroscopy[J].Appl.Spectrosc.,1997,51: 1238-1244P
[63]Palmer R A,Chao J L,Dittmar R M,et al.Investigation of Time-Dependent Phenomena by Use of Step-Scan FT-IR[J].Appl.Spectrosc.,1993,47:1297-1310P
[64]Heinrichs W,Heinnchs M,Schonert H.Growth of protein single crystals in a periodically changing solubility gradient[J].J Cryst Growth,1992,122:186-193P
[65]宁永成.有机化合物结构鉴定及有机波谱学[M].北京:科学出版社,2005:322-332页
[66]高金森,王鹏,董坤等.氯化烷基咪唑离子液体分子结构和红外光谱的模拟计算[J].石油学报,2006,22(1):72-76页
[67]Thomazeau C C,Olivier-Bourbigou H L,Magna L,et al.Determination of an acidic scale in room temperature ionic liquids[J].J Am Chem Soc,2003,125(18):5254-5265P
[68]Sergey I N,Claude B and Philippe M.Instability ofactinide(Ⅳ) hexachloro complexes in room-temperature ionic liquid[BuMeIm]PF_6 due to hydrolysis of the hexafluorophosphate anion[J].Comptes Rendus Chimie,2007,10:1122-1127P
[69]Li S H,He C Y,Liu H W,et al.Ionic liquid-based aqueous two-phase system,a sample pretreatment procedure prior to high-performance liquid chromatography of opium alkaloids[J].J Chromatogr B,2005,826:58-62P
[70]Gutowski E K,Broker A G,Rogers D R,et al.Controlling the aqueous miscibility of ionic liquids:aqueous biphasic systems of water-miscible ionic liquids and water-structuring salts for recycle,metathesis,and separations[J].J·Am·Chem·Soc·,2003,125:6632-6633P
[71]Liu S J.Liquid-liquid equilibrium for the new aqueous two-phase system of ionic liquids-salt-water and its application[J].Green Chem,2006,8: 639-646P
[72]Smith P K,Krohn R I,Hermanson G T,et al.Measurement of Protein Using Bicinchoninic Acid[J].Anal.Biochem.,1985,150:76-85P
[73]Mcpherson A.Crystallization of macromolecules:General principles[J].Method Enzymol,1985,14:112-120P
[74]吴晓英,林影,陈慧英.溶菌酶的研究进展[J].工业微生物,2002,32(4):55-58页
[75]Alderton G and Fevold H L.Direct crystallization of lysozyme from egg white and some crystalline salts of lysozyme[J].J.Bid.Chem,1946,164:1-5P
[76]董君,韩青,储乃明等.空间生长的溶菌酶晶体的结构研究[J].科学通报,1994,39(24):2264-2267页
[77]Welton T.Room-Temperature Ionic Liquids Solvents for Synthesis and Catalysis[J].Chem Re,1999,99(8):2071-2083P
[78]梁栋材.X射线晶体学基础[M].北京:科学出版社,2006:1-6页
[79]Cantor C R and Schimmel P R.Biophysical Chemistry[M].New York:W.H.Freeman and Company,1980:849-866P
[80]Blundell T L and Johnson L N.Protein Crystallography[M].San Diego:Academic Press Inc,1976:230-235P
[81]汪家政,范明.蛋白质技术手册[M].北京:科学出版社,2000:264-265页
[82]Mcpherson A.Crystallizaition of Mactomolecules:General Principles[J].Meth.Enzymol.,1985a,14:112-120P
[83]徐兴,毕汝昌.T4溶菌酶晶体分子堆积的研究[J].生物物理学报,2000,16(4):674-679页
[84]Bernstein F C,Koetzle T F,Willams G J B,et al.The Protein Data Bank:A computer-based archival file for macromolecular structures[J].J.Mol.Biol, 1977,112:535-542P
[85]温元凯,邵俊.离子极化导论[M].合肥:安徽教育出版社,1985:97-104页
[86]天津大学无机化学教研室.无机化学[M].北京:高等教育出版社,1997:284-286页
[87]Darby N J and Creighton T E.Protein Structure[M].New York:Oxford University Press,1993:4-5P
[88]Nanev C and Penkova A.Nucleation of lysozyme crystals under external electric field[J].J.Cryst.Growth,2001,232:285-293P
[89]Steinrauf L K.Preliminary X-ray data for some new crystalline forms of -lactoglobulin and hen-egg-white lysozyme[J].Acta Crystallogr,1959,12:77-79P
[90]Crick F H C.The unit cells of four proteins[J].Acta Crystallogr,1953,6:221-222P
[91]Pan X J and Glatz C E.Solvent effects on the growth kinetics of subtilisin crystals[J].J.Cryst.Growth,2002,240:549-559P
[92]Rosenberger F.Protein crystallization[J].J.Cryst.Growth,1996,166:40-54P
[93]Caraballo K G,Baird J K and Ng J D.Comparison of the crystallization kinetics of canavalin and lysozyme[J].Cryst.Growth Des,2006,6:874-880P
[94]Taleb M,Didierjean C,Jelsch C,et al.Equilibrium kinetics of lysozyme crystallization under an external electric field[J].J.Cryst.Growth,2001,232:250-255P
[95]Randolph A D and Larson M A.Theory of Particulate Processes[M].New York:Academic Press Inc,1988:249-273P
[96]Rosenberger F.Protein crystallization[J].Materials Science Forum,1998, 241:276-277P
[97]韩毅,仓怀兴,毕汝昌.NaCl溶液体系溶菌酶结晶相图的测定[J].生物物理学报,1998,14(3):573-576页
[98]Gernert K M,Smith R,Carter D C.A Simple Apparatus for Controlling Nucleation and Size in Protein Crystal Growth[J].Analytical Biochemistry,1988,168:141-147P
[99]Wilson L J and Suddath F L.Control of solvent evaporation in hen egg white lysozyme crystallization[J].J Cryst Growth,1992,116:414-420P
[100]陶凤云,张新妙,马润宇.蛋白质结晶过程中的影响因素[J].化学工艺与工程,2006,23(3):260-264页
[101]毕汝昌,桂璐璐,褛美珍.蛋白质的晶体生长及其进展[J].生物化学与生物物理进展,2002,18(4):394-398页
[102]代小虎,毕汝昌.沉淀剂类型对蛋白质结晶分子堆积的影响[J].生物物理学报,1990,17(4):246-250页
[103]陆杰,王静康.蛋白质沉淀结晶研究进展[J].化学工程,2001,29(6):52-56页
[104]Aradawa T and Timasheff S N.Theory of Protein Stability[J].Meth Enzymol,1985,114:49-77P
[105]Berchtold H,Reshetnikova L,Reiser C O A,et al.Crystal Structure of Active Elongation Factor Tu Reveals Major Comain Rearrangements[J].Nature,365:126-132P
[106]Kazushige O and Charles E G.Effects of pH and ionic strength on microfiltration of C.glutamicum[J].Journal of Membrane Science,1999,153:23-32P
[107]王廷华,邹晓莉.蛋白质理论与技术[M].北京:科学出版社,2005:38-39页
[108]Mannherz H G.Crystallization of Action in Complex with Actin-Bingding Proteins[J].J Biol Chem,1992,267:11661-11664P
[109]Sousa R.Use of Glycerol,Polyols and other Protein Struvture Stabilizing Agents in Protein Crystallization[J].Acta Cryst,1995,51:271-277P
[110]Kulkarni A M and Zukoski C F.Nanoparticle crystal nucleation:Influence of solution conditions[J].Langmuir,2002,18(8):3090-3099P
[111]Howard B.The solubility of hen egg-white lysozyme[J].J Cryst Growth,1988,90:94-104P
[112]Ewing F L,Forsythe E I,Woerdm,et al.Effects of purification on the crystallization of lysozyme[J].J Cryst Growth,1996,160:389-397P
[113]Patel S,Cudney B,and McPherson A.Polymeric Precipitants for the Crystallization of Macromolecules[J].Biochem.Biophys.Res.Comm.,1995,207:819-828P
[114]McPherson A.Preparation and Analysis of Protein Crystals[M].New York:Wiley Interscience,John Wiley and sons,1982:102-104P
[115]Sazaki G,Nagatoshi Y,Suzuki Y.Solubility of tetragonal and orthorhombic lysozyme crystals under high pressure[J].J.Crystal Growth,1999,196:204-209P
[116]Christopher G K,Phipps A G,Gray R J.Temperature-dependent solubility of selected proteins[J].J.Crystal Growth,1998,191:820-826P
[117]Bancel P A,Cajipe V B,Rodier F,et al.Laser seeding for biomolecular crystallization[J].J.Crystal Growth,1998,191:537-544P
[118]Drenth J and Haas J.Protein crystals and their stability[J].J.CrystalGrowth,1992,122:107-109P
[119]陆健.蛋白质纯化技术及应用[M].北京:化学工业出版社,2005:11-29页
[120]McPherson A.Crystallization of Biological Macromolecules[M].New York:Cold spring Harbor,1999:487-488P
[121]Baird J K,Hill S C,Clunie J C.Kinetics of protein crystal nucleation and growth in the batch method[J].J Crystal Growth,1999,196:220-225P
[122]Boistelle R,Astier J P.Crystallization.mechanisms in solution[J].J.CrystalGrowth,1988,90:14-30P
[123]Mikol V,Rodeau J L,Giege R.Experimental Determination of water equilibration rates in the hanging drop method of Protein crystallization[J].Analytical Biochemistry,1990,186:332-339P
[124]刘秋云,贺竹梅,曹俊等.利用农杆菌系统将超甜定基因导入烟草[J].遗传,1998,20(10):24-27页
[125]Lee J H,Weickmann J L,Koduri R K,et al.Expression of synthetic thaumatin genes in yeast[J].Biochcmistry,1988,27:5101-5107P
[126]杨成丽,刘德立.植物甜蛋白Thaumatin研究进展[J].武汉植物学研究2001,19(2):153-157页
[127]Charrona C,Robertb M-C,Capelleb B,et al.X-ray diffraction properties of protein crystals prepared in agarose gel under hydrostatic pressure[J].Journal of Crystal Growth,2002,245:321-333P
[128]Kuznetsov Y G.,Malkin A J,McPherson A.The influence of precipitant concentration on macromolecular crystal gowth mechanisms[J].Journal of Crystal Growth,2001,232:114-118P
[129]Juarez-Martinez G,Garza C,Castillo R,et al.A dynamic light scattering investigation of the nucleation and growth of thaumatin crystals[J].Crystal Growth,2001,232:119-131P
[130]Garcia-Ruiz J M,Moreno A.Growth kinetics of protein single crystals in the gel acupuncture technique[J].Journal of Crystal Growth,1997,178:393-401P
[131]Chayen N E.Crystallization with oils:a new dimension in macromolecular crystal growth[J].Journal of Crystal Growth,1999,196:434-441P
[132]Lopez-Jaramillo F J,Otalora F,Gavira J A.Protein crystal quality in diffusive environments and its evaluation [J].Journal of Crystal Growth,2003,247:177-184P
[133]Kadri A,.Tenner G.,Damakc M,et al.Crystallogenesis studies of proteins in agarose gel--combined effect of high hydrostatic pressure and pH[J].Journal of Crystal Growth,2003,257:390-402P
[134]Stanley K,David M,Joseph N J,et al.Protein crystal growth rates determined by time lapse microphotographyr[J].Journal of Crystal Growth,1991,110:177-181P
[2]Sun F,Huo X,Zhai Y J,et al.Crystal structure of mitochondrial respiratory membrane protein complex Ⅱ[J].Cell,2005,121:1043-1057P
[3]舒占永,毕汝昌.蛋白质晶体的优化生长[J].生物化学与生物物理进展,1997,24(5):396-401页
[4]马润宇,王艳辉,涂感良.膜结晶技术研究进展及应用前景[J].膜科学与技术,2003,23(4):145-150页
[5]桂林,李林,胡松青等.生物大分子结晶的研究进展[J].化工进展,2005,24(8):854-859页
[6]卢光莹,华子千.生物大分子晶体学基础[M].北京:北京大学出版社,2001:5-6页
[7]McPherson A.Crystallization of biological macromolecules[M].New York:Cold Spring Harbor,1999:173-176P
[8]王弘,于淑娟,高大维.蛋白质大分子的结晶[J].生命的化学,2001,21(5):429-432页
[9]何忠效.生物化学实验技术[M].北京:化学工业出版社,2004:327-331页
[10]Mcpherson A.Use of Polyethylene Glycol in the Crystallizaition of Mactomolecules[J].Meth.Enzymol.,1985b,114:120-125P
[11]Markman O,Rob C,Roberts M F,et al.Designed additives for controlled growth of crystals of phospholipids interacting proteins:Short chain phospholipids[J].J Cryst Growth,1996,160:382-388P
[12]Tanaka S,Ataka M,Kubota T,et al.The effect of amphiphilic additives on the growth and morphology of Aspergillus niger acidproteinase A crystals[J].J Cryst Growth,2002,234:247-254P
[13]Lu J,Wang X J and Ching C B.Batch crystallization of soluble proteins:Effect of precipitant,temperature and additive[J].Prog Cryst Growth Charact Mater,2002:201-217P
[14]McPherson A,Malkin A J and Kuznetsov Y G.The Science of Macromoleular Crystallization[J].Structure,1995,3:759-768P
[15]DeLucas L J,Smith C D,Smith H W,et al.Protein crystal growth in microgravity[J].Science,1989,246(4930):651-654P
[16]McPherson A.Recent Advances in the Microgravity Crystallization of Biological Macromolecules[J].J Trends Biotechnol,1997,15(6):197-237P
[17]Penkova A,Gliko O,Dimitrov I L,et al.Enhancement and suppression of protein crystal nucleation due to electrically driven convection[J].J Cryst Growth,2005,275:1527-1532P
[18]Motokaw M,Watanabe K and Awaji S.High magnetic field researches in Tohoku University[J].Curr Appl Phys,2003,3(4):367-376P
[19]Kakinouchi K,Adachib H,Matsumura H,et al.Effect of ultrasonic irradiation on protein crystallization[.T].J Cryst Growth,2006,292:437-440P
[20]Kitano H,Murakamic S,Adachi H,et al.Processing of membrane protein crystal using ultraviolet laser irradiation[J].J Biosci Bioeng,2005,100(1):50-53P
[21]McPherson A.Macromolecular crystals[J].Sci Am,1989,260:62-69P
[22]Matthews B W.Solvent content of protein crystals[J].J Mol Biol,1968,33:491-497P
[23]McPherson A.Crystallization of biological macromolecules[M].New York:Cold Spring Harbor,1999:148-150P
[24]Koch V R,Miller L L and Osteryoung R A.Electroinitiated Friede 1-Crafts Transalky-lations in a Room Temperature Molten-Salt Medium[J].J Am Chem Soc,1976,98(18):5277-5284P
[25]Kubisa P.Application of ionic liquids as solvents for polymerization processes[J].Polym Prog.Sci,2004,29:3-12P
[26]Dzyuba S V and Bartsch R A.Recent advances in applications of room-temperature ionic liquid/supercritical CO_2 systems[J].Angew Chem Int Ed,2003,42:148-150P
[27]Huddleston J G,Willauer H D,Swatloski R P,et al.Room Temperature Ionic Liquids as Novel Media for 'Clean' Liquid-Liquid Extraction[J].Chem Commun,1998,24:1765-1766P
[28]Dupont J,Suoza R F,Suarez P A Z,et al.Ionic Liquid(Molten Salt) Phase Organometallic Catalysis[J].Chem Rev,2002,102:3667-3692P
[29]Sheldon R.Catalytic reactions in ionic liquids[J].Chem Commun,2001,23:2399-2407P
[30]Endres F and Abedin S Z E.Electrodeposition of stable and narrowly dispersed germanium nanoclusters from an ionic liquid[J].Chem Commun,2002,28:892-893P
[31]Fuller J,Carkin R T,Osteryoung R A,et al.The room temperature ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate:Electrochemical couples and physical properties[J].J Electrochem Soc,1997,144:3881-3886P
[32]邓友全.离子液体-性质、制备与应用[M].北京:中国石化出版社,2006:1-98页
[33]王均凤,张锁江,陈慧萍等.离子液体的性质及其在催化反应中的应用[J].过程工程学报,2003,3(2):177-185P
[34]张锁江,吕兴梅.离子液体——从基础研究到工业应用[M].北京:科 学出版社,2006:60-62页
[35]Wilkes J S,Levisky J A,Wilson R A,et al.Dialkylimidazolium chloroaluminate melts:A new class of room-temperature ionic liquids for electrochemistry,spectroscopy,and synthesis[J].Inorg Chem,1982,21(3):1263-1264P
[36]王军,杨许召,吴诗德等.离子液体的性能及应用[M].北京:中国纺织出版社,2007:131-132页
[37]Fannin Jr A A,Floreani D A,King L A,et al.Properties of 1,3-dialkylimidazolium chloride-aluminum chloride ionic liquids.2.Phase transitions,densities,electrical conductivities,and viscosities[J].J Phys Chem,1984,88:2614-2621P
[38]Sun J,Forsyth M and Mac-Farlane D R.Room-Temperature Molten Salts Based on the Quaternary Ammonium Ion[J].J Phys Chem,1998,102:8858-8864P
[39]Perry L,Jones K M,Scott W D,et al.Densities,viscosities,and conductivities of mixtures of selected organic cosolvents with the Lewis basic aluminum-chloride plus 1-methyl-3-ethylimidazolium chloride molten salt[J].J Chem Eng Data,1995,40:615-619P
[40]Dzyuba S V and Bartsch R A.Influence of structural variations in 1-alkyl(aralkyl)-3-methylimidazolium hexafluorophosphates and bis(tri-fluoromethylsulfonyl)imides on physical properties of the ionic liquids[J].Chem Phys Chem,2002,3(2):161-166P
[41]Bonh(?)te P,Dias A P,Papageorigou N et al.Hydrophobic,highly conductive ambient-temperature molten salts[J].Inorg Chem,1996,35(5):1168-1178P
[42]Ngo H L,LeCompte K,Hargens L et al.Thermal properties of imidazolium ionic liquids[J].Thermochim Acta,2000,357:97-102P
[43]Huddleston J G,Visser A E,Reichert W M,et al.Characterization and comparison of hydrophilic and hydrophobic room temperature ionic liquids incorporating the imidazolium cation[J].Green Chem,2001,3(4):156-164P
[44]Nishida T,Tashiro Y and Yamamoto M.Physical and electrochemical properities of 1-alkyl-3-methy-limidazolium tetrafluoroborate for electrolyte[J].J Fluorine Chem,2003,120(2):135-141P
[45]Cole A C,Jensen J L,Ntai I,et al.Novel Bronsted Acidic Ionic Liquids and Their Use as Dual Solvent-Catalysts[M].J Am Chem Soc,2002:5692-5693P
[46]Ann E V,Reichert W M,Swatloski R P,et al.Characterization of hydrophilic and hydrophobic ionic liquids:Alternatives to volatile organic compounds for liquid-liquid separations[A].ACS Symp Ser[C].Washington,D.C.:American Chemical Society,2002:289-308P
[47]Karodia N,Guise S,Newlands C,et al.Clean catalysis with ionic solvents-phosphonium tosylates for hydroformylation[J].Chem Comm,1998,2341-2342P
[48]Wasserscheid P and Keim W.Ionic liquids-new solutions for transition metal catalysis[J].Angew Chem Int Ed,2000,39:3772-3789P
[49]钟平.室温离子液体的合成及其在有机合成中的应用[J].温州师范学院学报,2004,25(2):1-8页
[50]李汝雄,王建基.离子液体的热物理性质研究进展[J].北京石油化工学院学报,2005,13(3):20-23页
[51]Zaks A and Klibanov A M.Enzymatic catalysis in organic media at 100℃[J].Science,1984,224:1249-1251P
[52]Jiang T F,Gu Y L,Liang B et al.Dynamically coating the capillary with 1-alkyl-3-methylimidazolium-based ionic liquids for separation of basic proteins by capillary electrophoresis[J].Anal Chim Acta,2003,479: 249-254P
[53]Turner M B,Holbrey J D,Spear S K,et al.In Ionic Liquids ⅢB:Fundamentals,Progress,Challenges,and Opportunities[A].ACS Symp Ser[C].Washington,D.C.:American Chemical Society,2004:33-48P
[54]Garlitz J A,Summers C A,Flowers R A et al.Borgstah Ethylammonium nitrate:a protein crystallization reagent[J].Acta Cryst,1999,D55:2037-2038P
[55]Pusey M L,Paley M S,Turner M B,et al.Protein Crystallization Using Room Temperature Ionic Liquids[J].Cryst Growth Des,2007,7(4):787-793P
[56]王大成.蛋白质工程[M].北京:化学工业出版社,2005:171-175页
[57]Huddleston J G.,Visser A E,Reichert W M,et al.Characterization and comparison of hydrophilic and hydrophobic room temperature ionic liquids incorporating the imidazolium cation[J].Green Chemistry,2001,3:156-164P
[58]Lin J Q,Tan P H,Jin C Y,et al.Measurement and correalation of equilibrium data for aqueous two-phase system ethanol+ water+K_2HPO_4[J].Chem Res Chinese U,2004,20(2):195-199P
[59]李占双,景晓燕.现代分析测试技术[M].哈尔滨:哈尔滨工程大学出版社,2005:259-268页
[60]Wetlaufer D B.Ulatratiolet Spectra of Proteins and Amino Acids[J].Adv.Prot.Chem.,1962,17:303-309P
[61]Akins R F.Measurement of Protein in 20 seconds using a Microware BCA Assay[J].BioTechniques,1992,12:496-499P
[62]Jiang E Y,Palmer R A,Nancy E B,et al.Phase-Resolved Depth Profiling of Thin-Layered Plasma Polymer Films by Step-Scan Fourier Transform Infrared Photoacoustic Spectroscopy[J].Appl.Spectrosc.,1997,51: 1238-1244P
[63]Palmer R A,Chao J L,Dittmar R M,et al.Investigation of Time-Dependent Phenomena by Use of Step-Scan FT-IR[J].Appl.Spectrosc.,1993,47:1297-1310P
[64]Heinrichs W,Heinnchs M,Schonert H.Growth of protein single crystals in a periodically changing solubility gradient[J].J Cryst Growth,1992,122:186-193P
[65]宁永成.有机化合物结构鉴定及有机波谱学[M].北京:科学出版社,2005:322-332页
[66]高金森,王鹏,董坤等.氯化烷基咪唑离子液体分子结构和红外光谱的模拟计算[J].石油学报,2006,22(1):72-76页
[67]Thomazeau C C,Olivier-Bourbigou H L,Magna L,et al.Determination of an acidic scale in room temperature ionic liquids[J].J Am Chem Soc,2003,125(18):5254-5265P
[68]Sergey I N,Claude B and Philippe M.Instability ofactinide(Ⅳ) hexachloro complexes in room-temperature ionic liquid[BuMeIm]PF_6 due to hydrolysis of the hexafluorophosphate anion[J].Comptes Rendus Chimie,2007,10:1122-1127P
[69]Li S H,He C Y,Liu H W,et al.Ionic liquid-based aqueous two-phase system,a sample pretreatment procedure prior to high-performance liquid chromatography of opium alkaloids[J].J Chromatogr B,2005,826:58-62P
[70]Gutowski E K,Broker A G,Rogers D R,et al.Controlling the aqueous miscibility of ionic liquids:aqueous biphasic systems of water-miscible ionic liquids and water-structuring salts for recycle,metathesis,and separations[J].J·Am·Chem·Soc·,2003,125:6632-6633P
[71]Liu S J.Liquid-liquid equilibrium for the new aqueous two-phase system of ionic liquids-salt-water and its application[J].Green Chem,2006,8: 639-646P
[72]Smith P K,Krohn R I,Hermanson G T,et al.Measurement of Protein Using Bicinchoninic Acid[J].Anal.Biochem.,1985,150:76-85P
[73]Mcpherson A.Crystallization of macromolecules:General principles[J].Method Enzymol,1985,14:112-120P
[74]吴晓英,林影,陈慧英.溶菌酶的研究进展[J].工业微生物,2002,32(4):55-58页
[75]Alderton G and Fevold H L.Direct crystallization of lysozyme from egg white and some crystalline salts of lysozyme[J].J.Bid.Chem,1946,164:1-5P
[76]董君,韩青,储乃明等.空间生长的溶菌酶晶体的结构研究[J].科学通报,1994,39(24):2264-2267页
[77]Welton T.Room-Temperature Ionic Liquids Solvents for Synthesis and Catalysis[J].Chem Re,1999,99(8):2071-2083P
[78]梁栋材.X射线晶体学基础[M].北京:科学出版社,2006:1-6页
[79]Cantor C R and Schimmel P R.Biophysical Chemistry[M].New York:W.H.Freeman and Company,1980:849-866P
[80]Blundell T L and Johnson L N.Protein Crystallography[M].San Diego:Academic Press Inc,1976:230-235P
[81]汪家政,范明.蛋白质技术手册[M].北京:科学出版社,2000:264-265页
[82]Mcpherson A.Crystallizaition of Mactomolecules:General Principles[J].Meth.Enzymol.,1985a,14:112-120P
[83]徐兴,毕汝昌.T4溶菌酶晶体分子堆积的研究[J].生物物理学报,2000,16(4):674-679页
[84]Bernstein F C,Koetzle T F,Willams G J B,et al.The Protein Data Bank:A computer-based archival file for macromolecular structures[J].J.Mol.Biol, 1977,112:535-542P
[85]温元凯,邵俊.离子极化导论[M].合肥:安徽教育出版社,1985:97-104页
[86]天津大学无机化学教研室.无机化学[M].北京:高等教育出版社,1997:284-286页
[87]Darby N J and Creighton T E.Protein Structure[M].New York:Oxford University Press,1993:4-5P
[88]Nanev C and Penkova A.Nucleation of lysozyme crystals under external electric field[J].J.Cryst.Growth,2001,232:285-293P
[89]Steinrauf L K.Preliminary X-ray data for some new crystalline forms of -lactoglobulin and hen-egg-white lysozyme[J].Acta Crystallogr,1959,12:77-79P
[90]Crick F H C.The unit cells of four proteins[J].Acta Crystallogr,1953,6:221-222P
[91]Pan X J and Glatz C E.Solvent effects on the growth kinetics of subtilisin crystals[J].J.Cryst.Growth,2002,240:549-559P
[92]Rosenberger F.Protein crystallization[J].J.Cryst.Growth,1996,166:40-54P
[93]Caraballo K G,Baird J K and Ng J D.Comparison of the crystallization kinetics of canavalin and lysozyme[J].Cryst.Growth Des,2006,6:874-880P
[94]Taleb M,Didierjean C,Jelsch C,et al.Equilibrium kinetics of lysozyme crystallization under an external electric field[J].J.Cryst.Growth,2001,232:250-255P
[95]Randolph A D and Larson M A.Theory of Particulate Processes[M].New York:Academic Press Inc,1988:249-273P
[96]Rosenberger F.Protein crystallization[J].Materials Science Forum,1998, 241:276-277P
[97]韩毅,仓怀兴,毕汝昌.NaCl溶液体系溶菌酶结晶相图的测定[J].生物物理学报,1998,14(3):573-576页
[98]Gernert K M,Smith R,Carter D C.A Simple Apparatus for Controlling Nucleation and Size in Protein Crystal Growth[J].Analytical Biochemistry,1988,168:141-147P
[99]Wilson L J and Suddath F L.Control of solvent evaporation in hen egg white lysozyme crystallization[J].J Cryst Growth,1992,116:414-420P
[100]陶凤云,张新妙,马润宇.蛋白质结晶过程中的影响因素[J].化学工艺与工程,2006,23(3):260-264页
[101]毕汝昌,桂璐璐,褛美珍.蛋白质的晶体生长及其进展[J].生物化学与生物物理进展,2002,18(4):394-398页
[102]代小虎,毕汝昌.沉淀剂类型对蛋白质结晶分子堆积的影响[J].生物物理学报,1990,17(4):246-250页
[103]陆杰,王静康.蛋白质沉淀结晶研究进展[J].化学工程,2001,29(6):52-56页
[104]Aradawa T and Timasheff S N.Theory of Protein Stability[J].Meth Enzymol,1985,114:49-77P
[105]Berchtold H,Reshetnikova L,Reiser C O A,et al.Crystal Structure of Active Elongation Factor Tu Reveals Major Comain Rearrangements[J].Nature,365:126-132P
[106]Kazushige O and Charles E G.Effects of pH and ionic strength on microfiltration of C.glutamicum[J].Journal of Membrane Science,1999,153:23-32P
[107]王廷华,邹晓莉.蛋白质理论与技术[M].北京:科学出版社,2005:38-39页
[108]Mannherz H G.Crystallization of Action in Complex with Actin-Bingding Proteins[J].J Biol Chem,1992,267:11661-11664P
[109]Sousa R.Use of Glycerol,Polyols and other Protein Struvture Stabilizing Agents in Protein Crystallization[J].Acta Cryst,1995,51:271-277P
[110]Kulkarni A M and Zukoski C F.Nanoparticle crystal nucleation:Influence of solution conditions[J].Langmuir,2002,18(8):3090-3099P
[111]Howard B.The solubility of hen egg-white lysozyme[J].J Cryst Growth,1988,90:94-104P
[112]Ewing F L,Forsythe E I,Woerdm,et al.Effects of purification on the crystallization of lysozyme[J].J Cryst Growth,1996,160:389-397P
[113]Patel S,Cudney B,and McPherson A.Polymeric Precipitants for the Crystallization of Macromolecules[J].Biochem.Biophys.Res.Comm.,1995,207:819-828P
[114]McPherson A.Preparation and Analysis of Protein Crystals[M].New York:Wiley Interscience,John Wiley and sons,1982:102-104P
[115]Sazaki G,Nagatoshi Y,Suzuki Y.Solubility of tetragonal and orthorhombic lysozyme crystals under high pressure[J].J.Crystal Growth,1999,196:204-209P
[116]Christopher G K,Phipps A G,Gray R J.Temperature-dependent solubility of selected proteins[J].J.Crystal Growth,1998,191:820-826P
[117]Bancel P A,Cajipe V B,Rodier F,et al.Laser seeding for biomolecular crystallization[J].J.Crystal Growth,1998,191:537-544P
[118]Drenth J and Haas J.Protein crystals and their stability[J].J.CrystalGrowth,1992,122:107-109P
[119]陆健.蛋白质纯化技术及应用[M].北京:化学工业出版社,2005:11-29页
[120]McPherson A.Crystallization of Biological Macromolecules[M].New York:Cold spring Harbor,1999:487-488P
[121]Baird J K,Hill S C,Clunie J C.Kinetics of protein crystal nucleation and growth in the batch method[J].J Crystal Growth,1999,196:220-225P
[122]Boistelle R,Astier J P.Crystallization.mechanisms in solution[J].J.CrystalGrowth,1988,90:14-30P
[123]Mikol V,Rodeau J L,Giege R.Experimental Determination of water equilibration rates in the hanging drop method of Protein crystallization[J].Analytical Biochemistry,1990,186:332-339P
[124]刘秋云,贺竹梅,曹俊等.利用农杆菌系统将超甜定基因导入烟草[J].遗传,1998,20(10):24-27页
[125]Lee J H,Weickmann J L,Koduri R K,et al.Expression of synthetic thaumatin genes in yeast[J].Biochcmistry,1988,27:5101-5107P
[126]杨成丽,刘德立.植物甜蛋白Thaumatin研究进展[J].武汉植物学研究2001,19(2):153-157页
[127]Charrona C,Robertb M-C,Capelleb B,et al.X-ray diffraction properties of protein crystals prepared in agarose gel under hydrostatic pressure[J].Journal of Crystal Growth,2002,245:321-333P
[128]Kuznetsov Y G.,Malkin A J,McPherson A.The influence of precipitant concentration on macromolecular crystal gowth mechanisms[J].Journal of Crystal Growth,2001,232:114-118P
[129]Juarez-Martinez G,Garza C,Castillo R,et al.A dynamic light scattering investigation of the nucleation and growth of thaumatin crystals[J].Crystal Growth,2001,232:119-131P
[130]Garcia-Ruiz J M,Moreno A.Growth kinetics of protein single crystals in the gel acupuncture technique[J].Journal of Crystal Growth,1997,178:393-401P
[131]Chayen N E.Crystallization with oils:a new dimension in macromolecular crystal growth[J].Journal of Crystal Growth,1999,196:434-441P
[132]Lopez-Jaramillo F J,Otalora F,Gavira J A.Protein crystal quality in diffusive environments and its evaluation [J].Journal of Crystal Growth,2003,247:177-184P
[133]Kadri A,.Tenner G.,Damakc M,et al.Crystallogenesis studies of proteins in agarose gel--combined effect of high hydrostatic pressure and pH[J].Journal of Crystal Growth,2003,257:390-402P
[134]Stanley K,David M,Joseph N J,et al.Protein crystal growth rates determined by time lapse microphotographyr[J].Journal of Crystal Growth,1991,110:177-181P
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