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Ionic liquids in the life sciences: Are ionic liquids useful for the manipulation of biomolecules?

机译：生命科学中的离子液体：离子液体对操纵生物分子有用吗？

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摘要

Biocatalysis in non-aqueous media has become increasingly important over the past decade as the demand for enantiopure and other high-value added products has dramatically risen. The use of ionic liquids (ILs) as non-traditional, non-aqueous solvents for biocatalytic reactions has recently become an area of interest as preliminary results have shown both increased enantioselectivities and reaction rates. These results coupled with the potential for new chemistries in these "green" solvents have served to further propel this field. However, fundamental studies probing protein stability as a result of an IL environment have been lacking. In this work, the effects of various ILs on protein stability and methods by which improved stability may be ascertained were investigated.; This work outlines the benefit of medium engineering on protein stability. Novel ILs containing hydroxyl functionality have been prepared using a clean, atom-efficient synthesis resulting in a more favorable microenvironment for the biocatalyst compared to traditional alkylimidazolium ILs. Application of these ILs to biocatalytic processes appears to be promising as they have been shown to provide effective stabilization of model proteins.; The use of ILs in well-established methods to achieve increased enzymatic activity, such as enzyme immobilization, has also been investigated. In doing so, a new method has been developed for introducing active biocatalysts into cellulosic matrices using a cellulose-in-IL dissolution and regeneration process. Additionally, cellulose composite materials have been prepared providing primary amine surface functionality enabling direct, one-step bioconjugation.; Lastly, the denaturation of Trichoderma reesei cellulase in 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) has been shown to result in its inability to refold into its active form. However, covalent modification of the cellulase with a polyethylene glycol (PEG) polymer and modification of the reaction solvent through exchange of anionic components has been shown to drastically increase enzymatic activity.; Fundamental principles of protein stabilization in non-aqueous solvents have been applied to IL systems to achieve sustained or increased enzymatic activity and stability. The use of ILs as solvents for biocatalytic processes holds significant promise demonstrating the coupling of increased enzymatic activity afforded by its non-aqueous nature with the reduction in the use of volatile organic solvents.
机译：在过去的十年中，随着对映纯和其他高附加值产品的需求急剧增加，在非水介质中进行生物催化变得越来越重要。离子液体（ILs）作为生物催化反应的非传统，非水溶剂的使用近来已成为人们关注的领域，因为初步结果表明对映选择性和反应速率均得到提高。这些结果以及这些“绿色”溶剂中新化学的潜力，进一步推动了该领域的发展。但是，缺乏针对IL环境导致的蛋白质稳定性的基础研究。在这项工作中，研究了各种IL对蛋白质稳定性的影响以及可以确定改善的稳定性的方法。这项工作概述了蛋白质工程对蛋白质稳定性的好处。与传统的烷基咪唑类ILs相比，使用清洁，原子高效的合成方法制备了具有羟基官能团的新型ILs，从而为生物催化剂提供了更有利的微环境。这些ILs在生物催化过程中的应用似乎很有希望，因为它们已经显示出可以有效地稳定模型蛋白。还已经研究了在成熟的方法中使用IL来实现增加的酶活性，例如酶的固定化。为此，已经开发了一种新方法，用于使用IL-in-filament溶解和再生过程将活性生物催化剂引入到纤维素基质中。另外，已经制备了纤维素复合材料，其提供伯胺表面功能性，使得能够直接进行一步生物缀合。最后，已经证明里氏木霉纤维素酶在1-丁基-3-甲基咪唑鎓氯化物（[C4mim] Cl）中的变性导致其无法重折叠成其活性形式。然而，已经证明纤维素酶与聚乙二醇（PEG）聚合物的共价修饰和通过阴离子组分的交换对反应溶剂的修饰极大地提高了酶活性。非水溶剂中蛋白质稳定化的基本原理已应用于IL系统，以实现持续或增加的酶活性和稳定性。将IL用作生物催化过程的溶剂具有重大前景，这证明了其非水性质所提供的增加的酶活性与减少挥发性有机溶剂的使用相结合。

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  • 作者

    Turner, Megan Beth.;

  • 作者单位

    The University of Alabama.;

  • 授予单位 The University of Alabama.;
  • 学科 Chemistry Biochemistry.; Chemistry General.
  • 学位 Ph.D.
  • 年度 2005
  • 页码 165 p.
  • 总页数 165
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;化学;
  • 关键词

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